2017年12月, タマサート大学 名誉公衆衛生学博士号　　　　　　　　　　　　　　　
喜田 宏

2017年11月, 文化功労者　　　　　　　　　　　　　　　
喜田 宏

2017年05月, 春の叙勲 瑞宝重光章　　　　　　　　　　　　　　　
喜田 宏

2016年10月, 北海道功労賞　　　　　　　　　　　　　　　
インフルエンザウイルスの生態解明とその疾病対策
喜田 宏

2011年11月, 平成23年度 農事功績表彰緑白綬有功賞　　　　　　　　　　　　　　　
for Achievement of Control of Avian Influenza
喜田 宏

2009年02月, 平成20年度 畜産大賞　　　　　　　　　　　　　　　
インフルエンザウイルスの生態解明とライブラリーの構築－高病原性鳥インフルエンザの診断と予防への応用―
喜田宏

2005年06月, 第95回 日本学士院賞　　　　　　　　　　　　　　　
インフルエンザ制圧のための基礎的研究 ―家禽、家畜およびヒトの新型インフルエンザウイルスの出現機構の解明と抗体によるウイルス感染性中和の分子的基盤の確立
喜田宏

2005年04月, 平成17年度 日本農学賞・読売農学賞　　　　　　　　　　　　　　　
インフルエンザウイルスの生態に関する研究
喜田宏

2004年11月, 第58回 北海道新聞文化賞　　　　　　　　　　　　　　　
鳥、動物とヒトインフルエンザウイルスの生態学的研究
喜田宏

2002年03月, 北海道科学技術賞　　　　　　　　　　　　　　　
新型インフルエンザウイルスの出現機序の解明と対策に関する研究
喜田宏

1982年04月, 日本獣医学会賞　　　　　　　　　　　　　　　
鳥類パラミクソウイルスの分類に関する研究
喜田宏

Hemagglutinin and neuraminidase of a non-pathogenic H7N7 avian influenza virus coevolved during the acquisition of intranasal pathogenicity in chickens
Takaya Ichikawa, Takahiro Hiono, Masatoshi Okamatsu, Junki Maruyama, Daiki Kobayashi, Keita Matsuno, Hiroshi Kida, Yoshihiro Sakoda
Archives of Virology, 2024年10月
英語, 研究論文（学術雑誌）

Increased expression of CD38 on endothelial cells in SARS-CoV-2 infection in cynomolgus macaque
Cong Thanh Nguyen, Misako Nakayama, Hirohito Ishigaki, Yoshinori Kitagawa, Akemi Kakino, Marumi Ohno, Masashi Shingai, Yasuhiko Suzuki, Tatsuya Sawamura, Hiroshi Kida, Yasushi Itoh
Virology, 110052, 110052, Elsevier BV, 2024年03月
研究論文（学術雑誌）

Immunogenicity and protective efficacy of a co-formulated two-in-one inactivated whole virus particle COVID-19/influenza vaccine.
Chimuka Handabile, Marumi Ohno, Toshiki Sekiya, Naoki Nomura, Tomomi Kawakita, Mamiko Kawahara, Masafumi Endo, Tomohiro Nishimura, Minako Okumura, Shinsuke Toba, Michihito Sasaki, Yasuko Orba, Brendon Y Chua, Louise C Rowntree, Thi H O Nguyen, Masashi Shingai, Akihiko Sato, Hirofumi Sawa, Kazumasa Ogasawara, Katherine Kedzierska, Hiroshi Kida
Scientific reports, 14, 1, 4204, 4204, 2024年02月20日, ［国際誌］
英語, 研究論文（学術雑誌）, Due to the synchronous circulation of seasonal influenza viruses and severe acute respiratory coronavirus 2 (SARS-CoV-2) which causes coronavirus disease 2019 (COVID-19), there is need for routine vaccination for both COVID-19 and influenza to reduce disease severity. Here, we prepared individual WPVs composed of formalin-inactivated SARS-CoV-2 WK 521 (Ancestral strain; Co WPV) or influenza virus [A/California/07/2009 (X-179A) (H1N1) pdm; Flu WPV] to produce a two-in-one Co/Flu WPV. Serum analysis from vaccinated mice revealed that a single dose of Co/Flu WPV induced antigen-specific neutralizing antibodies against both viruses, similar to those induced by either type of WPV alone. Following infection with either virus, mice vaccinated with Co/Flu WPV showed no weight loss, reduced pneumonia and viral titers in the lung, and lower gene expression of proinflammatory cytokines, as observed with individual WPV-vaccinated. Furthermore, a pentavalent vaccine (Co/qFlu WPV) comprising of Co WPV and quadrivalent influenza vaccine (qFlu WPV) was immunogenic and protected animals from severe COVID-19. These results suggest that a single dose of the two-in-one WPV provides efficient protection against SARS-CoV-2 and influenza virus infections with no evidence of vaccine interference in mice. We propose that concomitant vaccination with the two-in-one WPV can be useful for controlling both diseases.

Extraction of the CDRH3 sequence of the mouse antibody repertoire selected upon influenza virus infection by subtraction of the background antibody repertoire
Masashi Shingai, Sayaka Iida, Naoko Kawai, Mamiko Kawahara, Toshiki Sekiya, Marumi Ohno, Naoki Nomura, Chimuka Handabile, Tomomi Kawakita, Ryosuke Omori, Junya Yamagishi, Kaori Sano, Akira Ainai, Tadaki Suzuki, Kazuo Ohnishi, Kimihito Ito, Hiroshi Kida
Journal of Virology, American Society for Microbiology, 2024年02月07日
研究論文（学術雑誌）, As specific interactions between antigens and cell-surface antibodies trigger the proliferation of B-cell clones, the frequency of each antibody sequence in the samples reflects the size of each clonal population. Nevertheless, it is extremely difficult to extract antigen-specific antibody sequences from the comprehensive bulk antibody sequences obtained from blood samples due to repertoire bias influenced by exposure to dietary antigens and other infectious agents. This issue can be addressed by subtracting the background noise from the post-immunization or post-infection repertoire data. In the present study, we propose a method to quantify repertoire data from comprehensive repertoire data. This method allowed subtraction of the background repertoire, resulting in more accurate extraction of expanded antibody repertoires upon influenza virus infection. This accurate extraction of antigen- or infection-specific repertoire information is a useful tool for vaccine evaluation.

The impact of PA/I38 substitutions and PA polymorphisms on the susceptibility of zoonotic influenza A viruses to baloxavir.
Keiichi Taniguchi, Takeshi Noshi, Shinya Omoto, Akihiko Sato, Takao Shishido, Keita Matsuno, Masatoshi Okamatsu, Scott Krauss, Richard J Webby, Yoshihiro Sakoda, Hiroshi Kida
Archives of virology, 169, 2, 29, 29, 2024年01月12日, ［国際誌］
英語, 研究論文（学術雑誌）, Genetic reassortment of avian, swine, and human influenza A viruses (IAVs) poses potential pandemic risks. Surveillance is important for influenza pandemic preparedness, but the susceptibility of zoonotic IAVs to the cap-dependent endonuclease inhibitor baloxavir acid (BXA) has not been thoroughly researched. Although an amino acid substitution at position 38 in the polymerase acidic protein (PA/I38) in seasonal IAVs reduces BXA susceptibility, PA polymorphisms at position 38 are rarely seen in zoonotic IAVs. Here, we examined the impact of PA/I38 substitutions on the BXA susceptibility of recombinant A(H5N1) viruses. PA mutants that harbored I38T, F, and M were 48.2-, 24.0-, and 15.5-fold less susceptible, respectively, to BXA than wild-type A(H5N1) but were susceptible to the neuraminidase inhibitor oseltamivir acid and the RNA polymerase inhibitor favipiravir. PA mutants exhibited significantly impaired replicative fitness in Madin-Darby canine kidney cells at 24 h postinfection. In addition, in order to investigate new genetic markers for BXA susceptibility, we screened geographically and temporally distinct IAVs isolated worldwide from birds and pigs. The results showed that BXA exhibited antiviral activity against avian and swine viruses with similar levels to seasonal isolates. All viruses tested in the study lacked the PA/I38 substitution and were susceptible to BXA. Isolates harboring amino acid polymorphisms at positions 20, 24, and 37, which have been implicated in the binding of BXA to the PA endonuclease domain, were also susceptible to BXA. These results suggest that monitoring of the PA/I38 substitution in animal-derived influenza viruses is important for preparedness against zoonotic influenza virus outbreaks.

2-thiouridine is a broad-spectrum antiviral nucleoside analogue against positive-strand RNA viruses.
Kentaro Uemura, Haruaki Nobori, Akihiko Sato, Shinsuke Toba, Shinji Kusakabe, Michihito Sasaki, Koshiro Tabata, Keita Matsuno, Naoyoshi Maeda, Shiori Ito, Mayu Tanaka, Yuki Anraku, Shunsuke Kita, Mayumi Ishii, Kayoko Kanamitsu, Yasuko Orba, Yoshiharu Matsuura, William W Hall, Hirofumi Sawa, Hiroshi Kida, Akira Matsuda, Katsumi Maenaka
Proceedings of the National Academy of Sciences of the United States of America, 120, 42, e2304139120, 2023年10月17日, ［国際誌］
英語, 研究論文（学術雑誌）, Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infections are causing significant morbidity and mortality worldwide. Furthermore, over 1 million cases of newly emerging or re-emerging viral infections, specifically dengue virus (DENV), are known to occur annually. Because no virus-specific and fully effective treatments against these or many other viruses have been approved, there is an urgent need for novel, effective therapeutic agents. Here, we identified 2-thiouridine (s2U) as a broad-spectrum antiviral ribonucleoside analogue that exhibited antiviral activity against several positive-sense single-stranded RNA (ssRNA+) viruses, such as DENV, SARS-CoV-2, and its variants of concern, including the currently circulating Omicron subvariants. s2U inhibits RNA synthesis catalyzed by viral RNA-dependent RNA polymerase, thereby reducing viral RNA replication, which improved the survival rate of mice infected with DENV2 or SARS-CoV-2 in our animal models. Our findings demonstrate that s2U is a potential broad-spectrum antiviral agent not only against DENV and SARS-CoV-2 but other ssRNA+ viruses.

The elucidation of plasma lipidome profiles during severe influenza in a mouse model.
Marumi Ohno, Siddabasave Gowda B Gowda, Toshiki Sekiya, Naoki Nomura, Masashi Shingai, Shu-Ping Hui, Hiroshi Kida
Scientific reports, 13, 1, 14210, 14210, 2023年08月30日, ［国際誌］
英語, 研究論文（学術雑誌）, Although influenza virus infection has been shown to affect lipid metabolism, details remain unknown. Therefore, we elucidated the kinetic lipid profiles of mice infected with different doses of influenza virus A/Puerto Rico/8/34 (H1N1) (PR8) by measuring multiple lipid molecular species using untargeted lipidomic analysis. C57BL/6 male mice were intranasally infected with PR8 virus at 50 or 500 plaque-forming units to cause sublethal or lethal influenza, respectively. Plasma and tissue samples were collected at 1, 3, and 6 days post-infection (dpi), and comprehensive lipidomic analysis was performed using high-performance liquid chromatography-linear trap quadrupole-Orbitrap mass spectrometry, as well as gene expression analyses. The most prominent feature of the lipid profile in lethally infected mice was the elevated plasma concentrations of phosphatidylethanolamines (PEs) containing polyunsaturated fatty acid (PUFA) at 3 dpi. Furthermore, the facilitation of PUFA-containing phospholipid production in the lungs, but not in the liver, was suggested by gene expression and lipidomic analysis of tissue samples. Given the increased plasma or serum levels of PUFA-containing PEs in patients with other viral infections, especially in severe cases, the elevation of these phospholipids in circulation could be a biomarker of infection and the severity of infectious diseases.

MGIT-seq for the Identification of Nontuberculous Mycobacteria and Drug Resistance: a Prospective Study.
Kiyoharu Fukushima, Yuki Matsumoto, Takanori Matsuki, Haruko Saito, Daisuke Motooka, Sho Komukai, Eriko Fukui, June Yamuchi, Tadayoshi Nitta, Takayuki Niitsu, Yuko Abe, Hiroshi Nabeshima, Yasuharu Nagahama, Takuro Nii, Kazuyuki Tsujino, Keisuke Miki, Seigo Kitada, Atsushi Kumanogoh, Shizuo Akira, Shota Nakamura, Hiroshi Kida
Journal of clinical microbiology, 61, 4, e0162622, 2023年03月22日, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, Because nontuberculous mycobacterial pulmonary disease is a considerable health burden, a simple and clinically applicable analytical protocol enabling the identification of subspecies and drug-resistant disease is required to determine the treatment strategy. We aimed to develop a simplified workflow consisting only of direct sequencing of mycobacterial growth indicator tube cultures (MGIT-seq). In total, 138 patients were prospectively enrolled between April 2021 and May 2022, and culture-positive MGIT broths were subjected to sequencing using MinION, a portable next-generation sequencer. Sequence analysis was conducted to identify species using core genome multilocus sequence typing and to predict macrolide and amikacin (AMK) resistance based on previously reported mutations in rrl, rrs, and erm(41). The results were compared to clinical tests for species identification and drug susceptibility. A total of 116 patients with positive MGIT cultures were included in the analysis. MGIT-seq yielded 99.1% accuracy in species-level identification and identified 98 isolates (84.5%) at the subspecies level. Macrolide and AMK resistance were detected in 19.4% and 1.9% of Mycobacterium avium complex (MAC) and Mycobacterium abscessus isolates. The predicted macrolide and AMK resistance was consistent with the results of conventional drug susceptibility tests, with specificities of 97.6% and 100.0%, respectively. Direct MGIT-seq has achieved comprehensive identification and drug resistance detection of nontuberculous mycobacteria, which could be applicable to determine the treatment strategy by a single test in clinical practice.

Assessing the pyrogenicity of whole influenza virus particle vaccine in cynomolgus macaques.
Marumi Ohno, Masataka Sagata, Toshiki Sekiya, Naoki Nomura, Masashi Shingai, Masafumi Endo, Kazuhiko Kimachi, Saori Suzuki, Cong Thanh Nguyen, Misako Nakayama, Hirohito Ishigaki, Kazumasa Ogasawara, Yasushi Itoh, Yoichiro Kino, Hiroshi Kida
Vaccine, 41, 3, 787, 794, 2023年01月16日, ［国際誌］
英語, 研究論文（学術雑誌）, Among inactivated influenza vaccines, the whole virus particle vaccine (WPV) elicits superior priming responses to split virus vaccine (SV) in efficiently inducing humoral and cellular immunity. However, there is concern for undesired adverse events such as fever for WPV due to its potent immunogenicity. Therefore, this study investigated the febrile response induced by subcutaneous injection with quadrivalent inactivated influenza vaccines of good manufacturing grade for pharmaceutical or investigational products in cynomolgus macaques. Body temperature was increased by 1 °C-2 °C for 6-12 h after WPV administration at the first vaccination but not at the second shot, whereas SV did not affect body temperature at both points. Given the potent priming ability of WPV, WPV-induced fever may be attributed to immune responses that uniquely occur during priming. Since WPV-induced fever was blunted by pretreatment with indomethacin (a cyclooxygenase inhibitor), the febrile response by WPV is considered to depend on the increase in prostaglandins synthesized by cyclooxygenase. In addition, WPV, but not SV, induced the elevation of type I interferons and monocyte chemotactic protein 1 in the plasma; these factors may be responsible for pyrogenicity caused by WPV, as they can increase prostaglandins in the brain. Notably, sufficient antibody responses were acquired by half the amount of WPV without causing fever, suggesting that excessive immune responses to trigger the febrile response is not required for acquired immunity induction. Thus, we propose that WPV with a reduced antigen dose should be evaluated for potential clinical usage, especially in naïve populations.

Effects of chondroitin sulfate oligosaccharides on osteoclast differentiation of RAW264 cells, and myotube differentiation of C2C12 cells.
Hirofumi Uchiyama, Daisuke Muramatsu, Hideaki Higashi, Hiroshi Kida, Atsushi Iwai
PloS one, 18, 4, e0284343, 2023年, ［国際誌］
英語, 研究論文（学術雑誌）, Chondroitin sulfate (CS) is a glycosaminoglycan, and CS derived from various animal species is used in drugs and food supplements to alleviate arthralgia. The CS is a high molecular weight compound, and hydrolysis of CS by intestinal microbiota is thought to be required for absorption in mammalians. Chondroitin sulfate oligosaccharides (Oligo-CS) are produced by hydrolysis with subcritical water from CS isolated from a species of skate, Raja pulchra for the improvement of bioavailability. The present study conducted in vitro experiments using murine cell lines, to compare the biological activities of Oligo-CS and high molecular weight CS composed with the similar disaccharide isomer units of D-glucuronic acid and N-acetyl-D-glucosamine (CS-C). The results show that Oligo-CS inhibits osteoclast differentiation of RAW264 cells significantly at lower concentrations than in CS. The cell viability of a myoblast cell line, C2C12 cells, was increased when the cells were grown in a differentiated medium for myotubes with Oligo-CS, where there were no effects on the cell viability in CS. These results suggest that in vitro Oligo-CS exhibits stronger bioactivity than high-molecular weight CS.

Effects of heat degradation of betanin in red beetroot (Beta vulgaris L.) on biological activity and antioxidant capacity.
Daisuke Muramatsu, Hirofumi Uchiyama, Hideaki Higashi, Hiroshi Kida, Atsushi Iwai
PloS one, 18, 5, e0286255, 2023年, ［国際誌］
英語, 研究論文（学術雑誌）, Betanin is a red pigment of red beetroot (Beta vulgaris L.), providing the beneficial effects to maintain human health. Betanin is involved in the characteristic red color of red beetroot, and used as an edible dye. Betanin is known to be a highly unstable pigment, and water solutions of betanin are nearly fully degraded after heating at 99°C for 60 min in the experimental conditions of this study. The present study investigated the effects of red beetroot juice (RBJ) and betanin on immune cells, and found that stimulation with RBJ and betanin induces interleukin (IL)-1β, IL-8, and IL-10 mRNA in a human monocyte derived cell line, THP-1 cells. This mRNA induction after stimulation with RBJ and betanin was not significantly changed after heat treatment when attempting to induce degradation of the betanin. Following these results, the effects of heat degradation of betanin on the inhibition of lipopolysaccharide (LPS) induced nitric oxide (NO) production in RAW264 cells and the antioxidant capacity were investigated. The results showed that the inhibition activity of RBJ and betanin with the LPS induced NO production is not altered after heat degradation of betanin. In addition, the results of FRAP (ferric reducing antioxidant power) and DPPH (1,1-Diphenyl-2-picrylhydrazyl) assays indicate that a not inconsiderable degree of the antioxidant capacity of RBJ and betanin remained after heat degradation of betanin. These results suggest that it is important to consider the effects of degradation products of betanin in the evaluation of the beneficial effects of red beetroot on health.

Field-deployable multiplex detection method of SARS-CoV-2 and influenza virus using loop-mediated isothermal amplification and DNA chromatography.
Kyoko Hayashida, Alejandro Garcia, Lavel Chinyama Moonga, Tatsuki Sugi, Kodera Takuya, Mitsuo Kawase, Fumihiro Kodama, Atsushi Nagasaka, Nobuhisa Ishiguro, Ayato Takada, Masahiro Kajihara, Naganori Nao, Masashi Shingai, Hiroshi Kida, Yasuhiko Suzuki, William W Hall, Hirofumi Sawa, Junya Yamagishi
PloS one, 18, 5, e0285861, 2023年, ［国際誌］
英語, 研究論文（学術雑誌）, A novel multiplex loop-mediated isothermal amplification (LAMP) method combined with DNA chromatography was developed for the simultaneous detection of three important respiratory disease-causing viruses: severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), influenza A virus, and influenza B virus. Amplification was performed at a constant temperature, and a positive result was confirmed by a visible colored band. An in-house drying protocol with trehalose was used to prepare the dried format multiplex LAMP test. Using this dried multiplex LAMP test, the analytical sensitivity was determined to be 100 copies for each viral target and 100-1000 copies for the simultaneous detection of mixed targets. The multiplex LAMP system was validated using clinical COVID-19 specimens and compared with the real-time qRT-PCR method as a reference test. The determined sensitivity of the multiplex LAMP system for SARS-CoV-2 was 71% (95% CI: 0.62-0.79) for cycle threshold (Ct) ≤ 35 samples and 61% (95% CI: 0.53-0.69) for Ct ≤40 samples. The specificity was 99% (95%CI: 0.92-1.00) for Ct ≤35 samples and 100% (95%CI: 0.92-1.00) for the Ct ≤40 samples. The developed simple, rapid, low-cost, and laboratory-free multiplex LAMP system for the two major important respiratory viral diseases, COVID-19 and influenza, is a promising field-deployable diagnosis tool for the possible future 'twindemic, ' especially in resource-limited settings.

Next-generation proteomics of serum extracellular vesicles combined with single-cell RNA sequencing identifies MACROH2A1 associated with refractory COVID-19.
Takahiro Kawasaki, Yoshito Takeda, Ryuya Edahiro, Yuya Shirai, Mari Nogami-Itoh, Takanori Matsuki, Hiroshi Kida, Takatoshi Enomoto, Reina Hara, Yoshimi Noda, Yuichi Adachi, Takayuki Niitsu, Saori Amiya, Yuta Yamaguchi, Teruaki Murakami, Yasuhiro Kato, Takayoshi Morita, Hanako Yoshimura, Makoto Yamamoto, Daisuke Nakatsubo, Kotaro Miyake, Takayuki Shiroyama, Haruhiko Hirata, Jun Adachi, Yukinori Okada, Atsushi Kumanogoh
Inflammation and regeneration, 42, 1, 53, 53, 2022年11月30日, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, BACKGROUND: The coronavirus disease 2019 (COVID-19) pandemic is widespread; however, accurate predictors of refractory cases have not yet been established. Circulating extracellular vesicles, involved in many pathological processes, are ideal resources for biomarker exploration. METHODS: To identify potential serum biomarkers and examine the proteins associated with the pathogenesis of refractory COVID-19, we conducted high-coverage proteomics on serum extracellular vesicles collected from 12 patients with COVID-19 at different disease severity levels and 4 healthy controls. Furthermore, single-cell RNA sequencing of peripheral blood mononuclear cells collected from 10 patients with COVID-19 and 5 healthy controls was performed. RESULTS: Among the 3046 extracellular vesicle proteins that were identified, expression of MACROH2A1 was significantly elevated in refractory cases compared to non-refractory cases; moreover, its expression was increased according to disease severity. In single-cell RNA sequencing of peripheral blood mononuclear cells, the expression of MACROH2A1 was localized to monocytes and elevated in critical cases. Consistently, single-nucleus RNA sequencing of lung tissues revealed that MACROH2A1 was highly expressed in monocytes and macrophages and was significantly elevated in fatal COVID-19. Moreover, molecular network analysis showed that pathways such as "estrogen signaling pathway," "p160 steroid receptor coactivator (SRC) signaling pathway," and "transcriptional regulation by STAT" were enriched in the transcriptome of monocytes in the peripheral blood mononuclear cells and lungs, and they were also commonly enriched in extracellular vesicle proteomics. CONCLUSIONS: Our findings highlight that MACROH2A1 in extracellular vesicles is a potential biomarker of refractory COVID-19 and may reflect the pathogenesis of COVID-19 in monocytes.

Immunization with inactivated whole virus particle influenza virus vaccines improves the humoral response landscape in cynomolgus macaques.
Brendon Y Chua, Toshiki Sekiya, Marios Koutsakos, Naoki Nomura, Louise C Rowntree, Thi H O Nguyen, Hayley A McQuilten, Marumi Ohno, Yuki Ohara, Tomohiro Nishimura, Masafumi Endo, Yasushi Itoh, Jennifer R Habel, Kevin J Selva, Adam K Wheatley, Bruce D Wines, P Mark Hogarth, Stephen J Kent, Amy W Chung, David C Jackson, Lorena E Brown, Masashi Shingai, Katherine Kedzierska, Hiroshi Kida
PLoS pathogens, 18, 10, e1010891, 2022年10月, ［国際誌］
英語, 研究論文（学術雑誌）, Although antibody-inducing split virus vaccines (SV) are currently the most effective way to combat seasonal influenza, their efficacy can be modest, especially in immunologically-naïve individuals. We investigated immune responses towards inactivated whole influenza virus particle vaccine (WPV) formulations, predicated to be more immunogenic, in a non-human primate model, as an important step towards clinical testing in humans. Comprehensive analyses were used to capture 46 immune parameters to profile how WPV-induced responses differed to those elicited by antigenically-similar SV formulations. Naïve cynomolgus macaques vaccinated with either monovalent or quadrivalent WPV consistently induced stronger antibody responses and hemagglutination inhibition (HI) antibody titres against vaccine-matched viruses compared to SV formulations, while acute reactogenic effects were similar. Responses in WPV-primed animals were further increased by boosting with the same formulation, conversely to modest responses after priming and boosting with SV. 28-parameter multiplex bead array defined key antibody features and showed that while both WPV and SV induced elevated IgG responses against A/H1N1 nucleoprotein, only WPV increased IgG responses against A/H1N1 hemagglutinin (HA) and HA-Stem, and higher IgA responses to A/H1N1-HA after each vaccine dose. Antibodies to A/H1N1-HA and HA-Stem that could engage FcγR2a and FcγR3a were also present at higher levels after one dose of WPV compared to SV and remained elevated after the second dose. Furthermore, WPV-enhanced antibody responses were associated with higher frequencies of HA-specific B-cells and IFN-γ-producing CD4+ T-cell responses. Our data additionally demonstrate stronger boosting of HI titres by WPV following prior infection and support WPV administered as a priming dose irrespective of the follow up vaccine for the second dose. Our findings thus show that compared to SV vaccination, WPV-induced humoral responses are significantly increased in scope and magnitude, advocating WPV vaccination regimens for priming immunologically-naïve individuals and also in the event of a pandemic outbreak.

Identification of cap-dependent endonuclease inhibitors with broad-spectrum activity against bunyaviruses.
Shinsuke Toba, Akihiko Sato, Makoto Kawai, Yoshiyuki Taoda, Yuto Unoh, Shinji Kusakabe, Haruaki Nobori, Shota Uehara, Kentaro Uemura, Keiichi Taniguchi, Masanori Kobayashi, Takeshi Noshi, Ryu Yoshida, Akira Naito, Takao Shishido, Junki Maruyama, Slobodan Paessler, Michael J Carr, William W Hall, Kumiko Yoshimatsu, Jiro Arikawa, Keita Matsuno, Yoshihiro Sakoda, Michihito Sasaki, Yasuko Orba, Hirofumi Sawa, Hiroshi Kida
Proceedings of the National Academy of Sciences of the United States of America, 119, 36, e2206104119, 2022年09月06日, ［国際誌］
英語, 研究論文（学術雑誌）, Viral hemorrhagic fevers caused by members of the order Bunyavirales comprise endemic and emerging human infections that are significant public health concerns. Despite the disease severity, there are few therapeutic options available, and therefore effective antiviral drugs are urgently needed to reduce disease burdens. Bunyaviruses, like influenza viruses (IFVs), possess a cap-dependent endonuclease (CEN) that mediates the critical cap-snatching step of viral RNA transcription. We screened compounds from our CEN inhibitor (CENi) library and identified specific structural compounds that are 100 to 1,000 times more active in vitro than ribavirin against bunyaviruses, including Lassa virus, lymphocytic choriomeningitis virus (LCMV), and Junin virus. To investigate their inhibitory mechanism of action, drug-resistant viruses were selected in culture. Whole-genome sequencing revealed that amino acid substitutions in the CEN region of drug-resistant viruses were located in similar positions as those of the CEN α3-helix loop of IFVs derived under drug selection. Thus, our studies suggest that CENi compounds inhibit both bunyavirus and IFV replication in a mechanistically similar manner. Structural analysis revealed that the side chain of the carboxyl group at the seventh position of the main structure of the compound was essential for the high antiviral activity against bunyaviruses. In LCMV-infected mice, the compounds significantly decreased blood viral load, suppressed symptoms such as thrombocytopenia and hepatic dysfunction, and improved survival rates. These data suggest a potential broad-spectrum clinical utility of CENis for the treatment of both severe influenza and hemorrhagic diseases caused by bunyaviruses.

Chronic Pulmonary Disease Caused by Tsukamurella toyonakaense.
Tomoki Kuge, Kiyoharu Fukushima, Yuki Matsumoto, Haruko Saito, Yuko Abe, Eri Akiba, Kako Haduki, Tadayoshi Nitta, Akira Kawano, Michio Tanaka, Yumi Hattori, Takahiro Kawasaki, Takanori Matsuki, Takayuki Shiroyama, Daisuke Motooka, Kazuyuki Tsujino, Keisuke Miki, Masahide Mori, Seigo Kitada, Shota Nakamura, Tetsuya Iida, Atsushi Kumanogoh, Hiroshi Kida
Emerging infectious diseases, 28, 7, 1437, 1441, 2022年07月, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, Unidentified Mycobacterium species are sometimes detected in respiratory specimens. We identified a novel Tsukamurella species (Tsukamurella sp. TY48, RIMD 2001001, CIP 111916T), Tsukamurella toyonakaense, from a patient given a misdiagnosis of nontuberculous mycobacterial pulmonary disease caused by unidentified mycobacteria. Genomic identification of this Tsukamurella species helped clarify its clinical characteristics and epidemiology.

Inactivated whole influenza virus particle vaccines induce neutralizing antibodies with an increase in immunoglobulin gene subclones of B-lymphocytes in cynomolgus macaques.
Masanori Shiohara, Saori Suzuki, Shintaro Shichinohe, Hirohito Ishigaki, Misako Nakayama, Naoki Nomura, Masashi Shingai, Toshiki Sekiya, Marumi Ohno, Sayaka Iida, Naoko Kawai, Mamiko Kawahara, Junya Yamagishi, Kimihito Ito, Ryotarou Mitsumata, Tomio Ikeda, Kenji Motokawa, Tomoyoshi Sobue, Hiroshi Kida, Kazumasa Ogasawara, Yasushi Itoh
Vaccine, 40, 30, 4026, 4037, 2022年06月26日, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, The All-Japan Influenza Vaccine Study Group has been developing a more effective vaccine than the current split vaccines for seasonal influenza virus infection. In the present study, the efficacy of formalin- and/or β-propiolactone-inactivated whole virus particle vaccines for seasonal influenza was compared to that of the current ether-treated split vaccines in a nonhuman primate model. The monovalent whole virus particle vaccines or split vaccines of influenza A virus (H1N1) and influenza B virus (Victoria lineage) were injected subcutaneously into naïve cynomolgus macaques twice. The whole virus particle vaccines induced higher titers of neutralizing antibodies against H1N1 influenza A virus and influenza B virus in the plasma of macaques than did the split vaccines. At challenge with H1N1 influenza A virus or influenza B virus, the virus titers in nasal swabs and the increases in body temperatures were lower in the macaques immunized with the whole virus particle vaccine than in those immunized with the split vaccine. Repertoire analyses of immunoglobulin heavy chain genes demonstrated that the number of B-lymphocyte subclones was increased in macaques after the 1st vaccination with the whole virus particle vaccine, but not with the split vaccine, indicating that the whole virus particle vaccine induced the activation of vaccine antigen-specific B-lymphocytes more vigorously than did the split vaccine at priming. Thus, the present findings suggest that the superior antibody induction ability of the whole virus particle vaccine as compared to the split vaccine is attributable to its stimulatory properties on the subclonal differentiation of antigen-specific B-lymphocytes.

Longitudinal alterations of the gut mycobiota and microbiota on COVID-19 severity.
Yuichi Maeda, Daisuke Motooka, Takahiro Kawasaki, Hiroya Oki, Yoshimi Noda, Yuichi Adachi, Takayuki Niitsu, Shota Okamoto, Kentaro Tanaka, Kiyoharu Fukushima, Saori Amiya, Reina Hara, Eri Oguro-Igashira, Takanori Matsuki, Haruhiko Hirata, Yoshito Takeda, Hiroshi Kida, Atsushi Kumanogoh, Shota Nakamura, Kiyoshi Takeda
BMC infectious diseases, 22, 1, 572, 572, 2022年06月24日, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, BACKGROUND: The impact of SARS-CoV-2 infection on the gut fungal (mycobiota) and bacterial (microbiota) communities has been elucidated individually. This study analyzed both gut mycobiota and microbiota and their correlation in the COVID-19 patients with severe and mild conditions and follow-up to monitor their alterations after recovery. METHODS: We analyzed the gut mycobiota and microbiota by bacterial 16S and fungal ITS1 metagenomic sequencing of 40 severe patients, 38 mild patients, and 30 healthy individuals and reanalyzed those of 10 patients with severe COVID-19 approximately 6 months after discharge. RESULTS: The mycobiota of the severe and mild groups showed lower diversity than the healthy group, and in some, characteristic patterns dominated by a single fungal species, Candida albicans, were detected. Lower microbial diversity in the severe group was observed, but no differences in its diversity or community structure were detected between the mild and healthy groups. The microbiota of the severe group was characterized by an increase in Enterococcus and Lactobacillus, and a decrease in Faecalibacterium and Bacteroides. The abundance of Candida was positively correlated with that of Enterococcus in patients with COVID-19. After the recovery of severe patients, alteration of the microbiota remained, but the mycobiota recovered its diversity comparable to that of mild and healthy groups. CONCLUSION: In mild cases, the microbiota is stable during SARS-CoV-2 infection, but in severe cases, alterations persist for 6 months after recovery.

Inactivated Whole Virus Particle Influenza Vaccine Induces Anti-Neuraminidase Antibodies That May Contribute to Cross-Protection against Heterologous Virus Infection.
Chimuka Handabile, Toshiki Sekiya, Naoki Nomura, Marumi Ohno, Tomomi Kawakita, Masashi Shingai, Hiroshi Kida
Vaccines, 10, 5, 2022年05月19日, ［国際誌］
英語, 研究論文（学術雑誌）, Despite the use of vaccines, seasonal influenza remains a risk to public health. We previously proposed the inactivated whole virus particle vaccine (WPV) as an alternative to the widely used split vaccine (SV) for the control of seasonal and pandemic influenza based on the superior priming potency of WPV to that of SV. In this study, we further examined and compared the immunological potency of monovalent WPV and SV of A/California/7/2009 (X-179A) (H1N1) pdm09 (CA/09) to generate immune responses against heterologous viruses, A/Singapore/GP1908/2015 (IVR-180) (H1N1) pdm09 (SG/15), and A/duck/Hokkaido/Vac-3/2007 (H5N1) (DH/07) in mice. Following challenge with a lethal dose of heterologous SG/15, lower virus titer in the lungs and milder weight loss were observed in WPV-vaccinated mice than in SV-vaccinated ones. To investigate the factors responsible for the differences in the protective effect against SG/15, the sera of vaccinated mice were analyzed by hemagglutination-inhibition (HI) and neuraminidase-inhibition (NI) assays to evaluate the antibodies induced against viral hemagglutinin (HA) and neuraminidase (NA), respectively. While the two vaccines induced similar levels of HI antibodies against SG/15 after the second vaccination, only WPV-vaccinated mice induced significantly higher titers of NI antibodies against the strain. Furthermore, given the significant elevation of NI antibody titers against DH/07, an H5N1 avian influenza virus, WPV was also demonstrated to induce NA-inhibiting antibodies that recognize NA of divergent strains. This could be explained by the higher conservation of epitopes of NA among strains than for HA. Taking these findings together, NA-specific antibodies induced by WPV may have contributed to better protection from infection with heterologous influenza virus SG/15, compared with SV. The present results indicate that WPV is an effective vaccine for inducing antibodies against both HA and NA of heterologous viruses and may be a useful vaccine to conquer vaccine strain mismatch.

Leptospira Is an environmental bacterium that grows in waterlogged soil.
Yasutake Yanagihara, Sharon Y A M Villanueva, Naoki Nomura, Marumi Ohno, Toshiki Sekiya, Chimuka Handabile, Masashi Shingai, Hideaki Higashi, Shin-Ichi Yoshida, Toshiyuki Masuzawa, Nina G Gloriani, Mitsumasa Saito, Hiroshi Kida
Microbiology Spectrum, 10, 2, e0215721, 2022年03月15日, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, Leptospirosis is a zoonotic disease caused by infection with pathogenic leptospires. Consistent with recent studies by other groups, leptospires were isolated from 89 out of 110 (80.9%) soil or water samples from varied locations in the Philippines in our surveillance study, indicating that leptospires might have a life cycle that does not involve animal hosts. However, despite previous work, it has not been confirmed whether leptospires multiply in the soil environment under various experimental conditions. Given the fact that the case number of leptospirosis is increased after flood, we hypothesized that waterlogged soil, which mimics the postflooding environment, could be a suitable condition for growing leptospires. To verify this hypothesis, pathogenic and saprophytic leptospires were seeded in the bottles containing 2.5 times as much water as soil, and bacterial counts in the bottles were measured over time. Pathogenic and saprophytic leptospires were found to increase their number in waterlogged soil but not in water or soil alone. In addition, leptospires were reisolated from soil in closed tubes for as long as 379 days. These results indicate that leptospires are in a resting state in the soil and are able to proliferate with increased water content in the environment. This notion is strongly supported by observations that the case number of leptospirosis is significantly higher in rainy seasons and increased after flood. Therefore, we reached the following conclusion: environmental soil is a potential reservoir of leptospires. IMPORTANCE Since research on Leptospira has focused on pathogenic leptospires, which are supposed to multiply only in animal hosts, the life cycle of saprophytic leptospires has long been a mystery. This study demonstrates that both pathogenic and saprophytic leptospires multiply in the waterlogged soil, which mimics the postflooding environment. The present results potentially explain why leptospirosis frequently occurs after floods. Therefore, environmental soil is a potential reservoir of leptospires and leptospirosis is considered an environment-borne as well as a zoonotic disease. This is a significant report to reveal that leptospires multiply under environmental conditions, and this finding leads us to reconsider the ecology of leptospires.

Characterization of the In Vitro and In Vivo Efficacy of Baloxavir Marboxil against H5 Highly Pathogenic Avian Influenza Virus Infection.
Keiichi Taniguchi, Yoshinori Ando, Masanori Kobayashi, Shinsuke Toba, Haruaki Nobori, Takao Sanaki, Takeshi Noshi, Makoto Kawai, Ryu Yoshida, Akihiko Sato, Takao Shishido, Akira Naito, Keita Matsuno, Masatoshi Okamatsu, Yoshihiro Sakoda, Hiroshi Kida
Viruses, 14, 1, 2022年01月08日, ［国際誌］
英語, 研究論文（学術雑誌）, Human infections caused by the H5 highly pathogenic avian influenza virus (HPAIV) sporadically threaten public health. The susceptibility of HPAIVs to baloxavir acid (BXA), a new class of inhibitors for the influenza virus cap-dependent endonuclease, has been confirmed in vitro, but it has not yet been fully characterized. Here, the efficacy of BXA against HPAIVs, including recent H5N8 variants, was assessed in vitro. The antiviral efficacy of baloxavir marboxil (BXM) in H5N1 virus-infected mice was also investigated. BXA exhibited similar in vitro activities against H5N1, H5N6, and H5N8 variants tested in comparison with seasonal and other zoonotic strains. Compared with oseltamivir phosphate (OSP), BXM monotherapy in mice infected with the H5N1 HPAIV clinical isolate, the A/Hong Kong/483/1997 strain, also caused a significant reduction in viral titers in the lungs, brains, and kidneys, thereby preventing acute lung inflammation and reducing mortality. Furthermore, compared with BXM or OSP monotherapy, combination treatments with BXM and OSP using a 48-h delayed treatment model showed a more potent effect on viral replication in the organs, accompanied by improved survival. In conclusion, BXM has a potent antiviral efficacy against H5 HPAIV infections.

Critical role of oxidized LDL receptor-1 in intravascular thrombosis in a severe influenza mouse model
Marumi Ohno, Akemi Kakino, Toshiki Sekiya, Naoki Nomura, Masashi Shingai, Tatsuya Sawamura, Hiroshi Kida
Scientific Reports, 11, 1, 2021年12月
研究論文（学術雑誌）, Although coagulation abnormalities, including microvascular thrombosis, are thought to contribute to tissue injury and single- or multiple-organ dysfunction in severe influenza, the detailed mechanisms have yet been clarified. This study evaluated influenza-associated abnormal blood coagulation utilizing a severe influenza mouse model. After infecting C57BL/6 male mice with intranasal applications of 500 plaque-forming units of influenza virus A/Puerto Rico/8/34 (H1N1; PR8), an elevated serum level of prothrombin fragment 1 + 2, an indicator for activated thrombin generation, was observed. Also, an increased gene expression of oxidized low-density lipoprotein (LDL) receptor-1 (Olr1), a key molecule in endothelial dysfunction in the progression of atherosclerosis, was detected in the aorta of infected mice. Body weight decrease, serum levels of cytokines and chemokines, viral load, and inflammation in the lungs of infected animals were similar between wild-type and Olr1 knockout (KO) mice. In contrast, the elevation of prothrombin fragment 1 + 2 levels in the sera and intravascular thrombosis in the lungs by PR8 virus infection were not induced in KO mice. Collectively, the results indicated that OLR1 is a critical host factor in intravascular thrombosis as a pathogeny of severe influenza. Thus, OLR1 is a promising novel therapeutic target for thrombosis during severe influenza.

Macrocyclic peptides exhibit antiviral effects against influenza virus HA and prevent pneumonia in animal models
Makoto Saito, Yasushi Itoh, Fumihiko Yasui, Tsubasa Munakata, Daisuke Yamane, Makoto Ozawa, Risa Ito, Takayuki Katoh, Hirohito Ishigaki, Misako Nakayama, Shintaro Shichinohe, Kenzaburo Yamaji, Naoki Yamamoto, Ai Ikejiri, Tomoko Honda, Takahiro Sanada, Yoshihiro Sakoda, Hiroshi Kida, Thi Quynh Mai Le, Yoshihiro Kawaoka, Kazumasa Ogasawara, Kyoko Tsukiyama-Kohara, Hiroaki Suga, Michinori Kohara
Nature Communications, 12, 1, Springer Science and Business Media LLC, 2021年12月
研究論文（学術雑誌）, Most anti-influenza drugs currently used, such as oseltamivir and zanamivir, inhibit the enzymatic activity of neuraminidase. However, neuraminidase inhibitor-resistant viruses have already been identified from various influenza virus isolates. Here, we report the development of a class of macrocyclic peptides that bind the influenza viral envelope protein hemagglutinin, named iHA. Of 28 iHAs examined, iHA-24 and iHA-100 have inhibitory effects on the in vitro replication of a wide range of Group 1 influenza viruses. In particular, iHA-100 bifunctionally inhibits hemagglutinin-mediated adsorption and membrane fusion through binding to the stalk domain of hemagglutinin. Moreover, iHA-100 shows powerful efficacy in inhibiting the growth of highly pathogenic influenza viruses and preventing severe pneumonia at later stages of infection in mouse and non-human primate cynomolgus macaque models. This study shows the potential for developing cyclic peptides that can be produced more efficiently than antibodies and have multiple functions as next-generation, mid-sized biomolecules.

Abnormal blood coagulation and kidney damage in aged hamsters infected with severe acute respiratory syndrome coronavirus 2
Marumi Ohno, Michihito Sasaki, Yasuko Orba, Toshiki Sekiya, Md Abdul Masum, Osamu Ichii, Tatsuya Sawamura, Akemi Kakino, Yasuhiko Suzuki, Hiroshi Kida, Hirofumi Sawa, Masashi Shingai
Viruses, 13, 11, 2021年11月
研究論文（学術雑誌）, Systemic symptoms have often been observed in patients with coronavirus disease 2019 (COVID-19) in addition to pneumonia, however, the details are still unclear due to the lack of an appropriate animal model. In this study, we investigated and compared blood coagulation abnormalities and tissue damage between male Syrian hamsters of 9 (young) and over 36 (aged) weeks old after intranasal infection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Despite similar levels of viral replication and inflammatory responses in the lungs of both age groups, aged but not young hamsters showed significant prolongation of prothrombin time and prominent acute kidney damage. Moreover, aged hamsters demonstrated increased intravascular coagulation time-dependently in the lungs, suggesting that consumption of coagulation factors causes prothrombin time prolongation. Furthermore, proximal urinary tract damage and mesangial matrix expansion were observed in the kidneys of the aged hamsters at early and later disease stages, respectively. Given that the severity and mortality of COVID-19 are higher in elderly human patients, the effect of aging on pathogenesis needs to be understood and should be considered for the selection of animal models. We, thus, propose that the aged hamster is a good small animal model for COVID-19 research.

Sulfated glycans containing NeuAcα2-3Gal facilitate the propagation of human H1N1 influenza A viruses in eggs
Tomomi Ichimiya, Masatoshi Okamatsu, Takaaki Kinoshita, Daiki Kobayashi, Osamu Ichii, Naoki Yamamoto, Yoshihiro Sakoda, Hiroshi Kida, Hiroto Kawashima, Kazuo Yamamoto, Sayaka Takase-Yoden, Shoko Nishihara
Virology, 562, 29, 39, Elsevier BV, 2021年10月
研究論文（学術雑誌）

Potent priming by inactivated whole influenza virus particle vaccines is linked to viral RNA uptake into antigen presenting cells.
Masashi Shingai, Naoki Nomura, Toshiki Sekiya, Marumi Ohno, Daisuke Fujikura, Chimuka Handabile, Ryosuke Omori, Yuki Ohara, Tomohiro Nishimura, Masafumi Endo, Kazuhiko Kimachi, Ryotarou Mitsumata, Tomio Ikeda, Hiroki Kitayama, Hironori Hatanaka, Tomoyoshi Sobue, Fumihito Muro, Saori Suzuki, Cong Thanh Nguyen, Hirohito Ishigaki, Misako Nakayama, Yuya Mori, Yasushi Itoh, Marios Koutsakos, Brendon Y Chua, Katherine Kedzierska, Lorena E Brown, David C Jackson, Kazumasa Ogasawara, Yoichiro Kino, Hiroshi Kida
Vaccine, 39, 29, 3940, 3951, 2021年06月29日, ［国際誌］
英語, 研究論文（学術雑誌）, Current detergent or ether-disrupted split vaccines (SVs) for influenza do not always induce adequate immune responses, especially in young children. This contrasts with the whole virus particle vaccines (WPVs) originally used against influenza that were immunogenic in both adults and children but were replaced by SV in the 1970s due to concerns with reactogenicity. In this study, we re-evaluated the immunogenicity of WPV and SV, prepared from the same batch of purified influenza virus, in cynomolgus macaques and confirmed that WPV is superior to SV in priming potency. In addition, we compared the ability of WPV and SV to induce innate immune responses, including the maturation of dendritic cells (DCs) in vitro. WPV stimulated greater production of inflammatory cytokines and type-I interferon in immune cells from mice and macaques compared to SV. Since these innate responses are likely triggered by the activation of pattern recognition receptors (PRRs) by viral RNA, the quantity and quality of viral RNA in each vaccine were assessed. Although the quantity of viral RNA was similar in the two vaccines, the amount of viral RNA of a length that can be recognized by PRRs was over 100-fold greater in WPV than in SV. More importantly, 1000-fold more viral RNA was delivered to DCs by WPV than by SV when exposed to preparations containing the same amount of HA protein. Furthermore, WPV induced up-regulation of the DC maturation marker CD86 on murine DCs, while SV did not. The present results suggest that the activation of antigen-presenting DCs, by PRR-recognizable viral RNA contained in WPV is responsible for the effective priming potency of WPV observed in naïve mice and macaques. WPV is thus recommended as an alternative option for seasonal influenza vaccines, especially for children.

The clinically used serine protease inhibitor nafamostat reduces influenza virus replication and cytokine production in human airway epithelial cells and viral replication in mice.
Mutsuo Yamaya, Yoshitaka Shimotai, Ayako Ohkawara, Enkhbold Bazarragchaa, Masatoshi Okamatsu, Yoshihiro Sakoda, Hiroshi Kida, Hidekazu Nishimura
Journal of medical virology, 93, 6, 3484, 3495, 2021年06月, ［国際誌］
英語, 研究論文（学術雑誌）, The effects of the clinically used protease inhibitor nafamostat on influenza virus replication have not been well studied. Primary human tracheal (HTE) and nasal (HNE) epithelial cells were pretreated with nafamostat and infected with the 2009 pandemic [A/Sendai-H/108/2009/(H1N1) pdm09] or seasonal [A/New York/55/2004(H3N2)] influenza virus. Pretreatment with nafamostat reduced the titers of the pandemic and seasonal influenza viruses and the secretion of inflammatory cytokines, including interleukin-6 and tumor necrosis factor-α, in the supernatants of the cells infected with the pandemic influenza virus. HTE and HNE cells exhibited mRNA and/or protein expression of transmembrane protease serine 2 (TMPRSS2), TMPRSS4, and TMPRSS11D. Pretreatment with nafamostat reduced cleavage of the precursor protein HA0 of the pandemic influenza virus into subunit HA1 in HTE cells and reduced the number of acidic endosomes in HTE and HNE cells where influenza virus RNA enters the cytoplasm. Additionally, nafamostat (30 mg/kg/day, intraperitoneal administration) reduced the levels of the pandemic influenza virus [A/Hyogo/YS/2011 (H1N1) pdm09] in mouse lung washes. These findings suggest that nafamostat may inhibit influenza virus replication in human airway epithelial cells and mouse lungs and reduce infection-induced airway inflammation by modulating cytokine production.

Updating the influenza virus library at Hokkaido University -It's potential for the use of pandemic vaccine strain candidates and diagnosis.
Naoki Nomura, Keita Matsuno, Masashi Shingai, Marumi Ohno, Toshiki Sekiya, Ryosuke Omori, Yoshihiro Sakoda, Robert G Webster, Hiroshi Kida
Virology, 557, 55, 61, 2021年05月, ［国際誌］
英語, 研究論文（学術雑誌）, Genetic reassortment of influenza A viruses through cross-species transmission contributes to the generation of pandemic influenza viruses. To provide information on the ecology of influenza viruses, we have been conducting a global surveillance of zoonotic influenza and establishing an influenza virus library. Of 4580 influenza virus strains in the library, 3891 have been isolated from over 70 different bird species. The remaining 689 strains were isolated from humans, pigs, horses, seal, whale, and the environment. Phylogenetic analyses of the HA genes of the library isolates demonstrate that the library strains are distributed to all major known clusters of the H1, H2 and H3 subtypes of HA genes that are prevalent in humans. Since past pandemic influenza viruses are most likely genetic reassortants of zoonotic and seasonal influenza viruses, a vast collection of influenza A virus strains from various hosts should be useful for vaccine preparation and diagnosis for future pandemics.

H5N1高病原性鳥インフルエンザウイルスの膜透過性ペプチドを介した細胞侵入機構　　　　　　　　　　　　　　　
梶原 直樹, 山本 直樹, 小原 道法, 安井 文彦, 迫田 義博, 喜田 宏, 芝崎 太
日本薬学会年会要旨集, 141年会, 28P01, L006, (公社)日本薬学会, 2021年03月
日本語

Efficacy of a Cap-Dependent Endonuclease Inhibitor and Neuraminidase Inhibitors against H7N9 Highly Pathogenic Avian Influenza Virus Causing Severe Viral Pneumonia in Cynomolgus Macaques.
Saori Suzuki, Cong Thanh Nguyen, Ayako Ogata-Nakahara, Akihiro Shibata, Hiroyuki Osaka, Hirohito Ishigaki, Masatoshi Okamatsu, Yoshihiro Sakoda, Hiroshi Kida, Kazumasa Ogasawara, Yasushi Itoh
Antimicrobial agents and chemotherapy, 65, 3, 2021年02月17日, ［国際誌］
英語, 研究論文（学術雑誌）, H7N9 highly pathogenic avian influenza virus (HPAIV) infection in a human was first reported in 2017. A/duck/Japan/AQ-HE29-22/2017 (H7N9) (Dk/HE29-22), found in imported duck meat at an airport in Japan, possesses a hemagglutinin with a multibasic cleavage site, indicating high pathogenicity in chickens, as in the case of other H7 HPAIVs. In the present study, we examined the pathogenicity of Dk/HE29-22 and the effectiveness of a cap-dependent endonuclease inhibitor (baloxavir) and neuraminidase inhibitors (oseltamivir and zanamivir) against infection with this strain in a macaque model (n = 3 for each group). All of the macaques infected with Dk/HE29-22 showed severe signs of disease and pneumonia even after the virus had disappeared from lung samples. Virus titers in macaques treated with baloxavir were significantly lower than those in the other treated groups. After infection, levels of interferon alpha and beta (IFN-α and IFN-β) in the blood of macaques in the baloxavir group were the highest among the groups, whereas levels of tumor necrosis factor alpha (TNF-α) and interleukin 13 (IL-13) were slightly increased in the untreated group. In addition, immune checkpoint proteins, including programmed death 1 (PD-1) and T cell immunoreceptor with Ig and ITIM domains (TIGIT), were expressed at high levels in the untreated group, especially in one macaque that showed severe signs of disease, indicating that negative feedback responses against vigorous inflammation may contribute to disease progression. In the group treated with baloxavir, the percentages of PD-1-, CTLA-4-, and TIGIT-positive T lymphocytes were lower than those in the untreated group, indicating that reduction in virus titers may prevent expression of immune checkpoint molecules from downregulation of T cell responses.

Potency of an Inactivated Influenza Vaccine against a Challenge with A/Swine/Missouri/A01727926/2015 (H4N6) in Mice for Pandemic Preparedness.
Hirotaka Hayashi, Norikazu Isoda, Enkhbold Bazarragchaa, Naoki Nomura, Keita Matsuno, Masatoshi Okamatsu, Hiroshi Kida, Yoshihiro Sakoda
Vaccines, 8, 4, 2020年12月16日, ［国際誌］
英語, 研究論文（学術雑誌）, H4 influenza viruses have been isolated from birds across the world. In recent years, an H4 influenza virus infection has been confirmed in pigs. Pigs play an important role in the transmission of influenza viruses to human hosts. Therefore, it is important to develop a new vaccine in the case of an H4 influenza virus infection in humans, considering that this virus has a different antigenicity from seasonal human influenza viruses. In this study, after selecting vaccine candidate strains based on their antigenic relation to one of the pig isolates, A/swine/Missouri/A01727926/2015 (H4N6) (MO/15), an inactivated whole-particle vaccine was prepared from A/swan/Hokkaido/481102/2017 (H4N6). This vaccine showed high immunogenicity in mice, and the antibody induced by the vaccine showed high cross-reactivity to the MO/15 virus. This vaccine induced sufficient neutralizing antibodies and mitigated the effects of an MO/15 infection in a mouse model. This study is the first to suggest that an inactivated whole-particle vaccine prepared from an influenza virus isolated from wild birds is an effective countermeasure in case of a future influenza pandemic caused by the H4 influenza virus.

Re-Invasion of H5N8 High Pathogenicity Avian Influenza Virus Clade 2.3.4.4b in Hokkaido, Japan, 2020.
Norikazu Isoda, Augustin T Twabela, Enkhbold Bazarragchaa, Kohei Ogasawara, Hirotaka Hayashi, Zu-Jyun Wang, Daiki Kobayashi, Yukiko Watanabe, Keisuke Saito, Hiroshi Kida, Yoshihiro Sakoda
Viruses, 12, 12, 2020年12月14日, ［国際誌］
英語, 研究論文（学術雑誌）, Global dispersion of high pathogenicity avian influenza (HPAI), especially that caused by H5 clade 2.3.4.4, has threatened poultry industries and, potentially, human health. An HPAI virus, A/northern pintail/Hokkaido/M13/2020 (H5N8) (NP/Hok/20) belonging to clade 2.3.4.4b, was isolated from a fecal sample collected at a lake in Hokkaido, Japan where migratory birds rested, October 2020. In the phylogenetic trees of all eight gene segments, NP/Hok/20 fell into in the cluster of European isolates in 2020, but was distinct from the isolates in eastern Asia and Europe during the winter season of 2017-2018. The antigenic cartography indicates that the antigenicity of NP/Hok/20 was almost the same as that of previous isolates of H5 clade 2.3.4.4b, whereas the antigenic distances from NP/Hok/20 to the representative strains in clade 2.3.4.4e and to a strain in 2.3.4 were apparently distant. These data imply that HPAI virus clade 2.3.4.4b should have been delivered by bird migration despite the intercontinental distance, although it was not defined whether NP/Hok/20 was transported from Europe via Siberia where migratory birds nest in the summer season. Given the probability of perpetuation of transmission in the northern territory, periodic updates of intensive surveys on avian influenza at the global level are essential to prepare for future outbreaks of the HPAI virus.

In vitro anti-inflammatory and anti-lipid accumulation properties of taxifolin-rich extract from the Japanese larch, Larix kaempferi
Daisuke Muramatsu, Hirofumi Uchiyama, Hiroshi Kida, Atsushi Iwai
Heliyon, 6, 12, e05505, e05505, Elsevier BV, 2020年12月
研究論文（学術雑誌）

Cell-penetrating peptide-mediated cell entry of H5N1 highly pathogenic avian influenza virus
Naoki Kajiwara, Namiko Nomura, Masako Ukaji, Naoki Yamamoto, Michinori Kohara, Fumihiko Yasui, Yoshihiro Sakoda, Hiroshi Kida, Futoshi Shibasaki
Scientific Reports, 10, 1, Springer Science and Business Media LLC, 2020年12月
研究論文（学術雑誌）,
H5N1 highly pathogenic avian influenza virus (HPAIV) poses a huge threat to public health and the global economy. These viruses cause systemic infection in poultry and accidental human infection leads to severe pneumonia, associated with high mortality rates. The hemagglutinin (HA) of H5N1 HPAIV possesses multiple basic amino acids, as in the sequence RERRRKKR at the cleavage site; however, the role of this motif is not fully understood. Here, we showed that a 33-amino acid long peptide derived from HA of H5N1 HPAIV (HA314-46) has the potential to penetrate various cells and lung tissue through a sialic acid-independent endocytotic pathway. Mutant peptide analyses revealed that the cysteine residue at position 318 and multiple basic amino acids were essential for the cell-penetrating activity. Moreover, reassortant viruses possessing H5 HA could enter sialic acid-deficient cells, and virus internalisation was facilitated by cleavage with recombinant furin. Thus, our findings demonstrate that the HA314-46 motif exhibits cell-penetrating activity through a sialic acid-independent cell entry mechanism.

Influenza virus infection affects insulin signaling, fatty acid-metabolizing enzyme expressions, and the tricarboxylic acid cycle in mice
Marumi Ohno, Toshiki Sekiya, Naoki Nomura, Taku ji Daito, Masashi Shingai, Hiroshi Kida
Scientific Reports, 10, 1, Springer Science and Business Media LLC, 2020年12月
研究論文（学術雑誌）

2016-2017年のベトナム南部での疫学的研究から明らかになった、鳥インフルエンザウイルスの新たなホットスポットの意義(The implication of a new hot spot of avian influenza based on epidemiological study in southern Vietnam in 2016-2017)　　　　　　　　　　　　　　　
Le Trung Kien, 磯田 典和, Nguyen Thanh Lam, Chu Duc-Huy, Tien Tien Ngoc, 松野 啓太, 岡松 正敏, 喜田 宏, 迫田 義博, 北海道大人獣共通感染症リサーチセンター/国際協働ユニット
日本獣医学会学術集会講演要旨集, 163回, 232, 232, (公社)日本獣医学会, 2020年10月
英語

Immune profiling of influenza‐specific B‐ and T‐cell responses in macaques using flow cytometry‐based assays
Marios Koutsakos, Toshiki Sekiya, Brendon Y Chua, Thi Hoang Oanh Nguyen, Adam K Wheatley, Jennifer A Juno, Marumi Ohno, Naoki Nomura, Yuki Ohara, Tomohiro Nishimura, Masafumi Endo, Saori Suzuki, Hirohito Ishigaki, Misako Nakayama, Cong T Nguyen, Yasushi Itoh, Masashi Shingai, Kazumasa Ogasawara, Yoichiro Kino, Stephen J Kent, David C Jackson, Lorena E Brown, Hiroshi Kida, Katherine Kedzierska
Immunology & Cell Biology, Wiley, 2020年09月07日
研究論文（学術雑誌）

Genetic and antigenic characterization of H5 and H7 avian influenza viruses isolated from migratory waterfowl in Mongolia from 2017 to 2019.
Ankhanbaatar Ulaankhuu, Enkhbold Bazarragchaa, Masatoshi Okamatsu, Takahiro Hiono, Khishgee Bodisaikhan, Tsolmon Amartuvshin, Jargalsaikhan Tserenjav, Tsogtbaatar Urangoo, Khanui Buyantogtokh, Keita Matsuno, Takanari Hattori, Tatsunari Kondoh, Masahiro Sato, Yoshihiro Takadate, Shiho Torii, Mao Isono, Kosuke Okuya, Takeshi Saito, Nodoka Kasajima, Yurie Kida, Junki Maruyama, Manabu Igarashi, Ayato Takada, Hiroshi Kida, Damdinjav Batchuluun, Yoshihiro Sakoda
Virus genes, 56, 4, 472, 479, 2020年08月, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, The circulation of highly pathogenic avian influenza viruses (HPAIVs) of various subtypes (e.g., H5N1, H5N6, H5N8, and H7N9) in poultry remains a global concern for animal and public health. Migratory waterfowls play important roles in the transmission of these viruses across countries. To monitor virus spread by wild birds, active surveillance for avian influenza in migratory waterfowl was conducted in Mongolia from 2015 to 2019. In total, 5000 fecal samples were collected from lakesides in central Mongolia, and 167 influenza A viruses were isolated. Two H5N3, four H7N3, and two H7N7 viruses were characterized in this study. The amino acid sequence at hemagglutinin (HA) cleavage site of those isolates suggested low pathogenicity in chickens. Phylogenetic analysis revealed that all H5 and H7 viruses were closely related to recent H5 and H7 low pathogenic avian influenza viruses (LPAIVs) isolated from wild birds in Asia and Europe. Antigenicity of H7Nx was similar to those of typical non-pathogenic avian influenza viruses (AIVs). While HPAIVs or A/Anhui/1/2013 (H7N9)-related LPAIVs were not detected in migratory waterfowl in Mongolia, sporadic introductions of AIVs including H5 and H7 viruses into Mongolia through the wild bird migration were identified. Thus, continued monitoring of H5 and H7 AIVs in both domestic and wild birds is needed for the early detection of HPAIVs spread into the country.

Efficacy of neuraminidase inhibitors against H5N6 highly pathogenic avian influenza virus in a non-human primate model.
Cong Thanh Nguyen, Saori Suzuki, Yasushi Itoh, Hirohito Ishigaki, Misako Nakayama, Kaori Hayashi, Keita Matsuno, Masatoshi Okamatsu, Yoshihiro Sakoda, Hiroshi Kida, Kazumasa Ogasawara
Antimicrobial agents and chemotherapy, 2020年04月13日, ［査読有り］, ［国際誌］
英語, Attention has been paid to H5N6 highly pathogenic avian influenza virus (HPAIV) because of its heavy burden on the poultry industry and human mortality. Since an influenza A virus carrying N6 neuraminidase (NA) has never spread in humans, the potential for H5N6 HPAIV to cause disease in humans and the efficacy of antiviral drugs against the virus need to be urgently assessed. We used non-human primates to elucidate the pathogenesis of H5N6 HPAIV as well as to determine the efficacy of antiviral drugs against the virus. H5N6 HPAIV infection led to high fever in cynomolgus macaques. The lung injury caused by the virus was severe with diffuse alveolar damage and neutrophil infiltration. In addition, an increase in IFN-α showed an inverse correlation with virus titers during the infection process. Oseltamivir was effective for reducing H5N6 HPAIV propagation, and continuous treatment with peramivir reduced virus propagation and severity of symptoms in the early stage. This study also showed the pathologically severe lung injury states in the cynomolgus macaques infected with H5N6 HPAIV, even in those that received early antiviral drug treatments, indicating the need for close monitoring and the need for further studies on the virus pathogenicity and new antiviral therapies.

Low replicative fitness of neuraminidase inhibitor-resistant H7N9 avian influenza a virus with R292K substitution in neuraminidase in cynomolgus macaques compared with I222T substitution.
Saori Suzuki, Shintaro Shichinohe, Yasushi Itoh, Misako Nakayama, Hirohito Ishigaki, Yuya Mori, Ayako Ogata-Nakahara, Cong Thanh Nguyen, Masatoshi Okamatsu, Yoshihiro Sakoda, Hiroshi Kida, Kazumasa Ogasawara
Antiviral research, 178, 104790, 104790, 2020年04月06日, ［査読有り］, ［国際誌］
英語, Human cases of H7N9 influenza A virus infection have been increasing since 2013. The first choice of treatment for influenza is neuraminidase (NA) inhibitors (NAIs), but there is a concern that NAI-resistant viruses are selected in the presence of NAIs. In our previous study, an H7N9 virus carrying AA substitution of threonine (T) for isoleucine (I) at residue 222 in NA (NA222T, N2 numbering) and an H7N9 virus carrying AA substitution of lysine (K) for arginine (R) at residue 292 in NA (NA292K, N2 numbering) were found in different macaques that had been infected with A/Anhui/1/2013 (H7N9) and treated with NAIs. In the present study, the variant with NA292K showed not only resistance to NAIs but also lower replication activity in MDCK cells than did the virus with wild-type NA, whereas the variant with NA222T, which was less resistant to NAIs, showed replication activity similar to that of the wild-type virus. Next, we examined the pathogenicity of these H7N9 NAI-resistant viruses in macaques. The variants caused clinical signs similar to those caused by the wild-type virus with similar replication potency. However, the virus with NA292K was replaced within 7 days by that with NA292R (same as the wild-type) in nasal samples from macaques infected with the virus with NA292K, i.e. the so-called revertant (wild-type virus) became dominant in the population in the absence of an NAI. These results suggest that the clinical signs observed in macaques infected with the NA292K virus are caused by the NA292K virus and the NA292R virus and that the virus with NA292K may not replicate continuously in the upper respiratory tract of patients without treatment as effectively as the wild-type virus.

Genetic and antigenic characterization of the first H7N7 low pathogenic avian influenza viruses isolated in Vietnam.
Kien Trung Le, Masatoshi Okamatsu, Lam Thanh Nguyen, Keita Matsuno, Duc-Huy Chu, Tien Ngoc Tien, Tung Thanh Le, Hiroshi Kida, Yoshihiro Sakoda
Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases, 78, 104117, 104117, 2020年03月, ［査読有り］, ［国際誌］
英語, During the annual surveillance of avian influenza viruses (AIVs) in Vietnam in 2018, three H7N7 AIV isolates were identified in domestic ducks in a single flock in Vinh Long province. The present study is the first documented report of H7N7 virus isolates in Vietnam and aimed to characterize these viruses, both genetically and antigenically. Deduced amino acid sequences for the hemagglutinins (HAs) indicated a low pathogenicity of these viruses in chickens. Phylogenetic analysis revealed that the H7 HA genes of these isolates were closely related to each other and belonged to the European-Asian sublineage, together with those of H7N3 viruses isolated from ducks in Cambodia during 2017. They were not genetically related to those of Chinese H7N9 or H7N1 viruses that were previously detected in Vietnam during 2012. Interestingly, the M genes of the two H7N7 virus isolates were phylogenetically classified into distinct groups, suggesting an ongoing reassortment event in domestic ducks because they were isolated from the same flock. These H7N7 viruses exhibited somewhat different antigenic characteristics compared with other representative H7 low pathogenic AIVs. Surprisingly, the antigenicity of Vietnamese H7N7 viruses is similar to Chinese H7N9 highly pathogenic AIV. The findings of this study suggest that H7N7 viruses may be undergoing reassortment and antigenic diversification in poultry flocks in Vietnam. The silent spread of Vietnamese H7N7 viruses in chickens may lead to acquire high pathogenicity in chickens although the zoonotic potential of the viruses seems to be low since these viruses retain typical avian-specific motifs in the receptor-binding site in the HA and there is no mutation related to mammalian adaptation in PB2 gene. Thus, these results highlight the need for continuous and intensive surveillance of avian influenza in Vietnam, targeting not only highly pathogenic AIVs but also low pathogenic viruses.

A cloned classical swine fever virus derived from the vaccine strain GPE- causes cytopathic effect in CPK-NS cells via type-I interferon-dependent necroptosis.
Yukari Itakura, Keita Matsuno, Asako Ito, Markus Gerber, Matthias Liniger, Yuri Fujimoto, Tomokazu Tamura, Ken-Ichiro Kameyama, Masatoshi Okamatsu, Nicolas Ruggli, Hiroshi Kida, Yoshihiro Sakoda
Virus research, 276, 197809, 197809, 2020年01月15日, ［査読有り］, ［国際誌］
英語, Classical swine fever viruses (CSFVs) do typically not show cytopathic effect (CPE) in cell culture, while some strains such as vaccine strain the GPE- induce CPE in the swine kidney-derived CPK-NS cell line cultured in serum-free medium. These latter strains commonly lack Npro-mediated inhibition of type-I interferon (IFN) induction. In order to explore the molecular mechanisms of GPE--induced CPE, we analyzed the cellular pathways involved. In CPK-NS cells infected with the attenuated-vaccine-derived vGPE- strain, both, apoptosis and necroptosis were induced. Necroptosis was type-I IFN-dependent and critical for visible CPE. In contrast, the parental virulent vALD-A76 strain did not induce any of these pathways nor CPE. We used reverse genetics to investigate which viral factors regulate these cell-death pathways. Interestingly, a mutant vGPE- in which the Npro function was restored to inhibit type-I IFN induction did not induce necroptosis nor CPE but still induced apoptosis, while an Npro-mutant vALD-A76 incapable of inhibiting type-I IFN production induced necroptosis and CPE. Although Erns of CSFV is reportedly involved in controlling apoptosis, apoptosis induction by vGPE- or apoptosis inhibition by vALD-A76 were independent of the unique amino acid difference found in Erns of these two strains. Altogether, these results demonstrate that type-I IFN-dependent necroptosis related to non-functional Npro is the main mechanism for CPE induction by vGPE-, and that viral factor(s) other than Erns may induce or inhibit apoptosis in vGPE- or vALD-A76 infected CPK-NS cells, respectively.

E190V substitution of H6 hemagglutinin is one of key factors for binding to sulfated sialylated glycan receptor and infection to chickens.
Yuto Kikutani, Masatoshi Okamatsu, Shoko Nishihara, Sayaka Takase-Yoden, Takahiro Hiono, Robert P de Vries, Ryan McBride, Keita Matsuno, Hiroshi Kida, Yoshihiro Sakoda
Microbiology and immunology, 2020年01月14日, ［査読有り］, ［国際誌］
英語, Avian influenza viruses (AIVs) recognize sialic acid linked α2,3 to galactose (SAα2,3Gal) glycans as receptors. In this study, the interactions between hemagglutinins (HAs) of AIVs and sulfated SAα2,3Gal glycans were analyzed to clarify the molecular basis of interspecies transmission of AIVs from ducks to chickens. It was revealed that E190V and N192D substitutions of the HA increased the recovery of viruses derived from an H6 duck virus isolate, A/duck/Hong Kong/960/1980 (H6N2), in chickens. Recombinant HAs from an H6 chicken virus, A/chicken/Tainan/V156/1999 (H6N1), bound to sulfated SAα2,3Gal glycans, whereas the HAs from an H6 duck virus did not. Binding preference of mutant HAs revealed that an E190V substitution is critical for the recognition of sulfated SAα2,3Gal glycans. These results suggest that the binding of the HA from H6 AIVs to sulfated SAα2,3Gal glycans explains a part of mechanisms of interspecies transmission of AIVs from ducks to chickens.

ベトナムで初単離されたH7N7低病原性鳥インフルエンザウイルスの遺伝学的性状および抗原性の解析(Genetic and antigenic characterization of H7N7 low pathogenic avian influenza viruses firstly isolated in Vietnam)　　　　　　　　　　　　　　　
Le Trung Kien, Nguyen Thanh Lam, Chu Duc-Huy, Nguyen Tien Ngoc, Nguyen Long Van, Tien Tien Ngoc, 松野 啓太, 岡松 正敏, 喜田 宏, 迫田 義博
日本獣医学会学術集会講演要旨集, 162回, 392, 392, (公社)日本獣医学会, 2019年08月
英語

Cell cytotoxity and anti-glycation activity of taxifolin-rich extract from Japanese larch, Larix kaempferi.
Muramatsu D, Uchiyama H, Kida H, Iwai A
Heliyon, 5, 7, e02047, 2019年07月, ［査読有り］

A systematic study towards evolutionary and epidemiological dynamics of currently predominant H5 highly pathogenic avian influenza viruses in Vietnam.
Lam Thanh Nguyen, Simon M Firestone, Mark A Stevenson, Neil D Young, Leslie D Sims, Duc Huy Chu, Tien Ngoc Nguyen, Long Van Nguyen, Tung Thanh Le, Hung Van Nguyen, Hung Nam Nguyen, Tien Ngoc Tien, Tho Dang Nguyen, Bich Ngoc Tran, Keita Matsuno, Masatoshi Okamatsu, Hiroshi Kida, Yoshihiro Sakoda
Scientific reports, 9, 1, 7723, 7723, 2019年05月22日, ［査読有り］, ［国際誌］
英語, This study aimed to elucidate virus, host and environmental dynamics of Vietnamese H5 highly pathogenic avian influenza viruses (HPAIVs) during 2014-2017. Epidemiologically, H5 HPAIVs were frequently detected in apparently healthy domestic and Muscovy ducks and therefore these are preferred species for H5 HPAIV detection in active surveillance. Virologically, clade 2.3.2.1c and 2.3.4.4 H5 HPAIVs were predominant and exhibited distinct phylogeographic evolution. Clade 2.3.2.1c viruses clustered phylogenetically in North, Central and South regions, whilst clade 2.3.4.4 viruses only detected in North and Central regions formed small groups. These viruses underwent diverse reassortment with existence of at least 12 genotypes and retained typical avian-specific motifs. These H5 HPAIVs exhibited large antigenic distance from progenitor viruses and commercial vaccines currently used in poultry. Bayesian phylodynamic analysis inferred that clade 2.3.2.1c viruses detected during 2014-2017 were likely descended from homologous clade viruses imported to Vietnam previously and/or preexisting Chinese viruses during 2012-2013. Vietnamese clade 2.3.4.4 viruses closely shared genetic traits with contemporary foreign spillovers, suggesting that there existed multiple transboundary virus dispersals to Vietnam. This study provides insights into the evolution of Vietnamese H5 HPAIVs and highlights the necessity of strengthening control measures such as, preventive surveillance and poultry vaccination.

Spatiotemporal and risk analysis of H5 highly pathogenic avian influenza in Vietnam, 2014-2017.
Lam Thanh Nguyen, Mark A Stevenson, Simon M Firestone, Leslie D Sims, Duc Huy Chu, Long Van Nguyen, Tien Ngoc Nguyen, Kien Trung Le, Norikazu Isoda, Keita Matsuno, Masatoshi Okamatsu, Hiroshi Kida, Yoshihiro Sakoda
Preventive veterinary medicine, 104678, 104678, 2019年04月22日, ［査読有り］, ［国際誌］
英語, The aim of this study was to describe the spatiotemporal distribution of H5 HPAI outbreak reports for the period 2014-2017 and to identify factors associated with H5 HPAI outbreak reports. Throughout the study period, a total of 139 outbreaks of H5 HPAI in poultry were reported, due to either H5N1 (96 outbreaks) or H5N6 (43 outbreaks) subtype viruses. H5N1 HPAI outbreaks occurred in all areas of Vietnam while H5N6 HPAI outbreaks were only reported in the northern and central provinces. We counted the number of H5N1 and H5N6 outbreak report-positive districts per province over the four-year study period and calculated the provincial-level standardized morbidity ratio for H5N1 and H5N6 outbreak reports as the observed number of positive districts divided by the expected number. A mixed-effects, zero-inflated Poisson regression model was developed to identify risk factors for outbreak reports of each H5N1 and H5N6 subtype virus. Spatially correlated and uncorrelated random effects terms were included in this model to identify areas of the country where outbreak reports occurred after known risk factors had been accounted-for. The presence of an outbreak report in a province in the previous 6-12 months increased the provincial level H5N1 outbreak report risk by a factor of 2.42 (95% Bayesian credible interval [CrI] 1.27-4.60) while 1000 bird increases in the density of chickens decreased provincial level H5N6 outbreak report risk by a factor of 0.65 (95% CrI 0.38 to 0.97). We document distinctly different patterns in the spatial and temporal distribution of H5N1 and H5N6 outbreak reports. Most of the variation in H5N1 report risk was accounted-for by the fixed effects included in the zero-inflated Poisson model. In contrast, the amount of unaccounted-for risk in the H5N6 model was substantially greater than the H5N1 model. For H5N6 we recommend that targeted investigations should be carried out in provinces with relatively large spatially correlated random effect terms to identify likely determinants of disease. Similarly, investigations should be carried out in provinces with relatively low spatially correlated random effect terms to identify protective factors for disease and/or reasons for failure to report.

Inhibition of avian-origin influenza A(H7N9) virus by the novel cap-dependent endonuclease inhibitor baloxavir marboxil.
Keiichi Taniguchi, Yoshinori Ando, Haruaki Nobori, Shinsuke Toba, Takeshi Noshi, Masanori Kobayashi, Makoto Kawai, Ryu Yoshida, Akihiko Sato, Takao Shishido, Akira Naito, Keita Matsuno, Masatoshi Okamatsu, Yoshihiro Sakoda, Hiroshi Kida
Scientific reports, 9, 1, 3466, 3466, 2019年03月05日, ［査読有り］, ［国際誌］
英語, Human infections with avian-origin influenza A(H7N9) virus represent a serious threat to global health; however, treatment options are limited. Here, we show the inhibitory effects of baloxavir acid (BXA) and its prodrug baloxavir marboxil (BXM), a first-in-class cap-dependent endonuclease inhibitor, against A(H7N9), in vitro and in vivo. In cell culture, BXA at four nanomolar concentration achieved a 1.5-2.8 log reduction in virus titers of A(H7N9), including the NA-R292K mutant virus and highly pathogenic avian influenza viruses, whereas NA inhibitors or favipiravir required approximately 20-fold or higher concentrations to achieve the same levels of reduction. A(H7N9)-specific amino acid polymorphism at position 37, implicated in BXA binding to the PA endonuclease domain, did not impact on BXA susceptibility. In mice, oral administration of BXM at 5 and 50 mg/kg twice a day for 5 days completely protected from a lethal A/Anhui/1/2013 (H7N9) challenge, and reduced virus titers more than 2-3 log in the lungs. Furthermore, the potent therapeutic effects of BXM in mice were still observed when a higher virus dose was administered or treatment was delayed up to 48 hours post infection. These findings support further investigation of BXM for A(H7N9) treatment in humans.

Inactivated whole virus particle vaccine with potent immunogenicity and limited IL-6 production is ideal for influenza　　　　　　　　　　　　　　　
Toshiki Sekiya, Edin J Mifsud, Marumi Ohno, Naoki Nomura, Mayumi Sasada, Daisuke Fujikura, Takuji Daito, Masashi Shingai, Yuki Ohara, Tomohiro Nishimura, Masafumi Endo, Ryotarou Mitsumata, Tomio Ikeda, Hironori Hatanaka, Hiroki Kitayama, Kenji Motokawa, Tomoyoshi Sobue, Saori Suzuki, Yasushi Itoh, Lorena E Brown, Kazumasa Ogasawara, Yoichiro Kino, Hiroshi Kida
Vaccine, 37, 2158, 2166, 2019年03月, ［査読有り］
英語, 研究論文（学術雑誌）

Evaluation of a rapid isothermal nucleic acid amplification kit, Alere™ i Influenza A&B, for the detection of avian influenza viruses.
Bazarragchaa E, Okamatsu M, Ulaankhuu A, Twabela AT, Matsuno K, Kida H, Sakoda Y
J Virol Methods, 265, 121, 125, 2019年03月, ［査読有り］, ［国際誌］
英語, Rapid and accurate diagnosis of influenza virus infection is essential for quick responses for both human and animal health. The Alere™ i Influenza A&B is a novel isothermal nucleic acid amplification kit that can detect and differentiate between influenza A and B viruses in human specimens in approximately 15 min. In the present study, the performance of the Alere™ i Influenza A&B kit was evaluated for its ability to detect avian influenza virus in chickens. The kit was able to detect representative avian influenza virus strains (hemagglutinin subtypes H1-H16, including the recently isolated H5 and H7 highly pathogenic avian influenza viruses), and the detection limit of the kit for these viruses varied between 10-1.4-102.1 50% egg-infective dose per test, which is higher than the analytical sensitivity of the antigen detection immunochromatography kit ESPLINE® A INFLUENZA. In experimentally infected chickens inoculated with a highly pathogenic avian influenza virus strain A/chicken/Hokkaido/002/2016 (H5N6), viral RNA was detected in the tracheal and cloacal swabs. These results indicate that this kit has the potential to be used as a rapid screening test of influenza A virus infection in chickens.

In vitro characterization of baloxavir acid, a first-in-class cap-dependent endonuclease inhibitor of the influenza virus polymerase PA subunit.
Takeshi Noshi, Mitsutaka Kitano, Keiichi Taniguchi, Atsuko Yamamoto, Shinya Omoto, Keiko Baba, Takashi Hashimoto, Kayo Ishida, Yukihiro Kushima, Kazunari Hattori, Makoto Kawai, Ryu Yoshida, Masanori Kobayashi, Tomokazu Yoshinaga, Akihiko Sato, Masatoshi Okamatsu, Yoshihiro Sakoda, Hiroshi Kida, Takao Shishido, Akira Naito
Antiviral research, 160, 109, 117, 2018年12月, ［査読有り］, ［国際誌］
英語, Cap-dependent endonuclease (CEN) resides in the PA subunit of the influenza virus and mediates the critical "cap-snatching" step of viral RNA transcription, which is considered to be a promising anti-influenza target. Here, we describe in vitro characterization of a novel CEN inhibitor, baloxavir acid (BXA), the active form of baloxavir marboxil (BXM). BXA inhibits viral RNA transcription via selective inhibition of CEN activity in enzymatic assays, and inhibits viral replication in infected cells without cytotoxicity in cytopathic effect assays. The antiviral activity of BXA is also confirmed in yield reduction assays with seasonal type A and B viruses, including neuraminidase inhibitor-resistant strains. Furthermore, BXA shows broad potency against various subtypes of influenza A viruses (H1N2, H5N1, H5N2, H5N6, H7N9 and H9N2). Additionally, serial passages of the viruses in the presence of BXA result in isolation of PA/I38T variants with reduced BXA susceptibility. Phenotypic and genotypic analyses with reverse genetics demonstrate the mechanism of BXA action via CEN inhibition in infected cells. These results reveal the in vitro characteristics of BXA and support clinical use of BXM to treat influenza.

MicroRNA-451a in extracellular, blood-resident vesicles attenuates macrophage and dendritic cell responses to influenza whole-virus vaccine.
Okamoto M, Fukushima Y, Kouwaki T, Daito T, Kohara M, Kida H, Oshiumi H
The Journal of biological chemistry, 293, 48, 18585, 18600, 2018年11月, ［査読有り］
研究論文（学術雑誌）, The innate immune system is important for the efficacy of vaccines, but excessive innate immune responses can cause adverse reactions after vaccination. Extracellular vesicles (EVs) are enriched in the blood and can deliver functional RNAs, such as microRNAs (miRNAs), to recipient cells, thereby mediating intercellular communication. However, the role of EVs in controlling the innate immune responses to vaccines has not been fully elucidated. Here, we found that miR-451a is abundant in human serum EVs and that its presence in blood-circulating EVs affects the innate immune responses of macrophages and dendritic cells to inactivated whole-virus vaccines (WV) against influenza. miR-451a in human serum EVs was stable for a week in healthy subjects, and its levels gradually fluctuated over several months. miR-451a within serum EVs was internalized into serum-cultured macrophages and dendritic cells and reduced endogenous 14-3-3 protein levels and decreased the expression of type I IFN and interleukin 6 in response to WV stimulation. miR-451a levels in blood-circulating EVs were positively correlated with intracellular miR-451a levels in mouse splenic CD11c cells and inversely correlated with the innate immune response to inactivated WV in vivo. These findings suggest that miR-451a in circulating EVs is internalized into recipient cells in vivo and that this internalization results in an attenuation of the innate immune response to WV. Moreover, a microarray analysis identified several other miRNAs that affect the macrophage response to inactivated WV. Our results reveal that miRNAs in circulating EVs significantly modify the responses of macrophages and dendritic cells to inactivated WV.

Repeated detection of H7N9 avian influenza viruses in raw poultry meat illegally brought to Japan by international flight passengers.
Akihiro Shibata, Masatoshi Okamatsu, Riho Sumiyoshi, Keita Matsuno, Zu-Jyun Wang, Hiroshi Kida, Hiroyuki Osaka, Yoshihiro Sakoda
Virology, 524, 10, 17, 2018年11月, ［査読有り］, ［国際誌］
英語, H7N9 highly and low pathogenic avian influenza viruses (HPAIV and LPAIV, respectively) have been isolated from duck meat products that were brought illegally into Japan by flight passengers in their hand luggage. These H7N9 virus isolates were phylogenetically closely related to those prevailing in China. Antigenic analysis revealed that the hemagglutinin of the H7N9 HPAIV isolate was slightly different from those of the H7N9 LPAIV and older H7 strains. These meat products contaminated with AIVs repeatedly brought into Japan lead to increased risks of poultry and public health. Continuous border disease control based on the detection and culling of infected poultry and meat products is, thus, essential for the prevention of introduction and spread of AIVs.

Cervical dystonia in Parkinson's disease: Retrospective study of later-stage clinical features
Kida Hiroshi, Miura Shiroh, Yamanishi Yoshihiro, Takahashi Tomoyuki, Kamada Takashi, Yorita Akiko, Ayabe Mitsuyoshi, Kida Hideki, Hoshino Tomoaki, Taniwaki Takayuki
NEUROLOGY ASIA, 23, 3, 245, 251, 2018年09月, ［査読有り］

H13 influenza viruses in wild birds have undergone genetic and antigenic diversification in nature.
Zu-Jyun Wang, Yuto Kikutani, Lam Thanh Nguyen, Takahiro Hiono, Keita Matsuno, Masatoshi Okamatsu, Scott Krauss, Richard Webby, Youn-Jeong Lee, Hiroshi Kida, Yoshihiro Sakoda
Virus genes, 54, 4, 543, 549, 2018年08月, ［査読有り］, ［国際誌］
英語, Among 16 haemagglutinin (HA) subtypes of avian influenza viruses (AIVs), H13 AIVs have rarely been isolated in wild waterfowl. H13 AIVs cause asymptomatic infection and are maintained mainly in gull and tern populations; however, the recorded antigenic information relating to the viruses has been limited. In this study, 2 H13 AIVs, A/duck/Hokkaido/W345/2012 (H13N2) and A/duck/Hokkaido/WZ68/2012 (H13N2), isolated from the same area in the same year in our surveillance, were genetically and antigenically analyzed with 10 representative H13 strains including a prototype strain, A/gull/Maryland/704/1977 (H13N6). The HA genes of H13 AIVs were phylogenetically divided into 3 groups (I, II, and III). A/duck/Hokkaido/W345/2012 (H13N2) was genetically classified into Group III. This virus was distinct from a prototype strain, A/gull/Maryland/704/1977 (H13N6), and the virus, A/duck/Hokkaido/WZ68/2012 (H13N2), both belonging to Group I. Antigenic analysis indicated that the viruses of Group I were antigenically closely related to those of Group II, but distinct from those of Group III, including A/duck/Hokkaido/W345/2012 (H13N2). In summary, our study indicates that H13 AIVs have undergone antigenic diversification in nature.

Potency of an inactivated influenza vaccine prepared from A/duck/Mongolia/245/2015 (H10N3) against H10 influenza virus infection in a mouse model　　　　　　　　　　　　　　　
Suzuki, M, Okamatsu, M, Fujimoto, Y, Hiono, T, Matsuno, K, Kida, H, Sakoda, Y
Japanese journal of Veterinary Research, 66, 1, 29, 41, 2018年02月, ［査読有り］
英語, 研究論文（学術雑誌）

Aureobasidium pullulans-cultured fluid induces IL-18 production, leading to Th1-polarization during influenza A virus infection
Daisuke Fujikura, Daisuke Muramatsu, Kochi Toyomane, Satoko Chiba, Takuji Daito, Atsushi Iwai, Takahisa Kouwaki, Masaaki Okamoto, Hideaki Higashi, Hiroshi Kida, Hiroyuki Oshiumi
Journal of Biochemistry, 163, 1, 31, 38, Oxford University Press, 2018年01月01日, ［査読有り］
英語, 研究論文（学術雑誌）, Several microbial molecules with pathogen-associated molecular patterns stimulate host innate immune responses. The innate immune system plays a crucial role in activating acquired immune response via cytokine production and antigen presentation. Previous studies have shown that Aureobasidium pullulans-cultured fluid (AP-CF), which contains b-glucan, exhibits adjuvant activity and renders mice resistance to influenza A virus infection
however, the underlying mechanism remains elusive. In this study, we investigated the innate immune response to AP-CF. We found that intraperitoneal administration of AP-CF increased the serum level of IL-18 and the number of splenic IFN-c producing CD4+ cells during influenza A virus infection. The adjuvant effect of AP-CF was distinct from that of alum, which is known to have the ability to stimulate a Th2 immune response. In addition, AP-CF injection barely increased the number of peritoneal neutrophils and inflammatory macrophages, whereas alum injection markedly increased the number of neutrophils and inflammatory macrophages, suggesting that AP-CF is a weak inducer of inflammation compared to alum. APCF induced IL-18 production by DC2.4 cells, a dendritic cell line, and by peritoneal exudate cells that include peritoneal macrophages. Collectively, our findings indicate that AP-CF is an adjuvant that promotes the Th1 response during influenza A virus infection.

Aureobasidium pullulans produced β-glucan is effective to enhance Kurosengoku soybean extract induced Thrombospondin-1 expression.
Muramatsu D, Okabe M, Takaoka A, Kida H, Iwai A
Sci Rep, 7, 1, 2831, Nature Publishing Group, 2017年12月01日, ［査読有り］
英語, 研究論文（学術雑誌）, Black yeast, Aureobasidium pullulans is extracellularly produced β-(1,3), (1,6)-D-glucan (β-glucan) under certain conditions. In this study, using Glycine max cv. Kurosengoku (Kurosengoku soybeans), the production of β-glucan through fermentation of A. pullulans was evaluated, and the effects of A. pullulans cultured fluid (AP-CF) containing β-glucan made with Kurosengoku soybeans (kAP-CF) on a human monocyte derived cell line, Mono Mac 6 cells were investigated. Concentration of β-glucan in kAP-CF reached the same level as normal AP-CF. An anti-angiogenic protein, Thrombospondin-1 (THBS1) was effectively induced after the stimulation with kAP-CF for comparison with AP-CF. The THBS1 is also induced after stimulation with hot water extract of Kurosengoku soybeans (KS-E), while the combined stimulation of β-glucan with KS-E more effectively induced THBS1 than that with KS-E alone. These results suggest effects of A. pullulans-produced β-glucan on the enhancement of Kurosengoku soybean-induced THBS1 expression.

A cross-sectional study to quantify the prevalence of avian influenza viruses in poultry at intervention and non-intervention live bird markets in central Vietnam, 2014
D. -H. Chu, M. A. Stevenson, L. V. Nguyen, N. Isoda, S. M. Firestone, T. N. Nguyen, L. T. Nguyen, K. Matsuno, M. Okamatsu, H. Kida, Y. Sakoda
TRANSBOUNDARY AND EMERGING DISEASES, 64, 6, 1991, 1999, WILEY, 2017年12月, ［査読有り］
英語, 研究論文（学術雑誌）, In Vietnam, live bird markets are found in most populated centres, providing the means by which fresh poultry can be purchased by consumers for immediate consumption. Live bird markets are aggregation points for large numbers of poultry, and therefore, it is common for a range of avian influenza viruses to be mixed within live bird markets as a result of different poultry types and species being brought together from different geographical locations. We conducted a cross-sectional study in seven live bird markets in four districts of Thua Thien Hue Province in August and December, 2014. The aims of this study were to (i) document the prevalence of avian influenza in live bird markets (as measured by virus isolation); and (ii) quantify individual bird-, seller- and market-level characteristics that rendered poultry more likely to be positive for avian influenza virus at the time of sale. A questionnaire soliciting details of knowledge, attitude and avian influenza practices was administered to poultry sellers in study markets. At the same time, swabs and faecal samples were collected from individual poultry and submitted for isolation of avian influenza virus. The final data set comprised samples from 1,629 birds from 83 sellers in the seven live bird markets. A total of 113 birds were positive for virus isolation; a prevalence of 6.9 (95% CI 5.8-8.3) avian influenza virus-positive birds per 100 birds submitted for sale. After adjusting for clustering at the market and individual seller levels, none of the explanatory variables solicited in the questionnaire were significantly associated with avian influenza virus isolation positivity. The proportions of variance at the individual market, seller and individual bird levels were 6%, 48% and 46%, respectively. We conclude that the emphasis of avian influenza control efforts in Vietnam should be at the individual seller level as opposed to the market level.

Selection of antigenic variants of an H5N1 highly pathogenic avian influenza virus in vaccinated chickens
Lam Thanh Nguyen, Tatsuya Nishi, Shintaro Shichinohe, Duc-Huy Chu, Takahiro Hiono, Keita Matsuno, Masatoshi Okamatsu, Hiroshi Kida, Yoshihiro Sakoda
VIROLOGY, 510, 252, 261, ACADEMIC PRESS INC ELSEVIER SCIENCE, 2017年10月, ［査読有り］
英語, 研究論文（学術雑誌）, Vaccination-primed immunity in poultry has been suggested for selection of antigenically drifted highly pathogenic avian influenza viruses (HPAIVs). In this study, we performed two consecutive passage studies of an H5N1 HPAIV in vaccinated chickens, namely, study-I and study-II, to select antigenic variants under immune pressure from the vaccination. In study-I, nine consecutive passages of a wild-type H5N1 HPAIV were carried out in chickens vaccinated with the homologous challenge strain. Antigenically drifted variants with mutations at position 179 in the hemagglutinin (HA) were selected after three passages. Similarly, in study-II, a vaccination-mediated antigenic variant isolated in study-I was used as the vaccine and challenge strain to confirm further antigenic drift after updating the vaccine; after the third passage, additional antigenic variants with a mutation at position 256 in the HA were selected. Thus, our study demonstrated the contribution of vaccination in the selection of antigenic variants of H5 HPAIVs in chickens.

Efficacy of recombinant measles virus expressing highly pathogenic avian influenza virus (HPAIV) antigen against HPAIV infection in monkeys
Tomoko Fujiyuki, Ryo Horie, Misako Yoneda, Takeshi Kuraishi, Fumihiko Yasui, Hyun-jeong Kwon, Keisuke Munekata, Fusako Ikeda, Miho Hoshi, Yuri Kiso, Mio Omi, Hiroki Sato, Hiroshi Kida, Shosaku Hattori, Michinori Kohara, Chieko Kai
SCIENTIFIC REPORTS, 7, 1, 12017, NATURE PUBLISHING GROUP, 2017年09月, ［査読有り］
英語, 研究論文（学術雑誌）, Highly pathogenic avian influenza virus (HPAIV) is a serious threat not only to domestic fowls but also to humans. Vaccines inducing long-lasting immunity against HPAIV are required. In the present study, we generated recombinant measles virus (MV) expressing the hemagglutinin protein of HPAIV without the multibasic site necessary for its pathogenicity in chickens using the backbone of an MV vaccine strain (rMV-Ed-H5HA) or a wild-type MV-derived mutant (rMV-HL-Vko-H5HA). We examined protective efficacy of the candidate vaccines in the monkey infection model by the challenge with a HPAIV (H5N1). Cynomolgus monkeys inoculated with the candidate vaccines produced both anti-H5 HA and anti-MV antibodies. They recovered earlier from influenza symptoms than unvaccinated monkeys after the challenge with the HPAIV strain. Chest radiography and histopathological analyses confirmed less severe pneumonia in the vaccinated monkeys. Vaccination tended to suppress viral shedding and reduced the interleukin-6 levels in the lungs. Furthermore, the vaccination with rMV-Ed-H5HA of monkeys with pre-existing anti-MV immunity induced the production of anti-H5 HA antibodies. These results suggest that both candidate vaccines effectively reduce disease severity in naive hosts, and that rMV-Ed-H5HA is a particularly good candidate vaccine against HPAIV infection.

Characterization of H5N6 highly pathogenic avian influenza viruses isolated from wild and captive birds in the winter season of 2016-2017 in Northern Japan
Takahiro Hiono, Masatoshi Okamatsu, Keita Matsuno, Atsushi Haga, Ritsuko Iwata, Lam Thanh Nguyen, Mizuho Suzuki, Yuto Kikutani, Hiroshi Kida, Manabu Onuma, Yoshihiro Sakoda
MICROBIOLOGY AND IMMUNOLOGY, 61, 9, 387, 397, WILEY, 2017年09月, ［査読有り］
英語, 研究論文（学術雑誌）, On 15 November 2016, a black swan that had died in a zoo in Akita prefecture, northern Japan, was strongly suspected to have highly pathogenic avian influenza (HPAI); an HPAI virus (HPAIV) belonging to the H5N6 subtype was isolated from specimens taken from the bird. After the initial report, 230 cases of HPAI caused by H5N6 viruses from wild birds, captive birds, and domestic poultry farms were reported throughout the country during the winter season. In the present study, 66 H5N6 HPAIVs isolated from northern Japan were further characterized. Phylogenetic analysis of the hemagglutinin gene showed that the H5N6 viruses isolated in northern Japan clustered into Group C of Clade 2.3.4.4 together with other isolates collected in Japan, Korea and Taiwan during the winter season of 2016-2017. The antigenicity of the Japanese H5N6 isolate differed slightly from that of HPAIVs isolated previously in Japan and China. The virus exhibited high pathogenicity and a high replication capacity in chickens, whereas virus growth was slightly lower in ducks compared with that of an H5N8 HPAIV isolate collected in Japan in 2014. Comprehensive analyses of Japanese isolates, including those from central, western, and southern Japan, as well as rapid publication of this information are essential for facilitating greater control of HPAIVs.

Rapid and broad detection of H5 hemagglutinin by an immunochromatographic kit using novel monoclonal antibody against highly pathogenic avian influenza virus belonging to the genetic clade 2.3.4.4
Lam Thanh Nguyen, Kazunari Nakaishi, Keiko Motojima, Ayako Ohkawara, Erina Minato, Junki Maruyama, Takahiro Hiono, Keita Matsuno, Masatoshi Okamatsu, Takashi Kimura, Ayato Takada, Hiroshi Kida, Yoshihiro Sakoda
PLOS ONE, 12, 8, e0182228, PUBLIC LIBRARY SCIENCE, 2017年08月, ［査読有り］
英語, 研究論文（学術雑誌）, Highly pathogenic avian influenza viruses (HPAIVs) of H5 subtype have persistently caused outbreaks in domestic poultry and wild birds worldwide and sporadically infected humans. Rapid and accurate diagnosis is one of the key strategies for the control of H5 HPAIVs. However, the sensitivity of the diagnosis of H5 HPAIVs has gradually reduced due to extensive antigenic variation during their evolution. Particularly, the previously developed immunochromatographic diagnosis kit for H5 viruses, Linjudge Flu A/H5, exhibits reduced detection of H5 HPAIVs isolated in recent years. In the present study, we established a new advanced H5 rapid immunochromatographic detection kit (New Linjudge Flu A/H5) by a combination of two anti-H5 hemagglutinin monoclonal antibodies, A64/1 previously applied in the Linjudge Flu A/H5 and A32/2, a novel monoclonal antibody generated from a clade 2.3.4.4 H5 HPAIV. The new kit broadly detected all classical and recent H5 influenza viruses and showed a higher specificity and sensitivity than the original Linjudge Flu A/H5 with recently circulating H5 HPAIVs. Furthermore, the applicability of the New Linjudge Flu A/H5 was demonstrated by detecting antigens from the swabs and tissue homogenates of naturally infected birds and experimentally infected chickens with H5N6 HPAIVs belonging to the genetic clade 2.3.4.4. Our study, therefore, can provide an effective point-of-care rapid antigen detection kit for the surveillance of H5 avian influenza viruses and as a prompt countermeasure against the current widespread of the clade 2.3.4.4 H5 HPAIVs in domestic and wild birds.

Antigenic diversity of H5 highly pathogenic avian influenza viruses of clade 2.3.4.4 isolated in Asia
Ayako Ohkawara, Masatoshi Okamatsu, Makoto Ozawa, Duc-Huy Chu, Lam Thanh Nguyen, Takahiro Hiono, Keita Matsuno, Hiroshi Kida, Yoshihiro Sakoda
MICROBIOLOGY AND IMMUNOLOGY, 61, 5, 149, 158, WILEY, 2017年05月, ［査読有り］
英語, 研究論文（学術雑誌）, H5 highly pathogenic avian influenza viruses (HPAIV) have spread in both poultry and wild birds since late 2003. Continued circulation of HPAIV in poultry in several regions of the world has led to antigenic drift. In the present study, we analyzed the antigenic properties of H5 HPAIV isolated in Asia using four neutralizing mAbs recognizing hemagglutinin, which were established using A/chicken/Kumamoto/1-7/2014 (H5N8), belonging to clade 2.3.4.4 and also using polyclonal antibodies. Viruses of clades 1.1, 2.3.2.1, 2.3.4, and 2.3.4.4 had different reactivity patterns to the panel of mAbs, thereby indicating that the antigenicity of the viruses of clade 2.3.4.4 were similar but differed from the other clades. In particular, the antigenicity of the viruses of clade 2.3.4.4 differed from those of the viruses of clades 2.3.4 and 2.3.2.1, which suggests that the recent H5 HPAIV have further evolved antigenically divergent. In addition, reactivity of antiserum suggests that the antigenicity of viruses of clade 2.3.4.4 differed slightly among groups A, B, and C. Vaccines are still used in poultry in endemic countries, so the antigenicity of H5 HPAIV should be monitored continually to facilitate control of avian influenza. The panel of mAbs established in the present study will be useful for detecting antigenic drift in the H5 viruses that emerge from the current strains.

Potency of whole virus particle and split virion vaccines using dissolving microneedle against challenges of H1N1 and H5N1 influenza viruses in mice
Akihiro Nakatsukasa, Koji Kuruma, Masatoshi Okamatsu, Takahiro Hiono, Mizuho Suzuki, Keita Matsuno, Hiroshi Kida, Takayoshi Oyamada, Yoshihiro Sakoda
VACCINE, 35, 21, 2855, 2861, ELSEVIER SCI LTD, 2017年05月, ［査読有り］
英語, 研究論文（学術雑誌）, Transdermal vaccination using a microneedle (MN) confers enhanced immunity compared with subcutaneous (SC) vaccination. Here we developed a novel dissolving MN patch for the influenza vaccine. The potencies of split virion and whole virus particle (WVP) vaccines prepared from A/Puerto Rico/8/1934 (H1N1) and A/duck/Hokkaido/Vac-3/2007 (H5N1), respectively, were evaluated. MN vaccination induced higher neutralizing antibody responses than SC vaccination in mice. Moreover, MN vaccination with a lower dose of antigens conferred protective immunity against lethal challenges of influenza viruses than SC vaccination in mice. These results suggest that the WVP vaccines administered using MN are an effective combination for influenza vaccine to be further validated in humans. (C) 2017 Elsevier Ltd. All rights reserved.

Characterization of Highly Pathogenic Avian Influenza Virus A(H5N6), Japan, November 2016
Masatoshi Okamatsu, Makoto Ozawa, Kosuke Soda, Hiroki Takakuwa, Atsushi Haga, Takahiro Hiono, Aya Matsuu, Yuko Uchida, Ritsuko Iwata, Keita Matsuno, Masakazu Kuwahara, Toshiyo Yabuta, Tatsufumi Usui, Hiroshi Ito, Manabu Onuma, Yoshihiro Sakoda, Takehiko Saito, Koichi Otsuki, Toshihiro Ito, Hiroshi Kida
EMERGING INFECTIOUS DISEASES, 23, 4, 691, 695, CENTERS DISEASE CONTROL & PREVENTION, 2017年04月, ［査読有り］
英語, 研究論文（学術雑誌）, Highly pathogenic avian influenza viruses (HPAIVs) A(H5N6) were concurrently introduced into several distant regions of Japan in November 2016. These viruses were classified into the genetic clade 2.3.4.4c and were genetically closely related to H5N6 HPAIVs recently isolated in South Korea and China. In addition, these HPAIVs showed further antigenic drift.

Therapeutic efficacy of peramivir against H5N1 highly pathogenic avian influenza viruses harboring the neuraminidase H275Y mutation
Masanori Kobayashi, Makoto Kodama, Takeshi Noshi, Ryu Yoshida, Takushi Kanazu, Naoki Nomura, Kosuke Soda, Norikazu Isoda, Masatoshi Okamatsu, Yoshihiro Sakoda, Yoshinori Yamano, Akihiko Sato, Hiroshi Kida
ANTIVIRAL RESEARCH, 139, 41, 48, ELSEVIER SCIENCE BV, 2017年03月, ［査読有り］
英語, 研究論文（学術雑誌）, High morbidity and mortality associated with human cases of highly pathogenic avian influenza (HPAI) viruses, including H5N1 influenza virus, have been reported. The purpose of the present study was to evaluate the antiviral effects of peramivir against HPAI viruses. In neuraminidase (NA) inhibition and virus replication inhibition assays, peramivir showed strong inhibitory activity against H5N1, H7N1 and H7N7 HPAI viruses with sub-nanomolar activity in enzyme assays. In H5N1 viruses containing the NA H275Y mutation, the antiviral activity of peramivir against the variant was lower than that against the wild-type. Evaluation of the in vivo antiviral activity showed that a single intravenous treatment of peramivir (10 mg/kg) prevented lethality in mice infected with wild-type H5N1 virus and also following infection with H5N1 virus with the H275Y mutation after a 5 day administration of peramivir (30 mg/kg). Furthermore, mice injected with peramivir showed low viral titers and low levels of proinflammatory cytokines in the lungs. These results suggest that peramivir has therapeutic activity against HPAI viruses even if the virus harbors the NA H275Y mutation. (C) 2016 Elsevier B.V. All rights reserved.

Potential risk of repeated nasal vaccination that induces allergic reaction with mucosal IgE and airway eosinophilic infiltration in cynomolgus macaques infected with H5N1 highly pathogenic avian influenza virus
Misako Nakayama, Yasushi Itoh, Shintaro Shichinohe, Rumi Nakabayashi, Hirohito Ishigaki, Yoshihiro Sakoda, Quynh Mai Le, Yoshihiro Kawaoka, Hiroshi Kida, Kazumasa Ogasawara
VACCINE, 35, 7, 1008, 1017, ELSEVIER SCI LTD, 2017年02月, ［査読有り］
英語, 研究論文（学術雑誌）, The efficacy and detrimental effect of mucosal vaccination with an inactivated influenza vaccine were examined in a macaque model by intranasal administration with small amounts of inactivated whole virus particles and challenge by a human-derived H5N1 highly pathogenic avian influenza virus infection. Repeated nasal inoculation with the whole particle vaccine of an inactivated virus, A/duck/Hokkaido/Vac-3/2007 (H5N1) (Vac-3), induced antigen-specific IgA and IgG antibody production in nasal swabs and plasma. Vac-3-specific IgE production was also found in the nasal swabs. Nasal vaccination with Vac-3 induced broader cross-clade neutralization activity than did subcutaneous vaccination. After challenge infection, repeated nasal vaccination almost completely prevented the propagation of virus in the upper and lower airways and protected cynomolgus macaques from viral pneumonia by induction of IgA-producing B cells in the lungs. On the other hand, eosinophil clusters were observed in the lungs of vaccinated macaques. Although Vac-3-specific IgE antibody and IL-13 levels were decreased after infection compared to those before infection and no anaphylaxis in vaccinated macaques was detected after challenge infection, our results suggest that we have to pay attention to potential allergic responses at repeated nasal vaccination, especially in people who have an airway allergy. (C) 2017 Elsevier Ltd. All rights reserved.

Vaccination against H9N2 avian influenza virus reduces bronchus-associated lymphoid tissue formation in cynomolgus macaques after intranasal virus challenge infection
Misako Nakayama, Hiroichi Ozaki, Yasushi Itoh, Kosuke Soda, Hirohito Ishigaki, Masatoshi Okamatsu, Yoshihiro Sakoda, Chun-Ho Park, Hideaki Tsuchiya, Hiroshi Kida, Kazumasa Ogasawara
PATHOLOGY INTERNATIONAL, 66, 12, 678, 686, WILEY-BLACKWELL, 2016年12月, ［査読有り］
英語, 研究論文（学術雑誌）, H9N2 avian influenza virus causes sporadic human infection. Since humans do not possess acquired immunity specific to this virus, we examined the pathogenicity of an H9N2 virus isolated from a human and then analyzed protective effects of a vaccine in cynomolgus macaques. After intranasal challenge with A/Hong Kong/1073/1999 (H9N2) (HK1073) isolated from a human patient, viruses were isolated from nasal and tracheal swabs in unvaccinated macaques with mild fever and body weight loss. A formalin-inactivated H9N2 whole particle vaccine derived from our virus library was subcutaneously inoculated to macaques. Vaccination induced viral antigen-specific IgG and neutralization activity in sera. After intranasal challenge with H9N2, the virus was detected only the day after inoculation in the vaccinated macaques. Without vaccination, many bronchus-associated lymphoid tissues (BALTs) were formed in the lungs after infection, whereas the numbers of BALTs were smaller and the cytokine responses were weaker in the vaccinated macaques than those in the unvaccinated macaques. These findings indicate that the H9N2 avian influenza virus HK1073 is pathogenic in primates but seems to cause milder symptoms than does H7N9 influenza virus as found in our previous studies and that a formalin-inactivated H9N2 whole particle vaccine induces protective immunity against H9N2 virus.

Sensitization with vaccinia virus encoding H5N1 hemagglutinin restores immune potential against H5N1 influenza virus
Fumihiko Yasui, Yasushi Itoh, Ai Ikejiri, Masahiro Kitabatake, Nobuo Sakaguchi, Keisuke Munekata, Shintaro Shichinohe, Yukiko Hayashi, Hirohito Ishigaki, Misako Nakayama, Yoshihiro Sakoda, Hiroshi Kida, Kazumasa Ogasawara, Michinori Kohara
SCIENTIFIC REPORTS, 6, 37915, NATURE PUBLISHING GROUP, 2016年11月, ［査読有り］
英語, 研究論文（学術雑誌）, H5N1 highly pathogenic avian influenza (H5N1 HPAI) virus causes elevated mortality compared with seasonal influenza viruses like H1N1 pandemic influenza (H1N1 pdm) virus. We identified a mechanism associated with the severe symptoms seen with H5N1 HPAI virus infection. H5N1 HPAI virus infection induced a decrease of dendritic cell number in the splenic extrafollicular T-cell zone and impaired formation of the outer layers of B-cell follicles, resulting in insufficient levels of antibody production after infection. However, in animals vaccinated with a live recombinant vaccinia virus expressing the H5 hemagglutinin, infection with H5N1 HPAI virus induced parafollicular dendritic cell accumulation and efficient antibody production. These results indicate that a recombinant vaccinia encoding H5 hemagglutinin gene does not impair dendritic cell recruitment and can be a useful vaccine candidate.

Role for migratory wild birds in the global spread of avian influenza H5N8
Samantha J. Lycett, Rogier Bodewes, Anne Pohlmann, Jill Banks, Krisztian Banyai, Maciej F. Boni, Ruth Bouwstra, Andrew C. Breed, Ian H. Brown, Hualan Chen, Adam Dan, Thomas J. DeLiberto, Nguyen Diep, Marius Gilbert, Sarah Hill, Hon S. Ip, Chang Wen Ke, Hiroshi Kida, Mary Lea Killian, Marion P. Koopmans, Jung-Hoon Kwon, Dong-Hun Lee, Youn Jeong Lee, Lu Lu, Isabella Monne, John Pasick, Oliver G. Pybus, Andrew Rambaut, Timothy P. Robinson, Yoshihiro Sakoda, Siamak Zohari, Chang-Seon Song, David E. Swayne, Mia Kim Torchetti, Hsiang-Jung Tsai, Ron A. M. Fouchier, Martin Beer, Mark Woolhouse, Thijs Kuiken
SCIENCE, 354, 6309, 213, 217, AMER ASSOC ADVANCEMENT SCIENCE, 2016年10月, ［査読有り］
英語, 研究論文（学術雑誌）, Avian influenza viruses affect both poultry production and public health. A subtype H5N8 (clade 2.3.4.4) virus, following an outbreak in poultry in South Korea in January 2014, rapidly spread worldwide in 2014-2015. Our analysis of H5N8 viral sequences, epidemiological investigations, waterfowl migration, and poultry trade showed that long-distance migratory birds can play a major role in the global spread of avian influenza viruses. Further, we found that the hemagglutinin of clade 2.3.4.4 virus was remarkably promiscuous, creating reassortants with multiple neuraminidase subtypes. Improving our understanding of the circumpolar circulation of avian influenza viruses in migratory waterfowl will help to provide early warning of threats from avian influenza to poultry, and potentially human, health.

Recent developments in the diagnosis of avian influenza
Masatoshi Okamatsu, Takahiro Hiono, Hiroshi Kida, Yoshihiro Sakoda
VETERINARY JOURNAL, 215, 82, 86, ELSEVIER SCI LTD, 2016年09月, ［査読有り］
英語, The diagnosis of influenza A virus infections in poultry or wild birds is difficult due to variations in the pathogenicity of the viruses in different avian hosts and also the antigenic and genetic diversity of the virus, particularly the recent H5 highly pathogenic avian influenza viruses. A classical standard laboratory technique is virus isolation prior to subtyping and pathotyping. This diagnostic technique is crucial for further virological analyses, particularly during an initial outbreak; however, delays in diagnosis have thwarted effective disease control in recent years. Recent developments in molecular biological techniques provide an accelerated diagnosis. Such technologies, which include real-time reverse transcriptase PCR, isothermal nucleic acid amplification, next-generation sequencing and immunochromatography, contribute to simpler and more rapid diagnosis. The advantages of each of these diagnostic techniques should be considered for effective control of avian influenza. (C) 2016 Elsevier Ltd. All rights reserved.

Is the optimal pH for membrane fusion in host cells by avian influenza viruses related to host range and pathogenicity?
Masatoshi Okamatsu, Yurie Motohashi, Takahiro Hiono, Tomokazu Tamura, Kazuki Nagaya, Keita Matsuno, Yoshihiro Sakoda, Hiroshi Kida
ARCHIVES OF VIROLOGY, 161, 8, 2235, 2242, SPRINGER WIEN, 2016年08月, ［査読有り］
英語, 研究論文（学術雑誌）, Influenza viruses isolated from wild ducks do not replicate in chickens. This fact is not explained solely by the receptor specificity of the hemagglutinin (HA) from such viruses for target host cells. To investigate this restriction in host range, the fusion activities of HA molecules from duck and chicken influenza viruses were examined. Influenza viruses A/duck/Mongolia/54/2001 (H5N2) (Dk/MNG) and A/chicken/Ibaraki/1/2005 (H5N2) (Ck/IBR), which replicate only in their primary hosts, were used. The optimal pH for membrane fusion of Ck/IBR was 5.9, higher than that of Dk/MNG at 4.9. To assess the relationship between the optimal pH for fusion and the host range of avian influenza viruses, the optimal pH for fusion of 55 influenza virus strains isolated from ducks and chickens was examined. No correlation was found between the host range and optimal pH for membrane fusion by the viruses, and this finding applied also to the H5N1 highly pathogenic avian influenza viruses. The optimal pH for membrane fusion for avian influenza viruses was shown to not necessarily be correlated with their host range or pathogenicity in ducks and chickens.

Genetic and antigenic characterization of H5, H6 and H9 avian influenza viruses circulating in live bird markets with intervention in the center part of Vietnam.
Chu DH, Okamatsu M, Matsuno K, Hiono T, Ogasawara K, Nguyen LT, Van Nguyen L, Nguyen TN, Nguyen TT, Van Pham D, Nguyen DH, Nguyen TD, To TL, Van Nguyen H, Kida H, Sakoda Y
Vet Microbiol, 192, 194, 203, 2016年07月, ［査読有り］

Cytokine responses to eye spray adjuvants for enhancing vaccine-induced immunity in chickens.
Hiromi Takaki, Haruko Sato, Riho Kurata, Hirokazu Hikono, Takahiro Hiono, Hiroshi Kida, Misako Matsumoto, Takehiko Saito, Tsukasa Seya
Microbiology and immunology, 60, 7, 511, 5, 2016年07月, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, Eye spray influenza vaccines for chickens are increasingly available; however, how to enhance cellular and antibody responses to them remains undetermined. Here, eye-drops containing the immune-enhancing adjuvants Pam2CSK4 or polyI:C were assessed in chickens. Application of these TLR agonists to chicken conjunctiva resulted in up-regulation of IL-1β, but not other cytokines, including IFN and IL-6, in the spleen, lung and Harderian gland. Thus, responses to adjuvant applied to the conjunctival mucosa of chickens differ from those expected from the responses to intra-nasal adjuvants in mammals. Identifying an appropriate delivery route for adjuvants is crucial for evoking immune responses in chickens.

Constitutive aryl hydrocarbon receptor signaling constrains type I interferon-mediated antiviral innate defense
Taisho Yamada, Hiromasa Horimoto, Takeshi Kameyama, Sumio Hayakawa, Hiroaki Yamato, Masayoshi Dazai, Ayato Takada, Hiroshi Kida, Debbie Bott, Angela C. Zhou, David Hutin, Tania H. Watts, Masahiro Asaka, Jason Matthews, Akinori Takaoka
NATURE IMMUNOLOGY, 17, 6, 687, +, NATURE PUBLISHING GROUP, 2016年06月, ［査読有り］
英語, 研究論文（学術雑誌）, Aryl hydrocarbon receptor (AHR) is a ligand-activated transcription factor that mediates the toxic activity of many environmental xenobiotics. However, its role in innate immune responses during viral infection is not fully understood. Here we demonstrate that constitutive AHR signaling negatively regulates the type I interferon (IFN-I) response during infection with various types of virus. Virus-induced IFN-beta production was enhanced in AHR-deficient cells and mice and resulted in restricted viral replication. We found that AHR upregulates expression of the ADP-ribosylase TIPARP, which in turn causes downregulation of the IFN-I response. Mechanistically, TIPARP interacted with the kinase TBK1 and suppressed its activity by ADP-ribosylation. Thus, this study reveals the physiological importance of endogenous activation of AHR signaling in shaping the IFN-I-mediated innate response and, further, suggests that the AHR-TIPARP axis is a potential therapeutic target for enhancing antiviral responses.

Toll-like receptor 9 ligand D-type oligodeoxynucleotide D35 as a broad inhibitor for influenza A virus replication that is associated with suppression of neuraminidase activity
Hiroshi Yamada, Satoshi Nagase, Kazuo Takahashi, Yoshihiro Sakoda, Hiroshi Kida, Shigefumi Okamoto
ANTIVIRAL RESEARCH, 129, 81, 92, ELSEVIER SCIENCE BV, 2016年05月, ［査読有り］
英語, 研究論文（学術雑誌）, The most effective drugs available to treat influenza are neuraminidase (NA) inhibitors, which provide important additional measures for the control of influenza virus infections. However, since the emergence of NA inhibitor-resistant viruses may compromise the clinical utility of this class of anti-influenza agents, it is very important to develop new anti-influenza agents which target a different region in NA responsible for its sensitivity from that for NA inhibitors and could be used to treat NA inhibitors resistant isolates. The oligodeoxynucleotide D35, multimerized and aggregated, suppressed replication of influenza A viruses except A/WSN/33 (WSN). The suppressive viral replication by D35 depended on G-terad and multimer formation. The range of the suppressive viral replication at the late stage, including virus assembly and release from infected cells, was much larger than that at the initial stage, viral attachment and entry. D35 suppressed NA activity of influenza A viruses. Furthermore, replacing the NA gene of A/Puerto Rico/8/34 (PR8), in which viral replication was inhibited by D35 at the late stage, with the NA gene from WSN, in which viral replication was not inhibited, eliminated the D35-dependent suppression. D35 showed an additive anti-influenza effect with oseltamivir. It was also effective in vivo. These results suggest that the influenza virus NA mainly contributse to the D35-suppressible virus release from infected cells at the late stage. In addition, because administration of D35 into the virus-infected mice suppressed viral replication and weight loss, clinical application of D35 could be considered. (C) 2016 Elsevier B.V. All rights reserved.

Comparison of pathogenicities of H7 avian influenza viruses via intranasal and conjunctival inoculation in cynomolgus macaques.
Shichinohe S, Itoh Y, Nakayama M, Ozaki H, Soda K, Ishigaki H, Okamatsu M, Sakoda Y, Kida H
Virology, 493, 31, 38, 2016年03月, ［査読有り］
研究論文（学術雑誌）

Amino acid residues at positions 222 and 227 of the hemagglutinin together with the neuraminidase determine binding of H5 avian influenza viruses to sialyl Lewis X
Takahiro Hiono, Masatoshi Okamatsu, Manabu Igarashi, Ryan McBride, Robert P. de Vries, Wenjie Peng, James C. Paulson, Yoshihiro Sakoda, Hiroshi Kida
ARCHIVES OF VIROLOGY, 161, 2, 307, 316, SPRINGER WIEN, 2016年02月, ［査読有り］
英語, 研究論文（学術雑誌）, Influenza viruses isolated from ducks are rarely able to infect chickens; it is therefore postulated that these viruses need to adapt in some way to be able to be transmitted to chickens in nature. Previous studies revealed that sialyl Lewis X (3'SLeX), which is fucosylated alpha 2,3 sialoside, was predominantly detected on the epithelial cells of the chicken trachea, whereas this glycan structure is not found in the duck intestinal tract. To clarify the mechanisms of the interspecies transmission of influenza viruses between ducks and chickens, we compared the receptor specificity of low-pathogenic avian influenza viruses isolated from these two species. Glycan-binding analysis of the recombinant hemagglutinin (HA) of a chicken influenza virus, A/chicken/Ibaraki/1/2005 (H5N2), revealed a binding preference to alpha 1,3 fucosylated sialosides. On the other hand, the HA of a duck influenza virus, A/duck/Mongolia/54/2001 (H5N2) (Dk/MNG), particularly bound to non-fucosylated alpha 2,3 sialosides such as 3'-sialyllactosamine (3'SLacNAc). Computational analysis along with binding analysis of the mutant HAs revealed that this glycan-binding specificity of the HA was determined by amino acid residues at positions 222 and 227. Inconsistent with the glycan-binding specificity of the recombinant HA protein, virions of Dk/MNG bound to both 3'SLacNAc and 3'SLeX. Glycan-binding analysis in the presence of a neuraminidase (NA) inhibitor revealed that the NA conferred binding to 3'SLeX to virions of Dk/MNG. The present results reveal the molecular basis of the interaction between fucosylated alpha 2,3 sialosides and influenza viruses.

Experimental infection of highly and low pathogenic avian influenza viruses to chicken's, ducks, tree sparrows, jungle crows, and black rats for the evaluation of their roles in virus transmission
Takahiro Hiono, Masatoshi Okamatsu, Naoki Yamamoto, Kohei Ogasawara, Mayumi Endo, Saya Kuribayashi, Shintaro Shichinohe, Yurie Motohashi, Duc-Huy Chu, Mizuho Suzuki, Takaya Ichikawa, Tatsuya Nishi, Yuri Abe, Keita Matsuno, Kazuyuki Tanaka, Tsutomu Tanigawa, Hiroshi Kida, Yoshihiro Sakoda
VETERINARY MICROBIOLOGY, 182, 108, 115, ELSEVIER SCIENCE BV, 2016年01月, ［査読有り］
英語, 研究論文（学術雑誌）, Highly pathogenic avian influenza viruses (HPAIVs) have spread in both poultry and wild birds. Determining transmission routes of these viruses during an outbreak is essential for the control of avian influenza. It has been widely postulated that migratory ducks play crucial roles in the widespread dissemination of HPAIVs in poultry by carrying viruses along with their migrations; however close contacts between wild migratory ducks and poultry are less likely in modern industrial poultry farming settings. Therefore, we conducted experimental infections of HPAIVs and low pathogenic avian influenza viruses (LPAIVs) to chickens, domestic ducks, tree sparrows, jungle crows, and black rats to evaluate their roles in virus transmission. The results showed that chickens, ducks, sparrows, and crows were highly susceptible to HPAIV infection. Significant titers of virus were recovered from the sparrows and crows infected with HPAIVs, which suggests that they potentially play roles of transmission of HPAIVs to poultry. In contrast, the growth of LPAIVs was limited in each of the animals tested compared with that of HPAIVs. The present results indicate that these common synanthropes play some roles in influenza virus transmission from wild birds to poultry. (C) 2015 Elsevier B.V. All rights reserved.

Protective efficacy of stockpiled vaccine against H5N8 highly pathogenic avian influenza virus isolated from a chicken in Kumamoto prefecture, Japan, in 2014
Koichiro Gamoh, Mari Nakamizo, Masatoshi Okamatsu, Yoshihiro Sakoda, Hiroshi Kida, Shoko Suzuki
JOURNAL OF VETERINARY MEDICAL SCIENCE, 78, 1, 139, 142, JAPAN SOC VET SCI, 2016年01月, ［査読有り］
英語, 研究論文（学術雑誌）, H5 highly pathogenic avian influenza (HPAI) viruses have spread worldwide, and antigenic variants of different clades have been selected. In this study, the national stockpiled vaccine prepared from A/duck/HoldcaidoNac-1/2004 (H5N1) strain was evaluated for the protective efficacy against H5N8 HPAI virus isolated in Kumamoto prefecture, Japan, in April 2014. In the challenge test, all of the vaccinated chickens survived without showing any clinical signs and reduced virus shedding. It was concluded that the present stockpiled vaccine was effective against the H5N8 HPAI virus.

Genetic and antigenic characterization of bovine viral diarrhea viruses isolated from cattle in Hokkaido, Japan
Yuri Abe, Tomokazu Tamura, Shiho Torii, Shiho Wakamori, Makoto Nagai, Kazuya Mitsuhashi, Junki Mine, Yuri Fujimoto, Naofumi Nagashima, Fumi Yoshino, Yukihiko Sugita, Takushi Nomura, Masatoshi Okamatsu, Hiroshi Kida, Yoshihiro Sakoda
JOURNAL OF VETERINARY MEDICAL SCIENCE, 78, 1, 61, 70, JAPAN SOC VET SCI, 2016年01月, ［査読有り］
英語, 研究論文（学術雑誌）, In our previous study, we genetically analyzed bovine viral diarrhea viruses (BVDVs) isolated from 2000 to 2006 in Japan and reported that subgenotype 1b viruses were predominant. In the present study, 766 BVDVs isolated from 2006 to 2014 in Hokkaido, Japan, were genetically analyzed to understand recent epidemics. Phylogenetic analysis based on nucleotide sequences of the 5'-untranslated region of viral genome revealed that 766 isolates were classified as genotype 1 (BVDV-1; 544 isolates) and genotype 2 (BVDV-2; 222). BVDV-1 isolates were further divided into BVDV-la (93), 1b (371) and 1c (80) subgenotypes, and all BVDV-2 isolates were grouped into BVDV-2a subgenotype (222). Further comparative analysis was performed with BVDV-1a, 1b and 2a viruses isolated from 2001 to 2014. Phylogenetic analysis based on nucleotide sequences of the viral glycoprotein E2 gene, a major target of neutralizing antibodies, revealed that BVDV-1a, 1b and 2a isolates were further classified into several clusters. Cross-neutralization tests showed that BVDV-1b isolates were antigenically different from BVDV-1a isolates, and almost BVDV-1a, 1b and 2a isolates were antigenically similar among each subgenotype and each E2 cluster. Taken together, BVDV-1b viruses are still predominant, and BVDV-2a viruses have increased recently in Hokkaido, Japan. Field isolates of BVDV-1a, 1b and 2a show genetic diversity on the E2 gene with antigenic conservation among each subgenotype during the last 14 years.

豚コレラウイルス非構造蛋白NS4BのN末端領域の機能解析　　　　　　　　　　　　　　　
田村友和, Nicolas Ruggli, 長島尚史, 岡松正敏, Artur Summerfield, 松野啓太, 喜田宏, 迫田義博
2015年09月, ［査読有り］
日本語, 研究論文（その他学術会議資料等）

Intracellular membrane association of the N-terminal domain of classical swine fever virus NS4B determines viral genome replication and virulence
Tomokazu Tamura, Nicolas Ruggli, Naofumi Nagashima, Masatoshi Okamatsu, Manabu Igarashi, Junki Mine, Martin A. Hofmann, Matthias Liniger, Artur Summerfield, Hiroshi Kida, Yoshihiro Sakoda
JOURNAL OF GENERAL VIROLOGY, 96, 9, 2623, 2635, SOC GENERAL MICROBIOLOGY, 2015年09月, ［査読有り］
英語, 研究論文（学術雑誌）, Classical swine fever virus (CSFV) causes a highly contagious disease in pigs that can range from a severe haemorrhagic fever to a nearly unapparent disease, depending on the virulence of the virus strain. Little is known about the viral molecular determinants of CSFV virulence. The nonstructural protein NS4B is essential for viral replication. However, the roles of CSFV NS4B in viral genome replication and pathogenesis have not yet been elucidated. NS4B of the GPE(-) vaccine strain and of the highly virulent Eystrup strain differ by a total of seven amino acid residues, two of which are located in the predicted trans-membrane domains of NS4B and were described previously to relate to virulence, and five residues clustering in the N-terminal part. In the present study, we examined the potential role of these five amino acids in modulating genome replication and determining pathogenicity in pigs. A chimeric low virulent GPE(-)-derived virus carrying the complete Eystrup NS4B showed enhanced pathogenicity in pigs. The in vitro replication efficiency of the NS4B chimeric GPE- replicon was significantly higher than that of the replicon carrying only the two Eystrup-specific amino acids in NS4B. In silico and in vitro data suggest that the N-terminal part of NS4B forms an amphipathic a-helix structure. The N-terminal NS4B with these five amino acid residues is associated with the intracellular membranes. Taken together, this is the first gain-of-function study showing that the N-terminal domain of NS4B can determine CSFV genome replication in cell culture and viral pathogenicity in pigs.

A Single Amino Acid in the M1 Protein Responsible for the Different Pathogenic Potentials of H5N1 Highly Pathogenic Avian Influenza Virus Strains
Naganori Nao, Masahiro Kajihara, Rashid Manzoor, Junki Maruyama, Reiko Yoshida, Mieko Muramatsu, Hiroko Miyamoto, Manabu Igarashi, Nao Eguchi, Masahiro Sato, Tatsunari Kondoh, Masatoshi Okamatsu, Yoshihiro Sakoda, Hiroshi Kida, Ayato Takada
PLOS ONE, 10, 9, e0137989, PUBLIC LIBRARY SCIENCE, 2015年09月, ［査読有り］
英語, 研究論文（学術雑誌）, Two highly pathogenic avian influenza virus strains, A/duck/Hokkaido/WZ83/2010 ( H5N1) (WZ83) and A/duck/Hokkaido/WZ101/2010 (H5N1) (WZ101), which were isolated from wild ducks in Japan, were found to be genetically similar, with only two amino acid differences in their M1 and PB1 proteins at positions 43 and 317, respectively. We found that both WZ83 and WZ101 caused lethal infection in chickens but WZ101 killed them more rapidly than WZ83. Interestingly, ducks experimentally infected with WZ83 showed no or only mild clinical symptoms, whereas WZ101 was highly lethal. We then generated reassortants between these viruses and found that exchange of the M gene segment completely switched the pathogenic phenotype in both chickens and ducks, indicating that the difference in the pathogenicity for these avian species between WZ83 and WZ101 was determined by only a single amino acid in the M1 protein. It was also found that WZ101 showed higher pathogenicity than WZ83 in mice and that WZ83, whose M gene was replaced with that of WZ101, showed higher pathogenicity than wild-type WZ83, although this reassortant virus was not fully pathogenic compared to wild-type WZ101. These results suggest that the amino acid at position 43 of the M1 protein is one of the factors contributing to the pathogenicity of H5N1 highly pathogenic avian influenza viruses in both avian and mammalian hosts.

Genetic and antigenic characterization of H5 and H7 influenza viruses isolated from migratory water birds in Hokkaido, Japan and Mongolia from 2010 to 2014
Takahiro Hiono, Ayako Ohkawara, Kohei Ogasawara, Masatoshi Okamatsu, Tomokazu Tamura, Duc-Huy Chu, Mizuho Suzuki, Saya Kuribayashi, Shintaro Shichinohe, Ayato Takada, Hirohito Ogawa, Reiko Yoshida, Hiroko Miyamoto, Naganori Nao, Wakako Furuyama, Junki Maruyama, Nao Eguchi, Gerelmaa Ulziibat, Bazarragchaa Enkhbold, Munkhduuren Shatar, Tserenjav Jargalsaikhan, Selenge Byambadorj, Batchuluun Damdinjav, Yoshihiro Sakoda, Hiroshi Kida
VIRUS GENES, 51, 1, 57, 68, SPRINGER, 2015年08月, ［査読有り］
英語, 研究論文（学術雑誌）, Migratory water birds are the natural reservoir of influenza A viruses. H5 and H7 influenza viruses are isolated over the world and also circulate among poultry in Asia. In 2010, two H5N1 highly pathogenic avian influenza viruses (HPAIVs) were isolated from fecal samples of water birds on the flyway of migration from Siberia, Russia to the south in Hokkaido, Japan. H7N9 viruses are sporadically isolated from humans and circulate in poultry in China. To monitor whether these viruses have spread in the wild bird population, we conducted virological surveillance of avian influenza in migratory water birds in autumn from 2010 to 2014. A total of 8103 fecal samples from migratory water birds were collected in Japan and Mongolia, and 350 influenza viruses including 13 H5 and 19 H7 influenza viruses were isolated. A phylogenetic analysis revealed that all isolates are genetically closely related to viruses circulating among wild water birds. The results of the antigenic analysis indicated that the antigenicity of viruses in wild water birds is highly stable despite their nucleotide sequence diversity but is distinct from that of HPAIVs recently isolated in Asia. The present results suggest that HPAIVs and Chinese H7N9 viruses were not predominantly circulating in migratory water birds; however, continued monitoring of H5 and H7 influenza viruses both in domestic and wild birds is recommended for the control of avian influenza.

Emergence of H7N9 Influenza A Virus Resistant to Neuraminidase Inhibitors in Nonhuman Primates
Yasushi Itoh, Shintaro Shichinohe, Misako Nakayama, Manabu Igarashi, Akihiro Ishii, Hirohito Ishigaki, Hideaki Ishida, Naoko Kitagawa, Takako Sasamura, Masanori Shiohara, Michiko Doi, Hideaki Tsuchiya, Shinichiro Nakamura, Masatoshi Okamatsu, Yoshihiro Sakoda, Hiroshi Kida, Kazumasa Ogasawara
ANTIMICROBIAL AGENTS AND CHEMOTHERAPY, 59, 8, 4962, 4973, AMER SOC MICROBIOLOGY, 2015年08月, ［査読有り］
英語, 研究論文（学術雑誌）, The number of patients infected with H7N9 influenza virus has been increasing since 2013. We examined the efficacy of neuraminidase (NA) inhibitors and the efficacy of a vaccine against an H7N9 influenza virus, A/Anhui/1/2013 (H7N9), isolated from a patient in a cynomolgus macaque model. NA inhibitors (oseltamivir and peramivir) barely reduced the total virus amount because of the emergence of resistant variants with R289K or 1219T in NA [residues 289 and 219 in N9 of A/Anhui/1/2013 (H7N9) correspond to 292 and 222 in N2, respectively] in three of the six treated macaques, whereas subcutaneous immunization of an inactivated vaccine derived from A/duck/Mongolia/119/2008 (H7N9) prevented propagation of A/Anhui/1/2013 (H7N9) in all vaccinated macaques. The percentage of macaques in which variant H7N9 viruses with low sensitivity to the NA inhibitors were detected was much higher than that of macaques in which variant H5N1 highly pathogenic influenza virus was detected after treatment with one of the NA inhibitors in our previous study. The virus with R289K in NA was reported in samples from human patients, whereas that with 1219T in NA was identified for the first time in this study using macaques, though no variant H7N9 virus was reported in previous studies using mice. Therefore, the macaque model enables prediction of the frequency of emerging H7N9 virus resistant to NA inhibitors in vivo. Since H7N9 strains resistant to NA inhibitors might easily emerge compared to other influenza viruses, monitoring of the emergence of variants is required during treatment of H7N9 influenza virus infection with NA inhibitors.

First Japanese case of muscular dystrophy caused by a mutation in the anoctamin 5 gene
Hiroshi Kida, Ken Sano, Akiko Yorita, Shiroh Miura, Mitsuyoshi Ayabe, Yukiko Hayashi, Ichizo Nishino, Takayuki Taniwaki
NEUROLOGY AND CLINICAL NEUROSCIENCE, 3, 4, 150, 152, WILEY, 2015年07月, ［査読有り］
英語, 研究論文（学術雑誌）, Mutations in the anoctamin 5 gene cause either limb girdle muscular dystrophy or Miyoshi muscular dystrophy 3 in Caucasians. We herein describe a 49-year-old Japanese man with asymmetry of the leg circumference since childhood. Muscle involvement began at the calf muscles, and later spread to the thigh and paraspinal muscles on imaging and physical examination. His external genitalia were morphologically hypoplastic. Genetic analysis identified a novel homozygous mutation, c. 1178G>A (p. Trp393X), in the anoctamin 5 gene. This is the first Japanese case with Miyoshi muscular dystrophy 3 caused by an anoctamin 5 gene mutation (anoctaminopathy). The muscle impairment associated with Miyoshi muscular dystrophy 3 appears to spread from the distal to proximal lower extremities.

豚コレラウイルス非構造蛋白NS4Bの機能と病原性に及ぼす影響　　　　　　　　　　　　　　　
田村友和, Nicolas Ruggli, 長島尚史, 岡松正敏, Artur Summerfield, 松野啓太, 喜田宏, 迫田義博
2015年07月, ［査読有り］
日本語, 研究論文（その他学術会議資料等）

The N-terminal domain of N-pro of classical swine fever virus determines its stability and regulates type I IFN production
Junki Mine, Tomokazu Tamura, Kazuya Mitsuhashi, Masatoshi Okamatsu, Sujira Parchariyanon, Wasana Pinyochon, Nicolas Ruggli, Jon-Duri Tratschin, Hiroshi Kida, Yoshihiro Sakoda
JOURNAL OF GENERAL VIROLOGY, 96, 1746, 1756, SOC GENERAL MICROBIOLOGY, 2015年07月, ［査読有り］
英語, 研究論文（学術雑誌）, The viral protein N-pro is unique to the genus Pestivirus within the family Flaviviridae. After autocatalytic cleavage from the nascent polyprotein, N-pro suppresses type I IFN (IFN-alpha/beta) induction by mediating proteasomal degradation of IFN regulatory factor 3 (IRF-3). Previous studies found that the N-pro-mediated IRF-3 degradation was dependent of a TRASH domain in the C-terminal half of N-pro coordinating zinc by means of the amino acid residues 0112, 0134, D136 and C138. Interestingly, four classical swine fever virus (CSFV) isolates obtained from diseased pigs in Thailand in 1993 and 1998 did not suppress IFN-alpha/beta induction despite the presence of an intact TRASH domain. Through systematic analyses, it was found that an amino acid mutation at position 40 or mutations at positions 17 and 61 in the N-terminal half of N-pro of these four isolates were related to the lack of IRF-3-degrading activity. restoring a histidine at position 40 or both a proline at position 17 and a lysine at position 61 based on the sequence of a functional N-pro contributed to higher stability of the reconstructed N-pro compared with the N-pro from the Thai isolate. This led to enhanced interaction of N-pro with IRF-3 along with its degradation by the proteasome. The results of the present study revealed that amino acid residues in the N-terminal domain of N-pro are involved in the stability of N-pro, in interaction of N-pro with IRF-3 and subsequent degradation of IRF-3, leading to downregulation of IFN-alpha/beta production.

VIRAL GENETIC DETERMINANTS OF CLASSICAL SWINE FEVER VIRUS VIRULENCE　　　　　　　　　　　　　　　
Tomokazu Tamura, Nicolas Ruggli, Naofumi Nagashima, Masatoshi Okamatsu, Martin A. Hofmann, Hiroshi Kida, Autur Summerfield, Yoshihiro Sakoda
2015年06月, ［査読有り］
英語, 研究論文（国際会議プロシーディングス）

Molecular epidemiology of pathogenic Leptospira spp. in the straw-colored fruit bat (Eidolon helvum) migrating to Zambia from the Democratic Republic of Congo
Hirohito Ogawa, Nobuo Koizumi, Aiko Ohnuma, Alisheke Mutemwa, Bernard M. Hang'ombe, Aaron S. Mweene, Ayato Takada, Chihiro Sugimoto, Yasuhiko Suzuki, Hiroshi Kida, Hirofumi Sawa
INFECTION GENETICS AND EVOLUTION, 32, 143, 147, ELSEVIER SCIENCE BV, 2015年06月, ［査読有り］
英語, 研究論文（学術雑誌）, The role played by bats as a potential source of transmission of Leptospira spp. to humans is poorly understood, despite various pathogenic Leptospira spp. being identified in these mammals. Here, we investigated the prevalence and diversity of pathogenic Leptospira spp. that infect the straw-colored fruit bat (Eidolon helvum). We captured this bat species, which is widely distributed in Africa, in Zambia during 2008-2013. We detected the flagellin B gene (flaB) from pathogenic Leptospira spp. in kidney samples from 79 of 529 E. helvum (14.9%) bats. Phylogenetic analysis of 70 flaB fragments amplified from E. helvum samples and previously reported sequences, revealed that 12 of the fragments grouped with Leptospira borgpetersenii and Leptospira kirschneri; however, the remaining 58 flaB fragments appeared not to be associated with any reported species. Additionally, the 16S ribosomal RNA gene (rrs) amplified from 27 randomly chosen flaB-positive samples was compared with previously reported sequences, including bat-derived Leptospira spp. All 27 rrs fragments clustered into a pathogenic group. Eight fragments were located in unique branches, the other 19 fragments were closely related to Leptospira spp. detected in bats. These results show that rrs sequences in bats are genetically related to each other without regional variation, suggesting that Leptospira are evolutionarily well-adapted to bats and have uniquely evolved in the bat population. Our study indicates that pathogenic Leptospira spp. in E. helvum in Zambia have unique genotypes. (C) 2015 The Authors. Published by Elsevier B.V.

Analysis of a pair of END+ and END- viruses derived from the same bovine viral diarrhea virus stock reveals the amino acid determinants in N-pro responsible for inhibition of type I interferon production
Takashi Kozasa, Yuri Abe, Kazuya Mitsuhashi, Tomokazu Tamura, Hiroshi Aoki, Masatoshi Ishimaru, Shigeyuki Nakamura, Masatoshi Okamatsu, Hiroshi Kida, Yoshihiro Sakoda
JOURNAL OF VETERINARY MEDICAL SCIENCE, 77, 5, 511, 518, JAPAN SOC VET SCI, 2015年05月, ［査読有り］
英語, 研究論文（学術雑誌）, The Exaltation of Newcastle disease virus (END) phenomenon is induced by the inhibition of type I interferon in pestivirus-infected cells in vitro, via proteasomal degradation of cellular interferon regulatory factor (IRF)-3 with the property of the viral autoprotease protein N-pro. Reportedly, the amino acid residues in the zinc-binding TRASH motif of N-pro determine the difference in characteristics between END-phenomenon-positive (END) and END-phenomenon-negative (END-) classical swine fever viruses (CSFVs). However, the basic mechanism underlying this function in bovine viral diarrhea virus (BVDV) has not been elucidated from the genomic differences between END+ and END- viruses using reverse genetics till date. In the present study, comparison of complete genome sequences of a pair of END+ and END- viruses isolated from the same virus stock revealed that there were only four amino acid substitutions (D136G, I2623V, D3148G and D3502Y) between two viruses. Based on these differences, viruses with and without mutations at these positions were generated using reverse genetics. The END assay, measurements of induced type I interferon and IRF-3 detection in cells infected with these viruses revealed that the aspartic acid at position 136 in the zinc-binding TRASH motif of N-pro was required to inhibit the production of type I interferon via the degradation of cellular IRF-3, consistently with CSFV.

Fluorescent Immunochromatography for Rapid and Sensitive Typing of Seasonal Influenza Viruses
Akira Sakurai, Katsuyoshi Takayama, Namiko Nomura, Naoki Kajiwara, Masatoshi Okamatsu, Naoki Yamamoto, Tsuruki Tamura, Jitsuho Yamada, Masako Hashimoto, Yoshihiro Sakoda, Yoshihiko Suda, Yukuharu Kobayashi, Hiroshi Kida, Futoshi Shibasaki
PLOS ONE, 10, 2, e0116715, PUBLIC LIBRARY SCIENCE, 2015年02月, ［査読有り］
英語, 研究論文（学術雑誌）, Lateral flow tests also known as Immunochromatography (IC) is an antigen-detection method conducted on a nitrocellulose membrane that can be completed in less than 20 min. IC has been used as an important rapid test for clinical diagnosis and surveillance of influenza viruses, but the IC sensitivity is relatively low (approximately 60%) and the limit of detection (LOD) is as low as 10(3) pfu per reaction. Recently, we reported an improved IC assay using antibodies conjugated with fluorescent beads (fluorescent immunochromatography; FLIC) for subtyping H5 influenza viruses (FLIC-H5). Although the FLIC strip must be scanned using a fluorescent reader, the sensitivity (LOD) is significantly improved over that of conventional IC methods. In addition, the antibodies which are specific against the subtypes of influenza viruses cannot be available for the detection of other subtypes when the major antigenicity will be changed. In this study, we established the use of FLIC to type seasonal influenza A and B viruses (FLIC-AB). This method has improved sensitivity to 100-fold higher than that of conventional IC methods when we used several strains of influenza viruses. In addition, FLIC-AB demonstrated the ability to detect influenza type A and influenza type B viruses from clinical samples with high sensitivity and specificity (Type A: sensitivity 98.7% (74/75), specificity 100% (54/54), Type B: sensitivity 100% (90/90), specificity 98.2% (54/55) in nasal swab samples) in comparison to the results of qRT-PCR. And furthermore, FLIC-AB performs better in the detection of early stage infection (under 13h) than other conventional IC methods. Our results provide new strategies to prevent the early-stage transmission of influenza viruses in humans during both seasonal outbreaks and pandemics.

Mucosal Immunization and Adjuvants
Hideki Hasegawa, Elly van Reit, Hiroshi Kida
INFLUENZA PATHOGENESIS AND CONTROL - VOL II, 386, 371, 380, SPRINGER-VERLAG BERLIN, 2015年, ［査読有り］
英語, 論文集(書籍)内論文, The goal of the influenza vaccine is to prevent influenza virus infection and control the yearly seasonal epidemic and pandemic. However, the presently available parenteral influenza vaccine induces only systemic humoral immunity, which does not prevent influenza virus infection on the mucosal surface. Secretary IgA antibodies play an important role in preventing natural infection. Moreover, the IgA antibody response mediates cross-protection against variant viruses in animal models. Thus, a mucosal influenza vaccine that induces mucosal immunity would be a powerful tool to protect individuals from the influenza virus. Although the function of the mucosal immune system, especially in the respiratory tract, is not completely understood, there are several studies underway to develop mucosal influenza vaccines. Here, we will review current knowledge concerning the induction of IgA, the role of B-cell production of influenza virus specific IgA antibodies in anti-influenza immunity, and the role of humoral memory responses induced upon vaccination.

Multi-colored immunochromatography using nanobeads for rapid and sensitive typing of seasonal influenza viruses
Akira Sakurai, Katsuyoshi Takayama, Namiko Nomura, Naoki Yamamoto, Yoshihiro Sakoda, Yukuharu Kobayashi, Hiroshi Kida, Futoshi Shibasaki
JOURNAL OF VIROLOGICAL METHODS, 209, 62, 68, ELSEVIER SCIENCE BV, 2014年12月, ［査読有り］
英語, 研究論文（学術雑誌）, Immunochromatography (IC) is an antigen-detection assay that plays an important role in the rapid diagnosis of influenza viruses because of its rapid turnaround and ease of use. Despite the usefulness of IC, the limit of detection of common IC kits is as high as 10(3)-10(4) plaque forming units (pfu) per reaction, resulting in their limited sensitivities. Early diagnosis within 24 h would provide more appropriate timing of treatment.
In this study, a multi-colored NanoAct(TM) bead IC was established to detect seasonal influenza viruses. This method has approximately 10-fold higher sensitivity than that of colloidal gold or colored latex bead IC assays, and does not require specific instruments. More notably, NanoAct(Tm) bead IC can distinguish influenza A and B viruses from clinical samples with a straightforward readout composed of colored lines. Our results will provide new strategies for the diagnosis, treatment, and a chance to survey of influenza viruses in developing countries and in the field research. (C) 2014 Elsevier B.V. All rights reserved.

Neuraminidase gene homology contributes to the protective activity of influenza vaccines prepared from the influenza virus library
Ahmad M. Haredy, Hiroshi Yamada, Yoshihiro Sakoda, Masatoshi Okamatsu, Naoki Yamamoto, Takeshi Omasa, Yasuko Mori, Hiroshi Kida, Shigefumi Okamoto, Yoshinobu Okuno, Koichi Yamanishi
JOURNAL OF GENERAL VIROLOGY, 95, 2365, 2371, SOC GENERAL MICROBIOLOGY, 2014年11月, ［査読有り］
英語, 研究論文（学術雑誌）, Whole-virus (WV) vaccines from influenza A/duck/Hokkaido/77 (H3N2), and its reassortant strains H3N4, H3N5 and H3N7, which have the same haemagglutinin (HA) gene but different neuraminidase (NA) genes, were prepared from our influenza virus library. Mice were intranasally immunized with equivalent doses of each vaccine (1-0.01 mu g per mouse). All of the mice that received the highest dose of each vaccine (1 mu g per mouse) showed equivalent high HA-inhibiting (HI) antibody titres and survived the H3N2 challenge viruses. However, mice that received lower doses of vaccine (0.1 or 0.01 mu g per mouse) containing a heterologous NA had lower survival rates than those given the H3N2-based vaccine. The lungs of mice challenged with H3N2 virus showed a significantly higher virus clearance rate when the vaccine contained the homologous NA (N2) versus a heterologous NA, suggesting that NA contributed to the protection, especially when the HI antibody level was low. These results suggested that, even if vaccines prepared for a possible upcoming pandemic do not induce sufficient HI antibodies, WV vaccines can still be effective through other matched proteins such as NA.

Potency of an inactivated influenza vaccine prepared from A/duck/Hong Kong/960/1980 (H6N2) against a challenge with A/duck/Vietnam/OIE-0033/2012 (H6N2) in mice
Tatsuya Nishi, Yoshihiro Sakoda, Masatoshi Okamatsu, Hiroshi Kida
ARCHIVES OF VIROLOGY, 159, 10, 2567, 2574, SPRINGER WIEN, 2014年10月, ［査読有り］
英語, 研究論文（学術雑誌）, H6 influenza viruses are prevalent in domestic and wild birds in Eurasian countries and have been isolated from pigs and a human. To prepare for an influenza pandemic, we have established an influenza virus library consisting of more than 1,300 influenza virus strains, including 144 combinations of 16 hemagglutinin and 9 neuraminidase subtypes. H6 viruses in the library were classified into Early, Group II, Group III, and W312 sublineages and the North America lineage on the basis of their phylogenetic features. Chicken antisera to A/duck/Hong Kong/960/1980 (H6N2) of the Early sublineage broadly reacted with viruses of different sublineages in a hemagglutinin inhibition test. A whole inactivated virus particle vaccine was prepared from A/duck/Hong Kong/960/1980 (H6N2) which was stocked in the influenza virus library. The potency of this vaccine against A/duck/Vietnam/OIE-0033/2012 (H6N2), which belongs to a different sublineage, was evaluated in mice. The test vaccine was sufficiently potent to induce an immune response that reduced the impact of disease caused by a challenge with A/duck/Vietnam/OIE-0033/2012 (H6N2) in mice. The present results indicate that the whole inactivated virus particle vaccine prepared from a virus strain in the influenza virus library is useful as a vaccine against pandemic influenza.

Humoral Immune Response to Influenza A(H1N1)pdm2009 in Patients with Natural Infection and in Vaccine Recipients in the 2009 Pandemic
Takuji Kumagai, Tetsuo Nakayama, Yoshinobu Okuno, Tetsuo Kase, Naoko Nishimura, Takao Ozaki, Akiko Miyata, Eitaro Suzuki, Teruo Okafuji, Takao Okafuji, Hitoshi Ochiai, Nobuo Nagata, Hiroyuki Tsutsumi, Masatoshi Okamatsu, Yoshihiro Sakoda, Hiroshi Kida, Toshiaki Ihara
VIRAL IMMUNOLOGY, 27, 8, 368, 374, MARY ANN LIEBERT, INC, 2014年10月, ［査読有り］
英語, 研究論文（学術雑誌）, The 2009 pandemic H1N1 mainly affected adolescents and children, and most of the elderly in Japan escaped clinical illness. To clarify the role of humoral immunity in the infection, the time kinetics of hemagglutination inhibition (HI), neutralization (NT), and IgG subclass antibody response directed against influenza A(H1N1)pdm2009 were analyzed in three consecutive specimens obtained from 51 young adults and children (group 1) who contracted pandemic influenza and from 74 pediatric clinic employees (group 2) inoculated with pandemic monovalent vaccine. In group 1 patients, 6 and 30 patients had lower HI and NT antibody in the acute phase respectively. Thereafter, HI and NT antibody titers increased fourfold or more in 50 patients with peak response in the third specimens obtained four weeks after the onset. IgG1 in 45 patients, IgG3 in 18 patients, and IgG4 in 29 patients showed elevated responses. Forty (54%) and 70 (95%) subjects in group 2 had positive HI and NT antibodies in the prevaccination samples, with increased antibody responses in the follow-up peaking in the second specimens. Forty of those vaccinated had increased IgG1 responses peaking in the third specimens, whereas elevated IgG3 was observed in 22 recipients with the highest level in the second samples. IgG4 did not show any increase in subjects in group 2. A few participants showed an IgG2 response in both groups. An immunologically naive population contracted influenza with apparent clinical symptoms. However, already primed subjects through subclinical infection elicited the unique pattern of IgG subclass responses by vaccination, which differed from those of naive populations.

The relationship between in vivo antiviral activity and pharmacokinetic parameters of peramivir in influenza virus infection model in mice
Makoto Kodama, Ryu Yoshida, Takahiro Hasegawa, Masaaki Izawa, Mitsutaka Kitano, Kaoru Baba, Takeshi Noshi, Takahiro Seki, Kenichi Okazaki, Masakatsu Tsuji, Takushi Kanazu, Hiroshi Kamimori, Tomoyuki Homma, Masanori Kobayashi, Yoshihiro Sakoda, Hiroshi Kida, Akihiko Sato, Yoshinori Yamano
ANTIVIRAL RESEARCH, 109, 110, 115, ELSEVIER SCIENCE BV, 2014年09月, ［査読有り］
英語, 研究論文（学術雑誌）, The purpose of this study was to investigate the relationship between pharmacokinetic (PK) parameters of intravenous (IV) peramivir and in vivo antiviral activity pharmacodynamic (PD) outcomes in a mouse model of influenza virus infection. Peramivir was administrated to mice in three dosing schedules; once, twice and four times after infection of A/WS/33 (H1N1). The survival rate at day 14 after virus infection was employed as the antiviral activity outcome for analysis. The relationship between day 14 survival and PK parameters, including area under the concentration-time curve (AUC), maximum concentration (C-max) and time that drug concentration exceeds IC95 (T>(IC95)), was estimated using a logistic regression model, and model fitness was evaluated by calculation of the Akaike information criterion (AIC) index. The AIC indices of AUC, C-max and T->IC95 were about 114, 151 and 124, respectively. The AIC of AUC and T->IC95 were smaller than that of C-max. Therefore, both AUC and T->IC95 were the PK parameters that correlated best with the antiviral activity of peramivir IV against influenza virus infection in mice. (C) 2014 Elsevier B.V. All rights reserved.

Molecular, biological, and antigenic characterization of a Border disease virus isolated from a pig during classical swine fever surveillance in Japan
Makoto Nagai, Hiroshi Aoki, Yoshihiro Sakoda, Takashi Kozasa, Kaho Tominaga-Teshima, Junki Mine, Yuri Abe, Tomokazu Tamura, Tsubasa Kobayashi, Kaoru Nishine, Kentaro Tateishi, Yudai Suzuki, Mai Fukuhara, Keitaro Ohmori, Reiko Todaka, Kazuhiko Katayama, Tetsuya Mizutani, Shigeyuki Nakamura, Hiroshi Kida, Junsuke Shirai
JOURNAL OF VETERINARY DIAGNOSTIC INVESTIGATION, 26, 4, 547, 552, SAGE PUBLICATIONS INC, 2014年07月, ［査読有り］
英語, 研究論文（学術雑誌）, In the current study, molecular, biological, and antigenic analyses were performed to characterize Border disease virus (BDV) strain FNK2012-1 isolated from a pig in 2012 in Japan. The complete genome comprises 12,327 nucleotides (nt), including a large open reading frame of 11,685 nt. Phylogenetic analysis revealed that FNK2012-1 was clustered into BDV genotype 1 with ovine strains. FNK2012-1 grew in porcine, bovine, and ovine primary cells and cell lines, but grew better in bovine and ovine cells than in porcine cells. Specific pathogen-free pigs inoculated with FNK2012-1 did not show any clinical signs. Noninoculated contact control pigs also did not show clinical signs and did not seroconvert. The results suggest that FNK2012-1 may be of ruminant origin and is poorly adapted to pigs. Such observations can provide important insights into evidence for infection and transmission of BDV, which may be of ruminant origin, among pigs.

Protective Efficacy of Passive Immunization with Monoclonal Antibodies in Animal Models of H5N1 Highly Pathogenic Avian Influenza Virus Infection
Yasushi Itoh, Reiko Yoshida, Shintaro Shichinohe, Megumi Higuchi, Hirohito Ishigaki, Misako Nakayama, Van Loi Pham, Hideaki Ishida, Mitsutaka Kitano, Masahiko Arikata, Naoko Kitagawa, Yachiyo Mitsuishi, Kazumasa Ogasawara, Hideaki Tsuchiya, Takahiro Hiono, Masatoshi Okamatsu, Yoshihiro Sakoda, Hiroshi Kida, Mutsumi Ito, Le Quynh Mai, Yoshihiro Kawaoka, Hiroko Miyamoto, Mari Ishijima, Manabu Igarashi, Yasuhiko Suzuki, Ayato Takada
PLOS PATHOGENS, 10, 6, e1004192, PUBLIC LIBRARY SCIENCE, 2014年06月, ［査読有り］
英語, 研究論文（学術雑誌）, Highly pathogenic avian influenza (HPAI) viruses of the H5N1 subtype often cause severe pneumonia and multiple organ failure in humans, with reported case fatality rates of more than 60%. To develop a clinical antibody therapy, we generated a human-mouse chimeric monoclonal antibody (MAb) ch61 that showed strong neutralizing activity against H5N1 HPAI viruses isolated from humans and evaluated its protective potential in mouse and nonhuman primate models of H5N1 HPAI virus infections. Passive immunization with MAb ch61 one day before or after challenge with a lethal dose of the virus completely protected mice, and partial protection was achieved when mice were treated 3 days after the challenge. In a cynomolgus macaque model, reduced viral loads and partial protection against lethal infection were observed in macaques treated with MAb ch61 intravenously one and three days after challenge. Protective effects were also noted in macaques under immunosuppression. Though mutant viruses escaping from neutralization by MAb ch61 were recovered from macaques treated with this MAb alone, combined treatment with MAb ch61 and peramivir reduced the emergence of escape mutants. Our results indicate that antibody therapy might be beneficial in reducing viral loads and delaying disease progression during H5N1 HPAI virus infection in clinical cases and combined treatment with other antiviral compounds should improve the protective effects of antibody therapy against H5N1 HPAI virus infection.

Potency of an inactivated influenza vaccine prepared from A/duck/Mongolia/119/2008 (H7N9) against the challenge with A/Anhui/1/2013 (H7N9)
Duc-Huy Chu, Yoshihiro Sakoda, Tatsuya Nishi, Takahiro Hiono, Shintaro Shichinohe, Masatoshi Okamatsu, Hiroshi Kida
VACCINE, 32, 28, 3473, 3479, ELSEVIER SCI LTD, 2014年06月, ［査読有り］
英語, 研究論文（学術雑誌）, H7N9 influenza virus infection in humans was reported in China on March 31, 2013. Humans are immunologically naive to the H7N9 subtype, for which the seasonal influenza vaccine is not effective. Thus, the development of an H7N9 influenza virus vaccine is an urgent issue. To prepare for the emergence of an influenza pandemic, we have established a library comprising more than 1300 influenza virus strains with 144 different combinations of 16 HA and 9 NA subtypes. An H7N9 virus strain isolated from a 35-year-old woman, A/Anhui/1/2013 (H7N9), was found to be antigenically similar to H7N9 influenza viruses isolated from migratory ducks. In the present study, the potency of an inactivated whole virus particle vaccine prepared from an H7N9 low pathogenic avian influenza virus, A/duck/Mongolia/119/2008 (H7N9), selected from the library, was assessed by a challenge with A/Anhui/1/2013 (H7N9). The results indicate that the test vaccine was potent enough to induce sufficient immunity to reduce the impact of disease caused by the challenge with A/Anhui/1/2013 (H7N9) in mice. The present results indicate that an inactivated whole virus particle vaccine prepared from an influenza virus strain stored in the library could be useful as a vaccine strain in case of an influenza pandemic. (C) 2014 Published by Elsevier Ltd.

A chicken influenza virus recognizes fucosylated α2,3 sialoglycan receptors on the epithelial cells lining upper respiratory tracts of chickens.
Hiono T, Okamatsu M, Nishihara S, Takase-Yoden S, Sakoda Y, Kida H
Virology, 456, 131, 138, ACADEMIC PRESS INC ELSEVIER SCIENCE, 2014年05月, ［査読有り］
英語, 研究論文（学術雑誌）, Influenza viruses recognize sialoglycans as receptors. Although viruses isolated form chickens preferentially bind to sialic acid alpha 2,3 galactose (SA alpha 2,3Gal) glycans as do those of ducks, chickens were not experimentally infected with viruses isolated from ducks. A chicken influenza virus, A/chicken/Ibaraki/1/ 2005 (H5N2) (Ck/IBR) bound to fucose-branched SAa2,3Gal glycans, whereas the binding towards linear SAa2,3Gal glycans was weak. On the epithelial cells of the upper respiratory tracts of chickens, fucose-branched SA alpha 2,3Gal glycans were detected, but not linear SA alpha 2,3Gal glycans. The growth of Ck/IBR in MDCK-FUT cells, which were genetically prepared to express fucose-branched SA alpha 2,3Gal glycans, was significantly higher than that in the parental MDCK cells. The present results indicate that fucosebranched SA alpha 2,3Gal glycans existing on the epithelial cells lining the upper respiratory tracts of chickens are critical for recognition by Ck/IBR. (c) 2014 Elsevier Inc. All rights reserved.

N-pro of classical swine fever virus contributes to pathogenicity in pigs by preventing type I interferon induction at local replication sites
Tomokazu Tamura, Naofumi Nagashima, Nicolas Ruggli, Artur Summerfield, Hiroshi Kida, Yoshihiro Sakoda
VETERINARY RESEARCH, 45, 47, BIOMED CENTRAL LTD, 2014年04月, ［査読有り］
英語, 研究論文（学術雑誌）, Classical swine fever (CSF) caused by CSF virus (CSFV) is a highly contagious disease of pigs. The viral protein N-pro of CSFV interferes with alpha-and beta-interferon (IFN-alpha/beta) induction by promoting the degradation of interferon regulatory factor 3 (IRF3). During the establishment of the live attenuated CSF vaccine strain GPE(-), N-pro acquired a mutation that abolished its capacity to bind and degrade IRF3, rendering it unable to prevent IFN-alpha/beta induction. In a previous study, we showed that the GPE(-) vaccine virus became pathogenic after forced serial passages in pigs, which was attributed to the amino acid substitutions T830A in the viral proteins E2 and V2475A and A2563V in NS4B. Interestingly, during the re-adaptation of the GPE(-) vaccine virus in pigs, the IRF3-degrading function of N-pro was not recovered. Therefore, we examined whether restoring the ability of N-pro to block IFN-alpha/beta induction of both the avirulent and moderately virulent GPE(-)-derived virus would enhance pathogenicity in pigs. Viruses carrying the N136D substitution in N-pro regained the ability to degrade IRF3 and suppress IFN-alpha/beta induction in vitro. In pigs, functional N-pro significantly reduced the local IFN-alpha mRNA expression in lymphoid organs while it increased quantities of IFN-alpha/beta in the circulation, and enhanced pathogenicity of the moderately virulent virus. In conclusion, the present study demonstrates that functional N-pro influences the innate immune response at local sites of virus replication in pigs and contributes to pathogenicity of CSFV in synergy with viral replication.

Stimulation with the Aureobasidium pullulans-produced β-glucan effectively induces interferon stimulated genes in macrophage-like cell lines.
Muramatsu D, Kawata K, Aoki S, Uchiyama H, Okabe M, Miyazaki T, Kida H, Iwai A
Scientific reports, 4, 4777, NATURE PUBLISHING GROUP, 2014年04月, ［査読有り］
英語, 研究論文（学術雑誌）, A beta-(1,3),(1,6)-D-glucan produced by A. pullulans (AP-PG) is known to be an immune stimulating agent. In this study, we demonstrate that the stimulation with AP-PG effectively induces the interferon (IFN) stimulated genes (ISGs) in macrophage-like cell lines. The ISGs, Mx1, ISG15, and viperin mRNAs were significantly increased in RAW264.7 cells after stimulation with AP-PG. The stimulation with AP-PG transiently induced IFN-beta mRNA. However, the expression of viperin mRNA was also increased after stimulation with AP-PG even when new protein synthesis was completely blocked by treatment with cycloheximide. Further, in IFN-alpha receptor knockdown RAW264.7 cells, AP-PG stimulation more effectively induced viperin mRNA compared with that of IFN-alpha stimulation. The phosphorylation of Ser 727 in STAT1 involved in the enhancement of STAT1 activation was immediately increased after stimulation with AP-PG. In addition, viperin mRNA expression induced after stimulation with IFN-alpha was significantly increased by combined stimulation with AP-PG. These results suggest that stimulation with AP-PG effectively induces the ISGs through the induction of IFN and the enhancement of STAT1-mediated transcriptional activation.

Heat Shock Protein 70 Modulates Influenza A Virus Polymerase Activity
Rashid Manzoor, Kazumichi Kuroda, Reiko Yoshida, Yoshimi Tsuda, Daisuke Fujikura, Hiroko Miyamoto, Masahiro Kajihara, Hiroshi Kida, Ayato Takada
JOURNAL OF BIOLOGICAL CHEMISTRY, 289, 11, 7599, 7614, AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC, 2014年03月, ［査読有り］
英語, 研究論文（学術雑誌）, Background: It has been shown that heat shock protein 70 (Hsp70) plays a role in influenza A virus replication. Results: A correlation between viral replication/transcription activities and nuclear/cytoplasmic shuttling of Hsp70 was observed. Conclusion: Hsp70 modulates the influenza A virus polymerase activity. Significance: This study, for the first time, suggests that Hsp70 may actually assist in influenza A virus replication.
The role of heat shock protein 70 (Hsp70) in virus replication has been discussed for many viruses. The known suppressive role of Hsp70 in influenza virus replication is based on studies conducted in cells with various Hsp70 expression levels. In this study, we determined the role of Hsp70 in influenza virus replication in HeLa and HEK293T cells, which express Hsp70 constitutively. Co-immunoprecipitation and immunofluorescence studies revealed that Hsp70 interacted with PB2 or PB1 monomers and PB2/PB1 heterodimer but not with the PB1/PA heterodimer or PB2/PB1/PA heterotrimer and translocated into the nucleus with PB2 monomers or PB2/PB1 heterodimers. Knocking down Hsp70 resulted in reduced virus transcription and replication activities. Reporter gene assay, immunofluorescence assay, and Western blot analysis of nuclear and cytoplasmic fractions from infected cells demonstrated that the increase in viral polymerase activity during the heat shock phase was accompanied with an increase in Hsp70 and viral polymerases levels in the nuclei, where influenza virus replication takes place, whereas a reduction in viral polymerase activity was accompanied with an increase in cytoplasmic relocation of Hsp70 along with viral polymerases. Moreover, significantly higher levels of viral genomic RNA (vRNA) were observed during the heat shock phase than during the recovery phase. Overall, for the first time, these findings suggest that Hsp70 may act as a chaperone for influenza virus polymerase, and the modulatory effect of Hsp70 appears to be a sequel of shuttling of Hsp70 between nuclear and cytoplasmic compartments.

Frequent glycan structure mining of influenza virus data revealed a sulfated glycan motif that increased viral infection
Tomomi Ichimiya, Shoko Nishihara, Sayaka Takase-Yoden, Hiroshi Kida, Kiyoko Aoki-Kinoshita
BIOINFORMATICS, 30, 5, 706, 711, OXFORD UNIV PRESS, 2014年03月, ［査読有り］
英語, 研究論文（学術雑誌）, Motivation: It is well known influenza viruses recognize and bind terminal sialic acid (SA) on glycans that are found on the cell surface. In this work, we used a data mining technique to analyze the glycan array data of influenza viruses to find novel glycan structures other than SA that may be involved in viral infection.
Results: In addition to SA structures noted previously, we noted the sulfated structures in the mining results. For verification, we overexpressed the sulfotransferase that is involved in synthesizing these structures, and we performed a viral infection experiment to assess changes in infection in these cells. In our results, we found that there is a 70-fold increase in these cells compared with the control. Thus, we have found a novel pattern in glycan structures that may be involved in viral infection.

Histopathological Evaluation of the Diversity of Cells Susceptible to H5N1 Virulent Avian Influenza Virus
Haru Ogiwara, Fumihiko Yasui, Keisuke Munekata, Asako Takagi-Kamiya, Tsubasa Munakata, Namiko Nomura, Futoshi Shibasaki, Kazuhiko Kuwahara, Nobuo Sakaguchi, Yoshihiro Sakoda, Hiroshi Kida, Michinori Kohara
AMERICAN JOURNAL OF PATHOLOGY, 184, 1, 171, 183, ELSEVIER SCIENCE INC, 2014年01月, ［査読有り］
英語, 研究論文（学術雑誌）, Patients infected with highly pathogenic avian influenza A H5N1 viruses (H5N1 HPAIV) show diffuse alveolar damage. However, the temporal progression of tissue damage and repair after viral infection remains poorly defined. Therefore, we assessed the sequential histopathological characteristics of mouse lung after intranasal infection with H5N1 HPAIV or H1N1 2009 pandemic influenza virus (H1N1 pdm). We determined the amount and localization of virus in the Lung through IHC staining and in situ hybridization. IHC used antibodies raised against the virus protein and antibodies specific for macrophages, type II pneumocytes, or proliferating cell nuclear antigen. In situ hybridization used RNA probes against both viral RNA and mRNA encoding the nucleoprotein and the hemagglutinin protein. H5N1 HPAIV infection and replication were observed in multiple lung cell types and might result in rapid progression of lung injury. Both type II pneumocytes and macrophages proliferated after H5N1 HPAIV infection. However, the abundant macrophages failed to block the viral attack, and proliferation of type II pneumocytes failed to restore the damaged alveoli. In contrast, mice infected with H1N1 pdm exhibited modest proliferation of type II pneumocytes and macrophages and slight alveolar damage. These results suggest that the virulence of H5N1 HPAIV results from the wide range of cell tropism of the virus, excessive virus replication, and rapid development of diffuse alveolar damage.

Genetic Analysis of an H5N2 Highly Pathogenic Avian Influenza Virus Isolated from a Chicken in a Live Bird Market in Northern Vietnam in 2012
Tatsuya Nishi, Masatoshi Okamatsu, Kenji Sakurai, Huy Due Chu, Long Pham Thanh, Long van Nguyen, Nam van Hoang, Diep Nguyen Thi, Yoshihiro Sakoda, Hiroshi Kida
JOURNAL OF VETERINARY MEDICAL SCIENCE, 76, 1, 85, 87, JAPAN SOC VET SCI, 2014年01月, ［査読有り］
英語, 研究論文（学術雑誌）, In August 2012, A/chicken/Vietnam/OIE-2215/2012 (H5N2) was isolated from a chicken in a live bird market (LBM) in Northern Vietnam. Intravenous pathogenicity test revealed that this virus is highly pathogenic in chickens. The PA, HA, NP and M, PB2 and NA, and PB1 and NS genes of the isolate were phylogenetically closely related to those of A/duck/Vietnam/OIE-2202/2012 (H5N1) of Glade 2.3.2.1, A/chicken/Vietnam/OIE-1611/2012 (H9N2) and A/chicken/Vietnam/OIE-2468/2012 (H9N2), respectively. All of these viruses were isolated from birds in LBMs in the same province. These results indicate that A/chicken/Vietnam/OIE-2215/2012 (H5N2) is a genetic reassortant and that surveillance of avian influenza in LBMs and stamping out policy are essential for the eradication of highly pathogenic avian influenza viruses from Asia.

Experimental Infection of Macaques with a Wild Water Bird-Derived Highly Pathogenic Avian Influenza Virus (H5N1)
Tomoko Fujiyuki, Misako Yoneda, Fumihiko Yasui, Takeshi Kuraishi, Shosaku Hattori, Hyun-jeong Kwon, Keisuke Munekata, Yuri Kiso, Hiroshi Kida, Michinori Kohara, Chieko Kai
PLOS ONE, 8, 12, e83551, PUBLIC LIBRARY SCIENCE, 2013年12月, ［査読有り］
英語, 研究論文（学術雑誌）, Highly pathogenic avian influenza virus (HPAIV) continues to threaten human health. Non-human primate infection models of human influenza are desired. To establish an animal infection model with more natural transmission and to determine the pathogenicity of HPAIV isolated from a wild water bird in primates, we administered a Japanese isolate of HPAIV (A/whooper swan/Hokkaido/1/2008, H5N1 clade 2.3.2.1) to rhesus and cynomolgus monkeys, in droplet form, via the intratracheal route. Infection of the lower and upper respiratory tracts and viral shedding were observed in both macaques. Inoculation of rhesus monkeys with higher doses of the isolate resulted in stronger clinical symptoms of influenza. Our results demonstrate that HPAIV isolated from a water bird in Japan is pathogenic in monkeys by experimental inoculation, and provide a new method for HPAIV infection of non-human primate hosts, a good animal model for investigation of HPAIV pathogenicity.

Protection against H5N1 Highly Pathogenic Avian and Pandemic (H1N1) 2009 Influenza Virus Infection in Cynomolgus Monkeys by an Inactivated H5N1 Whole Particle Vaccine
Misako Nakayama, Shintaro Shichinohe, Yasushi Itoh, Hirohito Ishigaki, Mitsutaka Kitano, Masahiko Arikata, Van Loi Pham, Hideaki Ishida, Naoko Kitagawa, Masatoshi Okamatsu, Yoshihiro Sakoda, Takaya Ichikawa, Hideaki Tsuchiya, Shinichiro Nakamura, Quynh Mai Le, Mutsumi Ito, Yoshihiro Kawaoka, Hiroshi Kida, Kazumasa Ogasawara
PLOS ONE, 8, 12, e82740, PUBLIC LIBRARY SCIENCE, 2013年12月, ［査読有り］
英語, 研究論文（学術雑誌）, H5N1 highly pathogenic avian influenza virus (HPAIV) infection has been reported in poultry and humans with expanding clade designations. Therefore, a vaccine that induces immunity against a broad spectrum of H5N1 viruses is preferable for pandemic preparedness. We established a second H5N1 vaccine candidate, A/duck/Hokkaido/Vac-3/2007 (Vac-3), in our virus library and examined the efficacy of inactivated whole particles of this strain against two clades of H5N1 HPAIV strains that caused severe morbidity in cynomolgus macaques. Virus propagation in vaccinated macaques infected with either of the H5N1 HPAIV strains was prevented compared with that in unvaccinated macaques. This vaccine also prevented propagation of a pandemic (H1N1) 2009 virus in macaques. In the vaccinated macaques, neutralization activity, which was mainly shown by anti-hemagglutinin antibody, against H5N1 HPAIVs in plasma was detected, but that against H1N1 virus was not detected. However, neuraminidase inhibition activity in plasma and T-lymphocyte responses in lymph nodes against H1N1 virus were detected. Therefore, cross-clade and heterosubtypic protective immunity in macaques consisted of humoral and cellular immunity induced by vaccination with Vac-3.

Polymerase complex with lysine at position 627 of the PB2 of influenza virus A/Hong Kong/483/97 (H5N1) efficiently transcribes and replicates virus genes in mouse cells
Naoki Yamamoto, Yoshihiro Sakoda, Masatoshi Okamatsu, Hiroshi Kida
VIRUS RESEARCH, 178, 2, 404, 410, ELSEVIER SCIENCE BV, 2013年12月, ［査読有り］
英語, 研究論文（学術雑誌）, Influenza virus A/Hong Kong/483/97 (H5N1) (HK483-K) has the PB2 with lysine at position 627 (PB2-627K) and is highly pathogenic in chickens and mice. On the other hand, the pathogenicity of mutant virus (HK483-E), which was generated by substituting lysine with glutamic acid at the position of the PB2, is lower than that of HK483-K in mice, but is highly pathogenic in chickens. The PB2 is one of the components of heterotrimeric polymerase complex, which plays roles in the transcription and replication of virus genes. Cell-free polymerase assay revealed that intrinsic transcription activity of the polymerase complex with PB2-627K is higher than that of glutamic acid (PB2-627E). In chicken cells, transcription efficiency of the polymerase complex with PB2-627E was not lower than those with PB2-627K, indicating that transcription of virus genes is modulated by some host factors in chicken cells, resulting in high growth. Polymerase complex with PB2-627K efficiently transcribes and replicates virus polymerase genes in mouse cells, leading to high growth of HK483-K compared with that of HK483-E. The results of our experiments clearly suggest that efficient transcription and replication of virus genes by polymerase complex result in the higher pathogenicity in mice. (C) 2013 Elsevier B.V. All rights reserved.

Factors responsible for pathogenicity in chickens of a low-pathogenic H7N7 avian influenza virus isolated from a feral duck
Junki Maruyama, Masatoshi Okamatsu, Kosuke Soda, Yoshihiro Sakoda, Hiroshi Kida
ARCHIVES OF VIROLOGY, 158, 12, 2473, 2478, SPRINGER WIEN, 2013年12月, ［査読有り］
英語, 研究論文（学術雑誌）, Highly pathogenic avian influenza viruses have poly-basic amino acid sequences at the cleavage site in their hemagglutinin (HA). Although this poly-basic region is a prerequisite factor for pathogenicity in chickens, not much is known about additional factors responsible for the acquisition of pathogenicity of the duck influenza virus in chickens. Here, we introduced multiple basic amino acid residues into the HA cleavage site of the A/duck/Hokkaido/Vac-2/2004 (H7N7) strain of avian influenza virus, which has low pathogenicity in chickens; the resultant Vac2sub-P0 strain was not intravenously pathogenic in chickens. In contrast, the Vac2sub-P3 strain, which was recovered from three consecutive passages of Vac2sub-P0 in chicks, was intravenously pathogenic in chickens. Six amino acid substitutions were identified by comparison of the Vac2sub-P3 and Vac2sub-P0 genomic sequences: Lys123Glu in PB2, Asn16Asp in PB1, Glu227Gly and Ile388Thr in HA, Gly228Arg in M1, and Leu46Pro in M2. The results of intravenous inoculations of chickens with recombinant virus indicated that all six amino acid substitutions were required to varying degrees for Vac2sub-P3 pathogenicity, with Glu227Gly and Ile388Thr in HA being particularly essential. These results reveal the roles of additional viral factors in the acquisition of pathogenicity in addition to the previously characterized role of the poly-basic amino acid sequences at the HA cleavage site.

Broad-spectrum detection of H5 subtype influenza a viruses with a new fluorescent immunochromatography system
Akira Sakurai, Katsuyoshi Takayama, Namiko Nomura, Tsubasa Munakata, Naoki Yamamoto, Tsuruki Tamura, Jitsuho Yamada, Masako Hashimoto, Kazuhiko Kuwahara, Yoshihiro Sakoda, Yoshihiko Suda, Yukuharu Kobayashi, Nobuo Sakaguchi, Hiroshi Kida, Michinori Kohara, Futoshi Shibasaki
PLoS ONE, 8, 11, e76753, 11, 2013年11月06日, ［査読有り］
英語, 研究論文（学術雑誌）, Immunochromatography (IC) is an antigen-detection assay that plays an important role in the rapid diagnosis of influenza virus because the protocol is short time and easy to use. Despite the usability of IC, the sensitivity is approximately 103 pfu per reaction. In addition, antigen-antibody interaction-based method cannot be used for the detection of influenza viruses with major antigenic change. In this study, we established the use of fluorescent immunochromatography (FLIC) to detect a broad spectrum of H5 subtype influenza A viruses. This method has improved sensitivity 10-100 fold higher than traditional IC because of the use of fluorescent conjugated beads. Our Type-E FLIC kit detected all of the H5 subtype influenza viruses that were examined, as well as recombinant hemagglutinin (HA) proteins (rHAs) belonging to the Eurasian H5 subtype viruses and the Type-N diagnosed North American H5 subtype influenza A viruses. Thus, this kit has the improved potential to detect H5 subtype influenza viruses of different clades with both Type-E and Type-N FLIC kits. Compared with PCR-based diagnosis, FLIC has a strong advantage in usability, because the sample preparation required for FLIC is only mix-and-drop without any additional steps such as RNA extraction. Our results can provide new strategies against the spread and transmission of HPAI H5N1 viruses in birds and mammals including humans. Copyright: © 2013 Sakurai et al.

Resistance to influenza A virus infection in transformed cell lines expressing an anti-PB2 monoclonal antibody
Yoshikazu Fujimoto, Kinuyo Ozaki, Masahiro Maeda, Ken-ichi Nishijima, Hiroki Takakuwa, Koichi Otsuki, Hiroshi Kida, Etsuro Ono
VETERINARY JOURNAL, 198, 2, 487, 493, ELSEVIER SCI LTD, 2013年11月, ［査読有り］
英語, 研究論文（学術雑誌）, The polymerase basic 2 (PB2) protein is one of four proteins that make up the influenza A virus replication complex, which is responsible for viral gene transcription and replication. To assess the antiviral potential of an anti-PB2 monoclonal antibody that inhibits RNA transcription of influenza A viruses, Mardin-Darby canine kidney (MDCK) cells were transformed with two transgenes that encode the light and heavy chains of the monoclonal antibody. The transformed cell lines expressing this monoclonal antibody displayed resistance to several subtypes of influenza A virus infection. In the transformed cell lines infected with influenza A virus, the level of viral RNA transcription was decreased and the effective nuclear transportation of the PB2 protein was also inhibited. These results demonstrate that the anti-PB2 intrabody is potentially able to interfere with the effective nuclear transportation of PB2 protein, resulting in the observed resistance to influenza A virus infection in vitro. (C) 2013 Elsevier Ltd. All rights reserved.

The genetic and antigenic diversity of avian influenza viruses isolated from domestic ducks, muscovy ducks, and chickens in northern and southern Vietnam, 2010-2012
Masatoshi Okamatsu, Tatsuya Nishi, Naoki Nomura, Naoki Yamamoto, Yoshihiro Sakoda, Kenji Sakurai, Huy Duc Chu, Long Pham Thanh, Long Van Nguyen, Nam Van Hoang, Tien Ngoc Tien, Reiko Yoshida, Ayato Takada, Hiroshi Kida
VIRUS GENES, 47, 2, 317, 329, SPRINGER, 2013年10月, ［査読有り］
英語, 研究論文（学術雑誌）, To estimate the prevalence of avian influenza virus infection in Vietnam, surveillance was conducted in domestic and wild birds from households, live-bird markets, slaughtering sites, and bird sanctuaries in Vietnam between October 2010 and October 2012. Of the 4,550 samples collected, 226 influenza A virus isolates were obtained from domestic ducks, muscovy ducks, and chickens. Of these, 25 and 22 H5N1 highly pathogenic avian influenza viruses (HPAIVs) were isolated from apparently healthy domestic ducks in live-bird markets and slaughtering sites in northern and southern Vietnam, respectively. The HA genes of H5 viruses isolated from birds in northern Vietnam phylogenetically belonged to the genetic clade 2.3.2.1 and those in southern Vietnam belonged to the genetic clade 1.1. In addition, 39 H3, 12 H4, 1 H5, 93 H6, 2 H7, 18 H9, 3 H10, and 11 H11 viruses were isolated. Phylogenetic and antigenic analyses of the H6 and H9 viruses revealed that they were closely related to the isolates obtained from domestic poultry in China. Phylogenetic analyses of internal gene segments of these isolates revealed that these viruses were circulating in both domestic and wild birds in Asia and reassortment events had occurred frequently. Therefore, it will be important to continue the surveillance and strict controls over the movement and trade of poultry and poultry products in order to eradicate H5N1 HPAIV from Asia.

Pathogenicity of Pandemic H1N1 Influenza A Virus in Immunocompromised Cynomolgus Macaques
Van Loi Pham, Misako Nakayama, Yasushi Itoh, Hirohito Ishigaki, Mitsutaka Kitano, Masahiko Arikata, Hideaki Ishida, Naoko Kitagawa, Shintaro Shichinohe, Masatoshi Okamatsu, Yoshihiro Sakoda, Hideaki Tsuchiya, Shinichiro Nakamura, Hiroshi Kida, Kazumasa Ogasawara
PLOS ONE, 8, 9, e75910, PUBLIC LIBRARY SCIENCE, 2013年09月, ［査読有り］
英語, 研究論文（学術雑誌）, Pandemic (H1N1) 2009 influenza virus spread throughout the world since most people did not have immunity against the virus. In the post pandemic phase when many humans might possess immunity against the pandemic virus, one of the concerns is infection in immunocompromised people. Therefore, we used an immunosuppressed macaque model to examine pathogenicity of the pandemic (H1N1) 2009 virus under an immunocompromised condition. The virus in nasal samples of immunosuppressed macaques infected with the pandemic (H1N1) 2009 virus was detected longer after infection than was the virus in nasal samples of immunocompetent macaques. As expected, not only virus amounts but also virus propagation sites in the immunosuppressed macaques were larger than those in lungs of the immunocompetent macaques when they were infected with the pandemic virus. Immunosuppressed macaques possessed low levels of immune cells producing cytokines and chemokines, but levels of inflammatory cytokines/chemokine interleukin (IL)-6, IL-18, and monocyte chemotactic protein (MCP)-1 in lungs of the immunosuppressed macaques were higher than those in lungs of the immunocompetent macaques, though the differences were not statistically significant. Therefore, under an immunosuppressive condition, the pandemic influenza (H1N1) 2009 virus might cause more severe morbidity with high cytokine/chemokine production by the host innate immune system than that seen in macaques under the immunocompetent condition.

Selection of H3 avian influenza viruses with SAα2,6Gal receptor specificity in pigs.
Shichinohe S, Okamatsu M, Sakoda Y, Kida H
Virology, 444, 1-2, 404, 408, ACADEMIC PRESS INC ELSEVIER SCIENCE, 2013年09月, ［査読有り］
英語, 研究論文（学術雑誌）, Avian influenza viruses possess hemagglutinin (HA) which preferentially bind to the sialic acid alpha 2,3-galactose sialyloligosaccharides (SA alpha 2,3Gal) receptor. In contrast, human influenza viruses bind to sialic acid alpha 2,6-galactose sialyloligosaccharides (SA alpha 2,6Gal). The A/Hong Kong/68 (H3N2) virus preferentially binds to SA alpha 2,6Gal, although its HA gene was derived from an avian influenza virus strain. To elucidate the mechanisms behind acquisition of binding specificity for the human-type receptor, the avian influenza virus, A/duck/Hokkaido/5/77 (H3N2), which carries the HA with SA alpha 2,3Gal receptor specificity, was consecutively passaged in pigs. Viruses that preferentially bind to the SA alpha 2,6Gal receptor were predominantly recovered from the nasal swabs of pigs after three passages. The present results indicate that avian influenza Viruses can acquire the potential to infect humans after multiple infections in a pig population. Intensive surveillance of swine influenza is, thus, important for the preparedness for the future pandemics. (C) 2013 Elsevier Inc. All rights reserved.

Characterization of H7N9 influenza A viruses isolated from humans
Tokiko Watanabe, Maki Kiso, Satoshi Fukuyama, Noriko Nakajima, Masaki Imai, Shinya Yamada, Shin Murakami, Seiya Yamayoshi, Kiyoko Iwatsuki-Horimoto, Yoshihiro Sakoda, Emi Takashita, Ryan McBride, Takeshi Noda, Masato Hatta, Hirotaka Imai, Dongming Zhao, Noriko Kishida, Masayuki Shirakura, Robert P. de Vries, Shintaro Shichinohe, Masatoshi Okamatsu, Tomokazu Tamura, Yuriko Tomita, Naomi Fujimoto, Kazue Goto, Hiroaki Katsura, Eiryo Kawakami, Izumi Ishikawa, Shinji Watanabe, Mutsumi Ito, Yuko Sakai-Tagawa, Yukihiko Sugita, Ryuta Uraki, Reina Yamaji, Amie J. Eisfeld, Gongxun Zhong, Shufang Fan, Jihui Ping, Eileen A. Maher, Anthony Hanson, Yuko Uchida, Takehiko Saito, Makoto Ozawa, Gabriele Neumann, Hiroshi Kida, Takato Odagiri, James C. Paulson, Hideki Hasegawa, Masato Tashiro, Yoshihiro Kawaoka
NATURE, 501, 7468, 551, +, NATURE PUBLISHING GROUP, 2013年09月, ［査読有り］
英語, 研究論文（学術雑誌）, Avian influenza A viruses rarely infect humans; however, when human infection and subsequent human-to-human transmission occurs, worldwide outbreaks (pandemics) can result. The recent sporadic infections of humans in China with a previously unrecognized avian influenza A virus of the H7N9 subtype (A(H7N9)) have caused concern owing to the appreciable case fatality rate associated with these infections (more than 25%), potential instances of human-to-human transmission(1), and the lack of pre-existing immunity among humans to viruses of this subtype. Here we characterize two early human A(H7N9) isolates, A/Anhui/1/2013 (H7N9) and A/Shanghai/1/2013 (H7N9); hereafter referred to as Anhui/1 and Shanghai/1, respectively. In mice, Anhui/1 and Shanghai/1 were more pathogenic than a control avian H7N9 virus (A/duck/Gunma/466/2011 (H7N9); Dk/GM466) and a representative pandemic 2009 H1N1 virus (A/California/4/2009 (H1N1pdm09); CA04). Anhui/1, Shanghai/1 and Dk/GM466 replicated well in the nasal turbinates of ferrets. In nonhuman primates, Anhui/1 and Dk/GM466 replicated efficiently in the upper and lower respiratory tracts, whereas the replicative ability of conventional human influenza viruses is typically restricted to the upper respiratory tract of infected primates. By contrast, Anhui/1 did not replicate well in miniature pigs after intranasal inoculation. Critically, Anhui/1 transmitted through respiratory droplets in one of three pairs of ferrets. Glycan arrays showed that Anhui/1, Shanghai/1 and A/Hangzhou/1/2013 (H7N9) (a third human A(H7N9) virus tested in this assay) bind to human virus-type receptors, a property that may be critical for virus transmissibility in ferrets. Anhui/1 was found to be less sensitive in mice to neuraminidase inhibitors than a pandemic H1N1 2009 virus, although both viruses were equally susceptible to an experimental antiviral polymerase inhibitor. The robust replicative ability in mice, ferrets and nonhuman primates and the limited transmissibility in ferrets of Anhui/1 suggest that A(H7N9) viruses have pandemic potential.

Excessive Cytokine Response to Rapid Proliferation of Highly Pathogenic Avian Influenza Viruses Leads to Fatal Systemic Capillary Leakage in Chickens
Saya Kuribayashi, Yoshihiro Sakoda, Takeshi Kawasaki, Tomohisa Tanaka, Naoki Yamamoto, Masatoshi Okamatsu, Norikazu Isoda, Yoshimi Tsuda, Yuji Sunden, Takashi Umemura, Noriko Nakajima, Hideki Hasegawa, Hiroshi Kida
PLOS ONE, 8, 7, e68375, PUBLIC LIBRARY SCIENCE, 2013年07月, ［査読有り］
英語, 研究論文（学術雑誌）, Highly pathogenic avian influenza viruses (HPAIVs) cause lethal infection in chickens. Severe cases of HPAIV infections have been also reported in mammals, including humans. In both mammals and birds, the relationship between host cytokine response to the infection with HPAIVs and lethal outcome has not been well understood. In the present study, the highly pathogenic avian influenza viruses A/turkey/Italy/4580/1999 (H7N1) (Ty/Italy) and A/chicken/Netherlands/2586/2003 (H7N7) (Ck/NL) and the low pathogenic avian influenza virus (LPAIV) A/chicken/Ibaraki/1/2005 (H5N2) (Ck/Ibaraki) were intranasally inoculated into chickens. Ty/Italy replicated more extensively than Ck/NL in systemic tissues of the chickens, especially in the brain, and induced excessive mRNA expression of inflammatory and antiviral cytokines (IFN-gamma, IL-1 beta, IL-6, and IFN-alpha) in proportion to its proliferation. Using in situ hybridization, IL-6 mRNA was detected mainly in microglial nodules in the brain of the chickens infected with Ty/Italy. Capillary leakage assessed by Evans blue staining was observed in multiple organs, especially in the brains of the chickens infected with Ty/Italy, and was not observed in those infected with Ck/NL. In contrast, LPAIV caused only local infection in the chickens, with neither apparent cytokine expression nor capillary leakage in any tissue of the chickens. The present results indicate that an excessive cytokine response is induced by rapid and extensive proliferation of HPAIV and causes fatal multiple organ failure in chickens.

An MDCK cell culture-derived formalin-inactivated influenza virus whole-virion vaccine from an influenza virus library confers cross-protective immunity by intranasal administration in mice
Ahmad M. Haredy, Nobuyuki Takenaka, Hiroshi Yamada, Yoshihiro Sakoda, Masatoshi Okamatsu, Naoki Yamamoto, Takeshi Omasa, Hisao Ohtake, Yasuko Mori, Hiroshi Kida, Koichi Yamanishi, Shigefumi Okamoto
Clinical and Vaccine Immunology, 20, 7, 998, 1007, 2013年07月, ［査読有り］
英語, 研究論文（学術雑誌）, It is currently impossible to predict the next pandemic influenza virus strain. We have thus established a library of influenza viruses of all hemagglutinin and neuraminidase subtypes and their genes. In this article, we examine the applicability of a rapid production model for the preparation of vaccines against emerging pandemic influenza viruses. This procedure utilizes the influenza virus library, cell culture-based vaccine production, and intranasal administration to induce a cross-protective immune response. First, an influenza virus reassortant from the library, A/duck/Hokkaido/Vac-3/2007 (H5N1), was passaged 22 times (P22) in Madin-Darby canine kidney (MDCK) cells. The P22 virus had a titer of >
2 × 108PFU/ml, which was 40 times that of the original strain, with 4 point mutations, which altered amino acids in the deduced protein sequences encoded by the PB2 and PA genes. We then produced a formalin-inactivated whole-virion vaccine from the MDCK cell-cultured A/duck/Hokkaido/Vac-3/2007 (H5N1) P22 virus. Intranasal immunization of mice with this vaccine protected them against challenges with lethal influenza viruses of homologous and heterologous subtypes. We further demonstrated that intranasal immunization with the vaccine induced cross-reactive neutralizing antibody responses against the homotypic H5N1 influenza virus and its antigenic variants and cross-reactive cell-mediated immune responses to the homologous virus, its variants within a subtype, and even an influenza virus of a different subtype. These results indicate that a rapid model for emergency vaccine production may be effective for producing the next generation of pandemic influenza virus vaccines. Copyright © 2013, American Society for Microbiology. All Rights Reserved.

Proteins of duck influenza virus responsible for acquisition of pathogenicity in chickens
Naoki Yamamoto, Kosuke Soda, Yoshihiro Sakoda, Masatoshi Okamatsu, Hiroshi Kida
VIRUS RESEARCH, 173, 2, 294, 298, ELSEVIER SCIENCE BV, 2013年05月, ［査読有り］
英語, 研究論文（学術雑誌）, Influenza virus rgVac1 sub-P0 (H5N1) (rgVac1-P0), in which a pair of dibasic,amino acid residues was introduced at the cleavage site of the HA of a reassortant of H5N2 and H7N1 viruses of duck origin, was low pathogenic in chickens. Vac1 sub-P3 (H5N1) (Vac1-P3) was selected as a highly pathogenic avian influenza virus by 3 consecutive passages in chickens from low pathogenic strain rgVac1-P0. Comparison of amino acid sequences of the virus proteins and experimental infection of chickens with a series of recombinant viruses demonstrated that in addition to the HA, each of the PA, NP, M1, and M2 of Vac1-P3 are responsible for the acquisition of pathogenicity in chickens. These 4 proteins of Vac1-P3 synergistically contributed to efficient virus replication in chickens. (C) 2013 Elsevier B.V. All rights reserved.

Potency of an inactivated influenza vaccine prepared from a non-pathogenic H5N1 virus against a challenge with antigenically drifted highly pathogenic avian influenza viruses in chickens
Shintaro Shichinohe, Masatoshi Okamatsu, Naoki Yamamoto, Yu Noda, Yuka Nomoto, Takashi Honda, Noriyasu Takikawa, Yoshihiro Sakoda, Hiroshi Kida
VETERINARY MICROBIOLOGY, 164, 1-2, 39, 45, ELSEVIER SCIENCE BV, 2013年05月, ［査読有り］
英語, 研究論文（学術雑誌）, Antigenic variants of H5N1 highly pathogenic avian influenza virus (HPAIV) have selected and are prevailing in poultry populations in Asia. In the present study, the potency of inactivated influenza vaccine prepared from a non-pathogenic H5N1 avian influenza virus, A/duck/Hokkaido/Vac-3/2007 (H5N1), was assessed by challenging with H5N1 HPAIV variants, A/muscovy duck/Vietnam/OIE-559/2011 (H5N1), A/whooper swan/Hokkaido/4/2011 (H5N1), and A/peregrine falcon/Hong Kong/810/2009 (H5N1) belonging to clades 1, 23.2.1, and 2.3.4, respectively. All chickens immunized with the Vac-3 vaccine survived without showing any clinical signs after intranasal challenge either with A/whooper swan/Hokkaido/4/2011 (H5N1) or A/muscovy duck/Vietnam/OIE-559/2011 (H5N1). After challenge with A/peregrine falcon/Hong Kong/810/2009 (H5N1), 10 out of 12 vaccinated chickens survived and the other 2 died on 4 or 7 post-challenge days. The Vac-3 vaccine of 2.4-fold antigen concentration conferred complete protective immunity in chickens against challenge with A/peregrine falcon/Hong Kong/810/2009 (H5N1). (C) 2013 Elsevier B.V. All rights reserved.

Inhibition of influenza A virus infection by Galectin-9　　　　　　　　　　　　　　　
T. Hattori, J. Maruyama, Y. Fujioka, Y. Nakayama, Y. Ohba, T. Niki, T. Arikawa, T. Miyazaki, M. Hirashima, H. Kida
Jpn. J. Vet. Res., 61, 1&2, 5, 18, 2013年05月, ［査読有り］
英語, 研究論文（学術雑誌）

Antiviral activity of stachyflin on influenza A viruses of different hemagglutinin subtypes
Yurie Motohashi, Manabu Igarashi, Masatoshi Okamatsu, Takeshi Noshi, Yoshihiro Sakoda, Naoki Yamamoto, Kimihito Ito, Ryu Yoshida, Hiroshi Kida
VIROLOGY JOURNAL, 10, 118, BIOMED CENTRAL LTD, 2013年04月, ［査読有り］
英語, 研究論文（学術雑誌）, Background: The hemagglutinin (HA) of influenza viruses is a possible target for antiviral drugs because of its key roles in the initiation of infection. Although it was found that a natural compound, Stachyflin, inhibited the growth of H1 and H2 but not H3 influenza viruses in MDCK cells, inhibitory activity of the compound has not been assessed against H4-H16 influenza viruses and the precise mechanism of inhibition has not been clarified.
Methods: Inhibitory activity of Stachyflin against H4-H16 influenza viruses, as well as H1-H3 viruses was examined in MDCK cells. To identify factors responsible for the susceptibility of the viruses to this compound, Stachyflin-resistant viruses were selected in MDCK cells and used for computer docking simulation.
Results: It was found that in addition to antiviral activity of Stachyflin against influenza viruses of H1 and H2 subtypes, it inhibited replication of viruses of H5 and H6 subtypes, as well as A(H1N1)pdm09 virus in MDCK cells. Stachyflin also inhibited the virus growth in the lungs of mice infected with A/WSN/1933 (H1N1) and A/chicken/Ibaraki/1/2005 (H5N2). Substitution of amino acid residues was found on the HA2 subunit of Stachyflin-resistant viruses. Docking simulation indicated that D37, K51, T107, and K121 are responsible for construction of the cavity for the binding of the compound. In addition, 3-dimensional structure of the cavity of the HA of Stachyflin-susceptible virus strains was different from that of insusceptible virus strains.
Conclusion: Antiviral activity of Stachyflin was found against A(H1N1) pdm09, H5, and H6 viruses, and identified a potential binding pocket for Stachyflin on the HA. The present results should provide us with useful information for the development of HA inhibitors with more effective and broader spectrum.

Novel thiosialosides tethered to metal nanoparticles as potent influenza A virus haemagglutinin blockers.
Feng F, Sakoda Y, Ohyanagi T, Nagahori N, Shibuya H, Okamastu M, Miura N, Kida H, Nishimura SI
Antiviral therapy, 2013年02月, ［査読有り］

The PB2, PA, HA, NP, and NS genes of a highly pathogenic avian influenza virus A/whooper swan/Mongolia/3/2005 (H5N1) are responsible for pathogenicity in ducks
Masahiro Kajihara, Yoshihiro Sakoda, Kosuke Soda, Kenji Minari, Masatoshi Okamatsu, Ayato Takada, Hiroshi Kida
VIROLOGY JOURNAL, 10, 45, BIOMED CENTRAL LTD, 2013年02月, ［査読有り］
英語, 研究論文（学術雑誌）, Background: Wild ducks are the natural hosts of influenza A viruses. Duck influenza, therefore, has been believed inapparent infection with influenza A viruses, including highly pathogenic avian influenza viruses (HPAIVs) in chickens. In fact, ducks experimentally infected with an HPAIV strain, A/Hong Kong/483/1997 (H5N1) (HK483), did not show any clinical signs. Another HPAIV strain, A/whooper swan/Mongolia/3/2005 (H5N1) (MON3) isolated from a dead swan, however, caused neurological dysfunction and death in ducks.
Method: To understand the mechanism whereby MON3 shows high pathogenicity in ducks, HK483, MON3, and twenty-four reassortants generated between these two H5N1 viruses were compared for their pathogenicity in domestic ducks.
Results: None of the ducks infected with MON3-based single-gene reassortants bearing the PB2, NP, or NS gene segment of HK483 died, and HK483-based single-gene reassortants bearing PB2, NP, or NS genes of MON3 were not pathogenic in ducks, suggesting that multiple gene segments contribute to the pathogenicity of MON3 in ducks. All the ducks infected with the reassortant bearing PB2, PA, HA, NP, and NS gene segments of MON3 died within five days post-inoculation, as did those infected with MON3. Each of the viruses was assessed for replication in ducks three days post-inoculation. MON3 and multi-gene reassortants pathogenic in ducks were recovered from all of the tissues examined and replicated with high titers in the brains and lungs.
Conclusion: The present results indicate that multigenic factors are responsible for efficient replication of MON3 in ducks. In particular, virus growth in the brain might correlate with neurological dysfunction and the disease severity.

Potency of a vaccine prepared from A/swine/Hokkaido/2/1981 (H1N1) against A/Narita/1/2009 (H1N1) pandemic influenza virus strain
Masatoshi Okamatsu, Yoshihiro Sakoda, Takahiro Hiono, Naoki Yamamoto, Hiroshi Kida
VIROLOGY JOURNAL, 10, 47, BIOMED CENTRAL LTD, 2013年02月, ［査読有り］
英語, 研究論文（学術雑誌）, Background: The pandemic 2009 (H1N1) influenza virus has spread throughout the world and is now causing seasonal influenza. To prepare for the emergence of pandemic influenza, we have established a library of virus strains isolated from birds, pigs, and humans in global surveillance studies.
Methods: Inactivated whole virus particle (WV) and ether-split (ES) vaccines were prepared from an influenza virus strain, A/swine/Hokkaido/2/1981 (H1N1), from the library and from A/Narita/1/2009 (H1N1) pandemic strain. Each of the vaccines was injected subcutaneously into mice and their potencies were evaluated by challenge with A/Narita/1/2009 (H1N1) virus strain in mice.
Results: A/swine/Hokkaido/2/81 (H1N1), which was isolated from the lung of a diseased piglet, was selected on the basis of their antigenicity and growth capacity in embryonated chicken eggs. Two injections of the WV vaccine induced an immune response in mice, decreasing the impact of disease caused by the challenge with A/Narita/1/2009 (H1N1), as did the vaccine prepared from the homologous strain.
Conclusion: The WV vaccine prepared from an influenza virus in the library is useful as an emergency vaccine in the early phase of pandemic influenza.

Fluorescence polarization-based assay using N-glycan-conjugated quantum dots for screening in hemagglutinin blockers for influenza A viruses
Masatoshi Okamatsu, Fei Feng, Tatsuya Ohyanagi, Noriko Nagahori, Kazuhiko Someya, Yoshihiro Sakoda, Nobuaki Miura, Shin-Ichiro Nishimura, Hiroshi Kida
JOURNAL OF VIROLOGICAL METHODS, 187, 2, 390, 394, ELSEVIER SCIENCE BV, 2013年02月, ［査読有り］
英語, 研究論文（学術雑誌）, Attachment of influenza virus to susceptible cells is mediated by viral protein hemagglutinin (HA), which recognizes cell surface glycoconjugates that terminate in alpha-sialosides. To develop anti-influenza drugs based on inhibition of HA-mediated infection, novel fluorescent nanoparticles displaying multiple biantennary N-glycan chains with alpha-sialosides (A2-PC-QDs) that have high affinity for the HA were designed and constructed. The A2-PC-QDs enabled an easy and efficient fluorescence polarization (FP) assay for detection of interaction with the HA and competitive inhibition even by small molecule compounds against A2-PC-QDs-HA binding. The quantum dot (QD)-based FP assay established in the present study is a useful tool for high-throughput screening and to accelerate the development of novel and more effective blockers of the viral attachment of influenza virus. (C) 2012 Elsevier B.V. All rights reserved.

Overcoming chemoresistance of small-cell lung cancer through stepwise HER2-targeted antibody-dependent cell-mediated cytotoxicity and VEGF-targeted antiangiogenesis.
Toshiyuki Minami, Takashi Kijima, Satoshi Kohmo, Hisashi Arase, Yasushi Otani, Izumi Nagatomo, Ryo Takahashi, Kotaro Miyake, Masayoshi Higashiguchi, Osamu Morimura, Shoichi Ihara, Kazuyuki Tsujino, Haruhiko Hirata, Koji Inoue, Yoshito Takeda, Hiroshi Kida, Isao Tachibana, Atsushi Kumanogoh
Scientific reports, 3, 2669, 2669, 2013年, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, Small-cell lung cancer (SCLC) easily recurs with a multidrug resistant phenotype. However, standard therapeutic strategies for relapsed SCLC remain unestablished. We found that human epidermal growth factor receptor 2 (HER2) is not only expressed in pretreated human SCLC specimens, but is also upregulated when HER2-positive SCLC cells acquire chemoresistance. Trastuzumab induced differential levels of antibody-dependent cell-mediated cytotoxicity (ADCC) to HER2-positive SCLC cells. Furthermore, as a mechanism of the differential levels of ADCC, we have revealed that coexpression of intracellular adhesion molecule (ICAM)-1 on SCLC cells is essential to facilitate and accelerate the trastuzumab-mediated ADCC. Although SN-38-resistant SCLC cells lacking ICAM-1 expression were still refractory to trastuzumab, their in vivo growth was significantly suppressed by bevacizumab treatment due to dependence on their distinctive and abundant production of vascular endothelial growth factor. Collectively, stepwise treatment with trastuzumab and bevacizumab is promising for the treatment of chemoresistant SCLC.

Novel thiosialosides tethered to metal nanoparticles as potent influenza A virus haemagglutinin blockers
Fei Feng, Yoshihiro Sakoda, Tatsuya Ohyanagi, Noriko Nagahori, Hitomi Shibuya, Masatoshi Okamastu, Nobuaki Miura, Hiroshi Kida, Shin-Ichiro Nishimura
Antiviral Chemistry and Chemotherapy, 23, 2, 59, 65, 2013年, ［査読有り］
英語, 研究論文（学術雑誌）, Background: The purpose of this study was to develop a new class of influenza A virus haemagglutinin (HA) blockers by tethering thiosialoside molecules to metal nanoparticles and producing glycoclusters that enhance the affinity of HA binding by N-acetylneuraminic acid. Methods: Oxygen of the glycoside bond of sialoside was replaced with sulfur to prevent hydrolytic digestion of the N-acetylneuraminic acid residue by viral neuraminidase. Two novel thiosialosides, α-2-S-[p-(N-levulinyl) aminophenyl]-5-N-acetylneuraminic acid (Neu5Ac-S-Lev) and α-2-S-[m-(N-levulinyl)aminobenzyl]-5-Nacetylneuraminic acid (Neu5Ac-S-CH 2-Lev), were tethered onto the surface of metal nanoparticles via an aminooxy functionalized thiol linker in a glycoblotting reaction. Gold (Au) and silver (Ag) nanoparticles were coated simultaneously with 11-mercaptoundecyl phosphorylcholine to reduce non-specific adsorption of proteins. Phosphorylcholine self-assembled monolayercoated metals displaying clustered Neu5Ac (Neu5Ac-PCSAM-Au and Neu5Ac-PCSAM-Ag) were subjected to haemagglutination inhibition (HI) assays using the influenza A virus strain A/PR/8/1934 (H1N1). Results: Glyconanoparticles with thiosialosides had potent HI activities. In particular, Neu5Ac-PCSAM-Au with a diameter of 20 nm corresponding to 9.8 μM monosaccharide Neu5Ac was the most potent HA inhibitor. The versatility of this strategy was demonstrated by similar submicromolar HI activities of Neu5Ac-PCSAM--Ag with diameters of 50 nm and 150 nm. Conclusions: Glycosylated metal nanoparticles were designed and synthesized as potent influenza A virus HA blockers. This study may contribute to the acceleration of the discovery of a new class of nanoparticle antiinfluenza drugs. © 2013 International Medical Press.

タイプ１インターフェロンはインフルエンザの病状を決定する肺細胞上のFasLの発現を制御する
Fujikura D, Chiba S, Muramatsu D, Kazumata M, Nakayama Y, Kawai T, Akira S, Kida H, Miyazaki T
PloS one, 8, e55321, 2, 2013年, ［査読有り］

The nucleoprotein is responsible for intracerebral pathogenicity of A/duck/Mongolia/47/2001 (H7N1) in chicks
Norikazu Isoda, Yoshimi Tsuda, Shingo Asakura, Masatoshi Okamatsu, Yoshihiro Sakoda, Hiroshi Kida
ARCHIVES OF VIROLOGY, 157, 12, 2257, 2264, SPRINGER WIEN, 2012年12月, ［査読有り］
英語, 研究論文（学術雑誌）, Avian influenza viruses A/duck/Mongolia/47/2001 (H7N1) (47/01) and A/duck/Mongolia/867/2002 (H7N1) (867/02) were defined as low-pathogenic avian influenza viruses (LPAIVs) using an intravenous pathogenicity test in chickens. On the other hand, the intracerebral pathogenicity indices of 47/01 and 867/02 were 1.30 and 0.00, respectively. A series of reassortant viruses were generated between 47/01 and 867/02, and their intracerebral pathogenicity was compared in one-day-old chicks to identify the protein(s) responsible for the intracerebral pathogenicity of 47/01. The results indicate that the amino acids at positions 50 and 98 of the nucleoprotein are related to the pathogenicity of 47/01 in chicks by intracerebral inoculation. A significant association was found between mortality of the chicks inoculated intracerebrally with 47/01 and virus replication in the lungs and/or brain. These results indicate that the NP of avian influenza viruses may be responsible for intracerebral pathogenicity in the host.

Selection of Classical Swine Fever Virus with Enhanced Pathogenicity Reveals Synergistic Virulence Determinants in E2 and NS4B
Tomokazu Tamura, Yoshihiro Sakoda, Fumi Yoshino, Takushi Nomura, Naoki Yamamoto, Yuka Sato, Masatoshi Okamatsu, Nicolas Ruggli, Hiroshi Kida
JOURNAL OF VIROLOGY, 86, 16, 8602, 8613, AMER SOC MICROBIOLOGY, 2012年08月, ［査読有り］
英語, 研究論文（学術雑誌）, Classical swine fever virus (CSFV) is the causative agent of classical swine fever (CSF), a highly contagious disease of pigs. There are numerous CSFV strains that differ in virulence, resulting in clinical disease with different degrees of severity. Low-virulent and moderately virulent isolates cause a mild and often chronic disease, while highly virulent isolates cause an acute and mostly lethal hemorrhagic fever. The live attenuated vaccine strain GPE(-) was produced by multiple passages of the virulent ALD strain in cells of swine, bovine, and guinea pig origin. With the aim of identifying the determinants responsible for the attenuation, the GPE- vaccine virus was readapted to pigs by serial passages of infected tonsil homogenates until prolonged viremia and typical signs of CSF were observed. The GPE(-)/P-11 virus isolated from the tonsils after the 11th passage in vivo had acquired 3 amino acid substitutions in E2 (T830A) and NS4B (V2475A and A2563V) compared with the virus before passages. Experimental infection of pigs with the mutants reconstructed by reverse genetics confirmed that these amino acid substitutions were responsible for the acquisition of pathogenicity. Studies in vitro indicated that the substitution in E2 influenced virus spreading and that the changes in NS4B enhanced the viral RNA replication. In conclusion, the present study identified residues in E2 and NS4B of CSFV that can act synergistically to influence virus replication efficiency in vitro and pathogenicity in pigs.

Development and evaluation of indirect enzyme-linked immunosorbent assay for a screening test to detect antibodies against classical swine fever virus
Yoshihiro Sakoda, Hiroaki Wakamoto, Tehpin Tamura, Takushi Nomura, Michiko Naito, Hiroshi Aoki, Hiroshi Morita, Hiroshi Kida, Akio Fukusho
JAPANESE JOURNAL OF VETERINARY RESEARCH, 60, 2-3, 85, 94, HOKKAIDO UNIV, 2012年08月, ［査読有り］
英語, 研究論文（学術雑誌）, An indirect enzyme-linked immunosorbent assay (ELISA) was developed for a screening test to detect antibodies against classical swine fever virus (CSFV). Viral glycoproteins, which were purified from swine kidney cells infected with CSFV ALD/A76 strain by the immunoaffinity purification using monoclonal antibody against E2 protein, were adsorbed on a microtiter plate as the antigen for the antibody detection. Each antibody titer of serum sample was expressed as a sample per positive value calculated with optical absorbance of each sample and that of a positive control. The advantage of this ELISA is its higher sensitivity: most sera containing more than 4 neutralization titers were determined to be positive. This ELISA is unable to discriminate between antibodies against CSFV and those against other ruminant pestiviruses, therefore positive sera in this ELISA should be evaluated by a cross-neutralization test using CSFV, bovine viral diarrhea virus, and border disease virus. Taken together, the indirect ELISA developed in this study is useful screening tool to detect antibodies against CSFV for the large-scale monitoring of classical swine fever.

Recovery of Leptospires from Miniature Pigs Experimentally Infected with Leptospira interrogans Serovar Manilae Strain UP-MMC under Immunosuppressive Conditions by Dexamethasone
Yoshihiro Sakoda, Michiko Naito, Mutsumi Ito, Yuki Ito, Norikazu Isoda, Tomohisa Tanaka, Takashi Umemura, Hiroshi Kida
JOURNAL OF VETERINARY MEDICAL SCIENCE, 74, 7, 955, 958, JAPAN SOC VET SCI, 2012年07月, ［査読有り］
英語, 研究論文（学術雑誌）, Leptospira interrogans serovar Manilae strain UP-MMC was inoculated into miniature pigs to assess its pathogenicity. Leptospires were recovered from the whole blood, kidneys, and livers in the acute phase without showing any clinical signs. Under immunosuppressive conditions by dexamethasone, leptospires were recovered from the kidneys and their genes were detected from the urine in the chronic phase. These results indicate that leptospires persisted in the kidneys until the chronic phase, and excretion of leptospires in the urine was enhanced under immunosuppressive conditions, resulting in horizontal transmission among pigs on farms.

Purification of human and avian influenza viruses using cellulose sulfate ester (Cellufine Sulfate) in the process of vaccine production
Yoshihiro Sakoda, Masatoshi Okamatsu, Norikazu Isoda, Naoki Yamamoto, Koichi Ozaki, Yasuto Umeda, Shigeyuki Aoyama, Hiroshi Kida
MICROBIOLOGY AND IMMUNOLOGY, 56, 7, 490, 495, WILEY-BLACKWELL, 2012年07月, ［査読有り］
英語, 研究論文（学術雑誌）, Affinity chromatography using sulfated, spherical cellulose beads (Cellufine Sulfate) was assessed for purification of influenza A and influenza B viruses. Recovery rates of viruses eluted from the beads were high for all tested virus strains. This method was also useful for removing chicken egg-derived impurities from allantoic fluids containing influenza viruses; the hemagglutination activity per amount of protein in the eluted sample was significantly higher than that in the applied sample. These results suggest that use of Cellufine Sulfate is a practical method for primary purification of influenza viruses in the process of influenza vaccine production.

Memory Immune Responses against Pandemic (H1N1) 2009 Influenza Virus Induced by a Whole Particle Vaccine in Cynomolgus Monkeys Carrying Mafa-A1*052:02
Masahiko Arikata, Yasushi Itoh, Masatoshi Okamatsu, Toshinaga Maeda, Takashi Shiina, Keiko Tanaka, Shingo Suzuki, Misako Nakayama, Yoshihiro Sakoda, Hirohito Ishigaki, Ayato Takada, Hideaki Ishida, Kosuke Soda, Van Loi Pham, Hideaki Tsuchiya, Shinichiro Nakamura, Ryuzo Torii, Takeshi Shimizu, Hidetoshi Inoko, Iwao Ohkubo, Hiroshi Kida, Kazumasa Ogasawara
PLOS ONE, 7, 5, e37220, PUBLIC LIBRARY SCIENCE, 2012年05月, ［査読有り］
英語, 研究論文（学術雑誌）, We made an H1N1 vaccine candidate from a virus library consisting of 144 (= 16 HAx9 NA) non-pathogenic influenza A viruses and examined its protective effects against a pandemic (2009) H1N1 strain using immunologically naive cynomolgus macaques to exclude preexisting immunity and to employ a preclinical study since preexisting immunity in humans previously vaccinated or infected with influenza virus might make comparison of vaccine efficacy difficult. Furthermore, macaques carrying a major histocompatibility complex class I molecule, Mafa-A1*052:02, were used to analyze peptide-specific CD8(+) T cell responses. Sera of macaques immunized with an inactivated whole particle formulation without addition of an adjuvant showed higher neutralization titers against the vaccine strain A/Hokkaido/2/1981 (H1N1) than did sera of macaques immunized with a split formulation. Neutralization activities against the pandemic strain A/Narita/1/2009 (H1N1) in sera of macaques immunized twice with the split vaccine reached levels similar to those in sera of macaques immunized once with the whole particle vaccine. After inoculation with the pandemic virus, the virus was detected in nasal samples of unvaccinated macaques for 6 days after infection and for 2.67 days and 5.33 days on average in macaques vaccinated with the whole particle vaccine and the split vaccine, respectively. After the challenge infection, recall neutralizing antibody responses against the pandemic virus and CD8(+) T cell responses specific for nucleoprotein peptide NP262-270 bound to Mafa-A1*052:02 in macaques vaccinated with the whole particle vaccine were observed more promptly or more vigorously than those in macaques vaccinated with the split vaccine. These findings demonstrated that the vaccine derived from our virus library was effective for pandemic virus infection in macaques and that the whole particle vaccine conferred more effective memory and broader cross-reactive immune responses to macaques against pandemic influenza virus infection than did the split vaccine.

An H9N2 Influenza Virus Vaccine Prepared from a Non-Pathogenic Isolate from a Migratory Duck Confers Protective Immunity in Mice against Challenge with an H9N2 Virus Isolated from a Girl in Hong Kong
Naoki Nomura, Yoshihiro Sakoda, Kosuke Soda, Masatoshi Okamatsu, Hiroshi Kida
JOURNAL OF VETERINARY MEDICAL SCIENCE, 74, 4, 441, 447, JAPAN SOC VET SCI, 2012年04月, ［査読有り］
英語, 研究論文（学術雑誌）, H9N2 influenza viruses circulate in wild birds and poultry in Eurasian countries, and have been isolated from pigs and humans in China. H9N2 viruses isolated from birds, pigs and humans have been classified into three sublineages based on antigenic and genetic features. Chicken antisera to H9N2 viruses of the Korean sublineage reacted with viruses of different sublineages by the hemagglutination-inhibition test. A test vaccine prepared from a non-pathogenic A/duck/Hokkaido/49/1998 (H9N2) strain of the Korean sublineage, obtained from our influenza virus library, induced immunity in mice to reduce the impact of disease caused by the challenge with A/Hong Kong/1073/1999 (H9N2), which is of a different sublineage. The present results indicate that an inactivated whole virus vaccine prepared from a non-pathogenic influenza virus from the library could be used as an emergency vaccine during the early stage of a pandemic caused by H9N2 infection.

カモから分離されたH7インフルエンザウイルスのニワトリに対する病原性獲得メカニズム　　　　　　　　　　　　　　　
丸山 隼輝, 曽田 公輔, 岡松 正敏, 迫田 義博, 喜田 宏
日本獣医学会学術集会講演要旨集, 153回, 229, 229, (公社)日本獣医学会, 2012年03月
日本語

Reintroduction of H5N1 highly pathogenic avian influenza virus by migratory water birds, causing poultry outbreaks in the 2010-2011 winter season in Japan
Yoshihiro Sakoda, Hiroshi Ito, Yuko Uchida, Masatoshi Okamatsu, Naoki Yamamoto, Kosuke Soda, Naoki Nomura, Saya Kuribayashi, Shintaro Shichinohe, Yuji Sunden, Takashi Umemura, Tatsufumi Usui, Hiroichi Ozaki, Tsuyoshi Yamaguchi, Toshiyuki Murase, Toshihiro Ito, Takehiko Saito, Ayato Takada, Hiroshi Kida
JOURNAL OF GENERAL VIROLOGY, 93, 3, 541, 550, SOC GENERAL MICROBIOLOGY, 2012年03月, ［査読有り］
英語, 研究論文（学術雑誌）, H5N1 highly pathogenic avian influenza virus (HPAIV) was reintroduced and caused outbreaks in chickens in the 2010-2011 winter season in Japan, which had been free from highly pathogenic avian influenza (HPAI) since 2007 when HPAI outbreaks occurred and were controlled. On 14 October 2010 at Lake Ohnuma, Wakkanai, the northernmost part of Hokkaido, Japan, H5N1 HPAIVs were isolated from faecal samples of ducks flying from their nesting lakes in Siberia. Since then, in Japan, H5N1 HPAIVs have been isolated from 63 wild birds in 17 prefectures and caused HPAI outbreaks in 24 chicken farms in nine prefectures by the end of March in 2011. Each of these isolates was genetically closely related to the HPAIV isolates at Lake Ohnuma, and those in China, Mongolia, Russia and Korea, belonging to genetic clade 2.3.2.1. In addition, these isolates were genetically classified into three groups, suggesting that the viruses were transmitted by migratory water birds through at least three different routes from their northern territory to Japan. These isolates were antigenic variants, which is consistent with selection in poultry under the immunological pressure induced by vaccination. To prevent the perpetuation of viruses in the lakes where water birds nest in summer in Siberia, prompt eradication of HPAIVs in poultry is urgently needed in Asian countries where HPAI has not been controlled.

Characterization of avian influenza viruses isolated from domestic ducks in Vietnam in 2009 and 2010
Naoki Nomura, Yoshihiro Sakoda, Mayumi Endo, Hiromi Yoshida, Naoki Yamamoto, Masatoshi Okamatsu, Kenji Sakurai, Nam Van Hoang, Long Van Nguyen, Huy Duc Chu, Tien Ngoc Tien, Hiroshi Kida
ARCHIVES OF VIROLOGY, 157, 2, 247, 257, SPRINGER WIEN, 2012年02月, ［査読有り］
英語, 研究論文（学術雑誌）, In the surveillance of avian influenza in Vietnam, 26 H9N2, 1 H3N2, 1 H3N8, 7 H4N6, 3 H11N3, and 1 H11N9 viruses were isolated from tracheal and cloacal swab samples of 300 domestic ducks in April 2009, and 1 H9N6 virus from 300 bird samples in March 2010. Out of the 27 H9 virus isolates, the hemagglutinins of 18 strains were genetically classified as belonging to the sublineage G1, and the other nine belonged to the Korean sublineage. Phylogenetic analysis revealed that one of the 27 H9 viruses was a reassortant in which the PB2 gene belonged to the Korean sublineage and the other seven genes belonged to the G1 sublineage. Three representative H9N2 viruses were intranasally inoculated into ducks, chickens, pigs, and mice. On the basis of experimental infection studies, it was found that each of the three viruses readily infected pigs and replicated in their upper respiratory tracts, and they infected chickens with slight replication. Viruses were recovered from the lungs of mice inoculated with two of the three isolates. The present results reveal that H9 avian influenza viruses are prevailing and genetic reassortment occurs among domestic ducks in Vietnam. It is recommended that careful surveillance of swine influenza with H9 viruses should be performed to prepare for pandemic influenza.

Adaptation of a Duck Influenza A Virus in Quail
Shinya Yamada, Kyoko Shinya, Ayato Takada, Toshihiro Ito, Takashi Suzuki, Yasuo Suzuki, Quynh Mai Le, Masahito Ebina, Noriyuki Kasai, Hiroshi Kida, Taisuke Horimoto, Pierre Rivailler, Li Mei Chen, Ruben O. Donis, Yoshihiro Kawaoka
JOURNAL OF VIROLOGY, 86, 3, 1411, 1420, AMER SOC MICROBIOLOGY, 2012年02月, ［査読有り］
英語, 研究論文（学術雑誌）, Quail are thought to serve as intermediate hosts of influenza A viruses between aquatic birds and terrestrial birds, such as chickens, due to their high susceptibility to aquatic-bird viruses, which then adapt to replicate efficiently in their new hosts. However, does replication of aquatic-bird influenza viruses in quail similarly result in their efficient replication in humans? Using sialic acid-galactose linkage-specific lectins, we found both avian (sialic acid-alpha 2-3-galactose [Sia alpha 2-3Gal] linkages on sialyloligosaccharides)- and human (Sia alpha 2-6Gal)-type receptors on the tracheal cells of quail, consistent with previous reports. We also passaged a duck H3N2 virus in quail 19 times. Sequence analysis revealed that eight mutations accumulated in hemagglutinin (HA) during these passages. Interestingly, many of the altered HA amino acids found in the adapted virus are present in human seasonal viruses, but not in duck viruses. We also found that stepwise stalk deletion of neuraminidase occurred during passages, resulting in reduced neuraminidase function. Despite some hemagglutinin mutations near the receptor binding pocket, appreciable changes in receptor specificity were not detected. However, reverse-genetics-generated viruses that possessed the hemagglutinin and neuraminidase of the quail-passaged virus replicated significantly better than the virus possessing the parent HA and neuraminidase in normal human bronchial epithelial cells, whereas no significant difference in replication between the two viruses was observed in duck cells. Further, the quail-passaged but not the original duck virus replicated in human bronchial epithelial cells. These data indicate that quail can serve as intermediate hosts for aquatic-bird influenza viruses to be transmitted to humans.

Osteopontin is critical to determine symptom severity of influenza through the regulation of NK cell population
Kayoko Sato, Atsushi Iwai, Yosuke Nakayama, Junko Morimoto, Ayato Takada, Mitsuo Maruyama, Hiroshi Kida, Toshimitsu Uede, Tadaaki Miyazaki
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 417, 1, 274, 279, ACADEMIC PRESS INC ELSEVIER SCIENCE, 2012年01月, ［査読有り］
英語, 研究論文（学術雑誌）, Osteopontin (OPN) is involved in exacerbating various inflammatory diseases. A severe pulmonary inflammation is frequently found in lethal influenza A virus (IAV) infection. However, the function of OPN against the infection was poorly understood. Here, we demonstrate an importance of OPN on immune response and disease severity after IAV infection. We found that the expression level of OPN was increased in mice infected with IAV. The OPN knockout (KO) mice exhibited a severe pathological phenotype and the survival rate decreased after the lethal IAV infection, compared to the wild type mice, while the survival rate increased in OPN transgenic (Tg) mice. The population of natural killer (NK) cells significantly decreased in OPN KO mice at day 5 after the infection, whereas, it increased in OPN Tg mice. These results suggest that OPN plays an important role in host defense against IAV infection through the regulation of NK cell population. (C) 2011 Elsevier Inc. All rights reserved.

Rapid typing of influenza viruses using super high-speed quantitative real-time PCR
Akira Sakurai, Namiko Nomura, Reiko Nanba, Takayuki Sinkai, Tsunehito Iwaki, Taminori Obayashi, Kazuhiro Hashimoto, Michiya Hasegawa, Yoshihiro Sakoda, Akihiro Naito, Yoshihito Morizane, Mitsugu Hosaka, Kunio Tsuboi, Hiroshi Kida, Akemi Kai, Futoshi Shibasaki
JOURNAL OF VIROLOGICAL METHODS, 178, 1-2, 75, 81, ELSEVIER SCIENCE BV, 2011年12月, ［査読有り］
英語, 研究論文（学術雑誌）, The development of a rapid and sensitive system for detecting influenza viruses is a high priority for controlling future epidemics and pandemics. Quantitative real-time PCR is often used for detecting various kinds of viruses; however, it requires more than 2 h per run. Detection assays were performed with super high-speed RT-PCR (SHRT-PCR) developed according to a newly designed heating system. The new method uses a high-speed reaction (18 s/cycle; 40 cycles in less than 20 min) for typing influenza viruses. The detection limit of SHRT-PCR was 1 copy/reaction and 10(-1) plaque-forming unit/reaction for viruses in culture supernatants during 20 min. Using SHRT-PCR, 86 strains of influenza viruses isolated by the Tokyo Metropolitan Institute of Public Health were tested; the results showed 100% sensitivity and specificity for each influenza A and B virus, and swine-origin influenza virus. Twenty-seven swabs collected from the pharyngeal mucosa of outpatients were also tested, showing positive signs for influenza virus on an immunochromatographic assay; the results between SHRT-PCR and immunochromatography exhibited 100% agreement for both positive and negative results. The rapid reaction time and high sensitivity of SHRT-PCR makes this technique well suited for monitoring epidemics and pre-pandemic influenza outbreaks. (C) 2011 Published by Elsevier B.V.

Development of a highly sensitive immunochromatographic detection kit for H5 influenza virus hemagglutinin using silver amplification
Atsuhiko Wada, Yoshihiro Sakoda, Takayoshi Oyamada, Hiroshi Kida
JOURNAL OF VIROLOGICAL METHODS, 178, 1-2, 82, 86, ELSEVIER SCIENCE BV, 2011年12月, ［査読有り］
英語, 研究論文（学術雑誌）, H5N1, a highly pathogenic avian influenza virus (HPAIV), has become a serious epizootic threat to the poultry population in Asia. In addition, significant numbers of human cases of HPAIV infection have been reported to date. To prevent the spread of HPAIV among humans and to allow for timely medical intervention, a rapid and high sensitive method is needed to detect and subtype the causative HPAIVs. In the present study, a silver amplification technique used in photographic development was combined with immunochromatography technologies and a highly sensitive and rapid diagnostic test to detect the hemagglutinin of H5 influenza viruses was developed. The sensitivity of the test kit was increased 500 times by silver amplification. The sensitivity of the method was more than 10 times higher than those of conventional rapid influenza diagnostic tests, which detect viral nucleoproteins. The diagnostic system developed in the present study can therefore provide rapid and highly sensitive results and will be useful for diagnosis of H5 HPAIV infection in humans and animals. (C) 2011 Elsevier B.V. All rights reserved.

Phase III Randomized, Double-Blind Study Comparing Single-Dose Intravenous Peramivir with Oral Oseltamivir in Patients with Seasonal Influenza Virus Infection
Shigeru Kohno, Muh-Yong Yen, Hee-Jin Cheong, Nobuo Hirotsu, Tadashi Ishida, Jun-ichi Kadota, Masashi Mizuguchi, Hiroshi Kida, Jingoro Shimada
ANTIMICROBIAL AGENTS AND CHEMOTHERAPY, 55, 11, 5267, 5276, AMER SOC MICROBIOLOGY, 2011年11月, ［査読有り］
英語, 研究論文（学術雑誌）, Antiviral medications with activity against influenza viruses are important in controlling influenza. We compared intravenous peramivir, a potent neuraminidase inhibitor, with oseltamivir in patients with seasonal influenza virus infection. In a multinational, multicenter, double-blind, double-dummy randomized controlled study, patients aged >= 20 years with influenza A or B virus infection were randomly assigned to receive either a single intravenous infusion of peramivir (300 or 600 mg) or oral administration of oseltamivir (75 mg twice a day [b.i.d.] for 5 days). To demonstrate the noninferiority of peramivir in reducing the time to alleviation of influenza symptoms with hazard model analysis and a noninferiority margin of 0.170, we planned to recruit 1,050 patients in South Korea, Japan, and Taiwan. A total of 1,091 patients (364 receiving 300 mg and 362 receiving 600 mg of peramivir; 365 receiving oseltamivir) were included in the intent-to-treat infected population. The median durations of influenza symptoms were 78.0, 81.0, and 81.8 h in the groups treated with 300 mg of peramivir, 600 mg of peramivir, and oseltamivir, respectively. The hazard ratios of the 300- and 600-mg-peramivir groups compared to the oseltamivir group were 0.946 (97.5% confidence interval [CI], 0.793, 1.129) and 0.970 (97.5% CI, 0.814, 1.157), respectively. Both peramivir groups were noninferior to the oseltamivir group (97.5% CI, <1.170). The overall incidence of adverse drug reactions was significantly lower in the 300-mg-peramivir group, but the incidence of severe reactions in either peramivir group was not different from that in the oseltamivir group. Thus, a single intravenous dose of peramivir may be an alternative to a 5-day oral dose of oseltamivir for patients with seasonal influenza virus infection.

Gnarled-Trunk Evolutionary Model of Influenza A Virus Hemagglutinin
Kimihito Ito, Manabu Igarashi, Yutaka Miyazaki, Teiji Murakami, Syaka Iida, Hiroshi Kida, Ayato Takada
PLOS ONE, 6, 10, e25953, PUBLIC LIBRARY SCIENCE, 2011年10月, ［査読有り］
英語, 研究論文（学術雑誌）, Human influenza A viruses undergo antigenic changes with gradual accumulation of amino acid substitutions on the hemagglutinin (HA) molecule. A strong antigenic mismatch between vaccine and epidemic strains often requires the replacement of influenza vaccines worldwide. To establish a practical model enabling us to predict the future direction of the influenza virus evolution, relative distances of amino acid sequences among past epidemic strains were analyzed by multidimensional scaling (MDS). We found that human influenza viruses have evolved along a gnarled evolutionary pathway with an approximately constant curvature in the MDS-constructed 3D space. The gnarled pathway indicated that evolution on the trunk favored multiple substitutions at the same amino acid positions on HA. The constant curvature was reasonably explained by assuming that the rate of amino acid substitutions varied from one position to another according to a gamma distribution. Furthermore, we utilized the estimated parameters of the gamma distribution to predict the amino acid substitutions on HA in subsequent years. Retrospective prediction tests for 12 years from 1997 to 2009 showed that 70% of actual amino acid substitutions were correctly predicted, and that 45% of predicted amino acid substitutions have been actually observed. Although it remains unsolved how to predict the exact timing of antigenic changes, the present results suggest that our model may have the potential to recognize emerging epidemic strains.

Antigenic and genetic analysis of H3N8 influenza viruses isolated from horses in Japan and Mongolia, and imported from Canada and Belgium during 2007-2010
Masayuki Motoshima, Masatoshi Okamatsu, Shingo Asakura, Saya Kuribayashi, Sugar Sengee, Damdinjav Batchuluun, Mika Ito, Yukiko Maeda, Mariko Eto, Yoshihiro Sakoda, Ruuragchaa Sodnomdarjaa, Hiroshi Kida
ARCHIVES OF VIROLOGY, 156, 8, 1379, 1385, SPRINGER WIEN, 2011年08月, ［査読有り］
英語, 研究論文（学術雑誌）, A/equine/Kanazawa/1/2007 (H3N8), A/equine/Hokkaido/I828/2008 (H3N8) and A/equine/Mongolia/1/2008 (H3N8) were isolated from infected horses. A/equine/Yokohama/aq19/2009 (H3N8) and A/equine/Yokohama/aq13/2010 (H3N8) were isolated from horses imported from Canada and Belgium examined at the Animal Quarantine Service in Yokohama, Japan. In the present study, these five isolates were genetically and antigenically analyzed. Phylogenetic analysis of hemagglutinin (HA) and neuraminidase (NA) genes showed that three isolates from horses in Japan and imported from Canada belonged to the same branch, clade 1 of the Florida sublineage, while the isolates from horses in Mongolia and imported from Belgium belonged to another branch, clade 2 of the Florida sublineage. Reactivity patterns of a panel of monoclonal antibodies to the HA of A/equine/Kanazawa/1/2007 (H3N8) with the five isolates indicate that the HAs of these viruses were antigenically similar to each other and to the reference strains A/equine/La Plata/1/1993 (H3N8) and A/equine/Avesta/1/1993 (H3N8). The present findings indicate that extensive antigenic variation has not accumulated among H3N8 influenza viruses in horses.

An H5N1 highly pathogenic avian influenza virus that invaded Japan through waterfowl migration
Masahiro Kajihara, Keita Matsuno, Edgar Simulundu, Mieko Muramatsu, Osamu Noyori, Rashid Manzoor, Eri Nakayama, Manabu Igarashi, Daisuke Tomabechi, Reiko Yoshida, Masatoshi Okamatsu, Yoshihiro Sakoda, Kimihito Ito, Hiroshi Kida, Ayato Takada
JAPANESE JOURNAL OF VETERINARY RESEARCH, 59, 2-3, 89, 100, HOKKAIDO UNIV, 2011年08月, ［査読有り］
英語, 研究論文（学術雑誌）, In 2010, an H5N1 highly pathogenic avian influenza virus (HPAIV) was isolated from feces of apparently healthy ducks migrating southward in Hokkaido, the northernmost prefecture of Japan. The H5N1 HPAIVs were subsequently detected in domestic and wild birds at multiple sites corresponding to the flyway of the waterfowl having stopovers in the Japanese archipelago. The Hokkaido isolate was genetically nearly identical to H5N1 HPAIVs isolated from swans in the spring of 2009 and 2010 in Mongolia, but less pathogenic in experimentally infected ducks than the 2009 Mongolian isolate. These findings suggest that H5N1 HPAIVs with relatively mild pathogenicity might be selected and harbored in the waterfowl population during the 2009-2010 migration seasons. Our data provide "early warning" signals for preparedness against the unprecedented situation in which the waterfowl reservoirs serve as perpetual sources and disseminators of HPAIVs.

Evaluation of the Reverse Transcription Loop-Mediated Isothermal Amplification (RT-LAMP) as a Screening Method for the Detection of Influenza Viruses in the Fecal Materials of Water Birds
Hiromi Yoshida, Yoshihiro Sakoda, Mayumi Endo, Masayuki Motoshima, Fumi Yoshino, Naoki Yamamoto, Masatoshi Okamatsu, Takahiro Soejima, Syouhei Senba, Hidetoshi Kanda, Hiroshi Kida
JOURNAL OF VETERINARY MEDICAL SCIENCE, 73, 6, 753, 758, JAPAN SOC VET SCI, 2011年06月, ［査読有り］
英語, 研究論文（学術雑誌）, Migratory water birds are a natural reservoir for influenza A viruses. Viruses replicate in the intestines of ducks and are shed with the fecal materials. Virus isolation from collected fecal materials, therefore, is an integral part of the surveillance of avian influenza in water birds. In the present study, reverse transcription loop-mediated isothermal amplification (RT-LAMP) was assessed for its usefulness in detecting the RNA of influenza A viruses in fecal materials. It was found that, RT-LAMP specifically and sensitively detects the matrix gene of influenza A viruses. Influenza A viruses were isolated from the fecal materials in which viral RNA were detected by RT-LAMP in 35 min. The present findings indicate that RI-LAMP is useful as a high throughput screening method for field samples prior to virus isolation, allowing the processing of hundreds of samples per day.

A Low Pathogenic H5N2 Influenza Virus Isolated in Taiwan Acquired High Pathogenicity by Consecutive Passages in Chickens
Kosuke Soda, Ming-Chu Cheng, Hiromi Yoshida, Mayumi Endo, Shu-Hwae Lee, Masatoshi Okamatsu, Yoshihiro Sakoda, Ching-Ho Wang, Hiroshi Kida
JOURNAL OF VETERINARY MEDICAL SCIENCE, 73, 6, 767, 772, JAPAN SOC VET SCI, 2011年06月, ［査読有り］
英語, 研究論文（学術雑誌）, H5N2 viruses were isolated from cloacal swab samples of apparently healthy chickens in Taiwan in 2003 and 2008 during surveillance of avian influenza. Each of the viruses was eradicated by stamping out. The official diagnosis report indicated that the Intravenous Pathogenicity Indexes (IVPIs) of the isolates were 0.00 and 0.89, respectively, indicating that these were low pathogenic strains, although the hemagglutinin of the strain isolated in 2008 (Taiwan08) had multibasic amino acid residues at the cleavage site (PQRKKR/G). In the present study, these H5N2 viruses were assessed for their intravenous and intranasal pathogenicity for chickens. It was examined whether Taiwan08 acquires pathogenicity through consecutive passages in chickens. Intravenous pathogenicity of Taiwan08 depended upon the age of the chickens used for the IVPI test; all of the eight-week-old chickens intravenously inoculated with Taiwan08 showed clinical signs but survived for ten days post inoculation (IVPI=0.68), whereas all the six-week-old chickens died (IVPI=1.86). Taiwan08-P8, which were passaged in chickens for eight times, killed all the eight-week-old chickens (IVPI=2.36). The four-week-old chickens died after intranasal inoculation of Taiwan08-P8, indicating that Taiwan08 must have become highly pathogenic during circulation in chicken flocks. These results emphasize the importance of a stamping out policy for avian influenza even if the IVPI of the causal virus is low.

Characterization of influenza A viruses isolated from wild waterfowl in Zambia
Edgar Simulundu, Akihiro Ishii, Manabu Igarashi, Aaron S. Mweene, Yuka Suzuki, Bernard M. Hang'ombe, Boniface Namangala, Ladslav Moonga, Rashid Manzoor, Kimihito Ito, Ichiro Nakamura, Hirofumi Sawa, Chihiro Sugimoto, Hiroshi Kida, Chuma Simukonda, Wilbroad Chansa, Jack Chulu, Ayato Takada
JOURNAL OF GENERAL VIROLOGY, 92, 6, 1416, 1427, SOC GENERAL MICROBIOLOGY, 2011年06月, ［査読有り］
英語, 研究論文（学術雑誌）, Although the quest to clarify the role of wild birds in the spread of the highly pathogenic H5N1 avian influenza virus (AIV) has yielded considerable data on AIVs in wild birds worldwide, information regarding the ecology and epidemiology of AIVs in African wild birds is still very limited. During AIV surveillance in Zambia (2008-2009), 12 viruses of distinct subtypes (H3N8, H4N6, H6N2, H9N1 and H11N9) were isolated from wild waterfowl. Phylogenetic analyses demonstrated that all the isolates were of the Eurasian lineage. Whilst some genes were closely related to those of AIVs isolated from wild and domestic birds in South Africa, intimating possible AIV exchange between wild birds and poultry in southern Africa, some gene segments were closely related to those of AIVs isolated in Europe and Asia, thus confirming the inter-regional AIV gene flow among these continents. Analysis of the deduced amino acid sequences of internal proteins revealed that several isolates harboured particular residues predominantly observed in human influenza viruses. Interestingly, the isolates with human-associated residues exhibited higher levels of virus replication in the lungs of infected mice and caused more morbidity as measured by weight loss than an isolate lacking such residues. This study stresses the need for continued monitoring of AIVs in wild and domestic birds in southern Africa to gain a better understanding of the emergence of strains with the potential to infect mammals.

Intravenous Peramivir for Treatment of Influenza A and B Virus Infection in High-Risk Patients
Shigeru Kohno, Hiroshi Kida, Masashi Mizuguchi, Nobuo Hirotsu, Tadashi Ishida, Junichi Kadota, Jingoro Shimada
ANTIMICROBIAL AGENTS AND CHEMOTHERAPY, 55, 6, 2803, 2812, AMER SOC MICROBIOLOGY, 2011年06月, ［査読有り］
英語, 研究論文（学術雑誌）, Influenza virus infections are known to persist longer in patients with underlying diseases, including respiratory tract diseases, and tend to become complicated by secondary influenza-associated infections, such as pneumonia. To assess the efficacy and safety of the novel anti-influenza virus drug peramivir in high-risk patients, we conducted a clinical trial of patients with diabetes or chronic respiratory tract diseases and patients being treated with drugs that suppress immune function. In this multicenter, uncontrolled, randomized, double-blind study, peramivir was intravenously administered at 300 or 600 mg/day for 1 to 5 days, as needed. Efficacy was investigated in 37 patients (300 mg, n = 18 patients; 600 mg, n = 19 patients). The median durations of influenza illness were 68.6 h (90% confidence interval, 41.5 to 113.4 h) overall, 114.4 h (90% confidence interval, 40.2 to 235.3 h) in the 300-mg group, and 42.3 h (90% confidence interval, 30.0 to 82.7 h) in the 600-mg group. The hazard ratio for the 600-mg group compared to the 300-mg group was 0.497 (90% confidence interval, 0.251 to 0.984), and the duration of influenza illness was significantly shorter in the 600-mg group than in the 300-mg group. Among the 42 patients in the safety analysis set, adverse events occurred in 73.8% and adverse drug reactions in 33.3%. No adverse events were particularly problematic clinically, and all patients recovered quickly from all events. The measured blood drug concentrations showed no tendency toward accumulation. Drug accumulation with repeated doses was thus considered to be of little concern. Intravenous peramivir appears to offer a potentially useful treatment for high-risk patients in the future.

Involvement of Endothelial Apoptosis Underlying Chronic Obstructive Pulmonary Disease-like Phenotype in Adiponectin-null Mice Implications for Therapy
Kaori Nakanishi, Yoshito Takeda, Satoshi Tetsumoto, Takeo Iwasaki, Kazuyuki Tsujino, Hanako Kuhara, Yingji Jin, Izumi Nagatomo, Hiroshi Kida, Sho Goya, Takashi Kijima, Norikazu Maeda, Tohru Funahashi, Iichiro Shimomura, Isao Tachibana, Ichiro Kawase
AMERICAN JOURNAL OF RESPIRATORY AND CRITICAL CARE MEDICINE, 183, 9, 1164, 1175, AMER THORACIC SOC, 2011年05月, ［査読有り］
英語, 研究論文（学術雑誌）, Rationale: Chronic obstructive pulmonary disease is frequently complicated with comorbidities, such as cardiovascular disease, osteoporosis, and body weight loss, but the causal link remains unclear.
Objectives: To investigate the role of adiponectin in the pathogenesis of chronic obstructive pulmonary disease and its potential use in therapy.
Methods: Adiponectin localization and dynamics in the lung were analyzed in an elastase-induced emphysema model. Next, the lung of adiponectin-knockout mice, extrapulmonary effects, and the underlying mechanism were investigated. Finally, we tested whether exogenous adiponectin could ameliorate the emphysematous change in adiponectin-knockout mice.
Measurements and Main Results: Adiponectin expression in lung vasculature and plasma concentration of adiponectin were reduced after elastase-instillation. Notably, adiponectin-knockout mice showed progressive alveolar enlargement and increased lung compliance. They further exhibited not only systemic inflammation, but also extrapulmonary phenotype, such as body weight loss, fat atrophy, and osteoporosis. Moreover, endothelial apoptosis was enhanced in the lungs of adiponectin-knockout mice, as evidenced by caspase-3 activity. Consistent with this, expressions of vascular endothelial growth factor receptor-2 and platelet endothelial cell adhesion molecule-1 on endothelial cells were decreased in the adiponectin-knockout mice. Finally, adenovirus-mediated adiponectin supplementation ameliorated the emphysematous phenotype.
Conclusions: Adiponectin-knockout mice develop progressive chronic obstructive pulmonary disease-like phenotype with systemic inflammation and extrapulmonary phenotypes. Hypoadiponectinemia could thus play a critical role in the progression of chronic obstructive pulmonary disease and concomitant comorbidities through endothelial dysfunction. Together, adiponectin could be a novel target for chronic obstructive pulmonary disease therapy.

Development of a novel rapid immunochromatographic test specific for the H5 influenza virus
Eiji Miyagawa, Hiroyuki Kogaki, Yoshiaki Uchida, Nobuyuki Fujii, Takashi Shirakawa, Yoshihrio Sakoda, Hiroshi Kida
JOURNAL OF VIROLOGICAL METHODS, 173, 2, 213, 219, ELSEVIER SCIENCE BV, 2011年05月, ［査読有り］
英語, 研究論文（学術雑誌）, Three anti-H5 influenza virus monoclonal antibody (mAb) clones, IFH5-26, IFH5-115 and IFH5-136, were obtained by immunising a BALB/C mouse with inactivated A/duck/Hokkaido/Vac-1/04 (H5N1). These mAbs were found to recognise specifically the haemagglutinin (HA) epitope of the influenza H5 subtypes by western blotting with recombinant HAs; however, these mAbs have no neutralising activity for A/duck/Hokkaido/84/02 (H5N3) or A/Puerto Ric/8/34 (H1N1). Each epitope of these mAbs was a conformational epitope that was formed from the regions located between 46 to 60 amino acids (aa) and 312 to 322 aa for IFH5-115, from 101 to 113 aa and 268 to 273 aa for IFH5-136 and from 61 to 80 aa and 290 to 300 aa for IFH5-26. The epitopes were located in the loop regions between the receptor region and alpha-helix structure in haemagglutinin 1 (HA1). Influenza A virus H5-specific rapid immunochromatographic test kits were tested as solid phase antibody/alkaline phosphate-conjugated mAb in the following three combinations: IFH5-26/IFH5-115, IFH5-136/IFH5-26 and IFH5-136/IFH5-115. In every combination, only influenza A H5 subtypes were detected. For effective clinical application, rapid dual discrimination immunochromatographic test kits in combination with H5 HA-specific mAb, IFA5-26 and IFA5-115 and the influenza A NP NP-specific mAb, FVA2-11, were developed. The dual discrimination immunochromatographic tests kits detected influenza A virus H5 subtypes as H5 line-positive and all influenza A subtypes as A line-positive simultaneously. The dual discrimination immunochromatographic test kits may be useful for discriminating highly pathogenic avian influenza A H5N1 viruses from seasonal influenza A virus, as well as for confirming influenza infection status in human, avian and mammalian hosts. (C) 2011 Elsevier B.V. All rights reserved.

H9N2 influenza virus vaccine prepared from a non-pathogenic isolate from a natural reservoir conferred protective immunity against the challenge with a human H9N2 virus in mice
Naoki Nomura, Yoshihiro Sakoda, Kosuke Soda, Masatoshi Okamatsu, Hiroshi Kida
INFLUENZA AND OTHER RESPIRATORY VIRUSES, 5, 363, 366, WILEY-BLACKWELL, 2011年05月, ［査読有り］
英語, 研究論文（学術雑誌）

H9N2 influenza virus acquires intravenous pathogenicity on the introduction of a pair of di-basic amino acid residues at the cleavage site of the hemagglutinin and consecutive passages in chickens
Kosuke Soda, Shingo Asakura, Masatoshi Okamatsu, Yoshihiro Sakoda, Hiroshi Kida
INFLUENZA AND OTHER RESPIRATORY VIRUSES, 5, 272, 276, WILEY-BLACKWELL, 2011年05月, ［査読有り］
英語, 研究論文（学術雑誌）

Prediction of the antigenic changes of the pandemic (H1N1) 2009 influenza virus hemagglutinin
Manabu Igarashi, Kimihito Ito, Reiko Yoshida, Daisuke Tomabechi, Hiroshi Kida, Ayato Takada
INFLUENZA AND OTHER RESPIRATORY VIRUSES, 5, 402, 404, WILEY-BLACKWELL, 2011年05月, ［査読有り］
英語, 研究論文（学術雑誌）

Improvement of the H5N1 influenza virus vaccine strain to decrease the pathogenicity in chicken embryos
Norikazu Isoda, Yoshihiro Sakoda, Masatoshi Okamatsu, Yoshimi Tsuda, Hiroshi Kida
ARCHIVES OF VIROLOGY, 156, 4, 557, 563, SPRINGER WIEN, 2011年04月, ［査読有り］
英語, 研究論文（学術雑誌）, The avian influenza vaccine strain A/duck/Hokkaido/Vac-1/2004 (H5N1) (Vac-1) was found to be pathogenic in chicken embryos (CEs). In order to decrease the pathogenicity of Vac-1 in CEs, a series of reassortant viruses was generated between Vac-1 and A/Puerto Rico/8/1934 (H1N1) (PR8), and their pathogenicity and growth potential were compared in CEs. The results indicated that either the PB1 or PA protein was responsible for the pathogenicity of Vac-1 in CEs. The HA titers of the allantoic fluids of CEs inoculated with the recombinant H5N1 viruses, of which pathogenicity was lower than that of the recombinant Vac-1 prepared by reverse genetics in CEs, were equivalent to those of CEs inoculated with the recombinant Vac-1. One of the reassortant viruses, rg-PR8-PA/Vac-1 (H5N1), in which the PA gene was replaced with the corresponding gene of PR8, yielded allantoic fluids with the same HA titer as that of Vac-1, indicating that this reassortant should be a good candidate as an improved vaccine strain.

Requirement for Siva-1 for replication of influenza A virus through apoptosis induction
Takuya Shiozaki, Atsushi Iwai, Yoshihiro Kawaoka, Ayato Takada, Hiroshi Kida, Tadaaki Miyazaki
JOURNAL OF GENERAL VIROLOGY, 92, 2, 315, 325, SOC GENERAL MICROBIOLOGY, 2011年02月, ［査読有り］
英語, 研究論文（学術雑誌）, Infection with influenza A virus causes acute respiratory tract infections in humans and may lead to lethal diseases including pneumonia. Identifying host factors that are involved in the severity of infectious diseases caused by influenza A virus is considered important for the prevention and treatment of these viral infections. This report demonstrated that Siva-1 is crucial for the induction of apoptosis caused by infection with influenza A virus and is involved in virus replication. Susceptibility to apoptosis induced by influenza A virus infection was increased in human lung-derived A549 cells, which stably express Siva-1. In addition, induction of apoptosis after influenza A virus infection was strongly inhibited by knockdown of Siva-1 expression. Furthermore, the replication of influenza A virus was significantly suppressed in A549 cells in which Siva-1 expression was inhibited and the effect of Siva-1 knockdown was eliminated by treatment with Z-VAD-FMK. These findings suggest that the caspase-dependent pathway for induction of apoptosis is involved in Siva-1-mediated influenza A virus replication.

H9N2 influenza virus acquires intravenous pathogenicity on the introduction of a pair of di-basic amino acid residues at the cleavage site of the hemagglutinin and consecutive passages in chickens
Kosuke Soda, Shingo Asakura, Masatoshi Okamatsu, Yoshihiro Sakoda, Hiroshi Kida
VIROLOGY JOURNAL, 8, 64, BIOMED CENTRAL LTD, 2011年02月, ［査読有り］
英語, 研究論文（学術雑誌）, Background: Outbreaks of avian influenza (AI) caused by infection with low pathogenic H9N2 viruses have occurred in poultry, resulting in serious economic losses in Asia and the Middle East. It has been difficult to eradicate the H9N2 virus because of its low pathogenicity, frequently causing in apparent infection. It is important for the control of AI to assess whether the H9N2 virus acquires pathogenicity as H5 and H7 viruses. In the present study, we investigated whether a non-pathogenic H9N2 virus, A/chicken/Yokohama/aq-55/2001 (Y55) (H9N2), acquires pathogenicity in chickens when a pair of di-basic amino acid residues is introduced at the cleavage site of its HA molecule.
Results: rgY55sub (H9N2), which had four basic amino acid residues at the HA cleavage site, replicated in MDCK cells in the absence of trypsin after six consecutive passages in the air sacs of chicks, and acquired intravenous pathogenicity to chicken after four additional passages. More than 75% of chickens inoculated intravenously with the passaged virus, rgY55sub-P10 (H9N2), died, indicating that it is pathogenic comparable to that of highly pathogenic avian influenza viruses (HPAIVs) defined by World Organization for Animal Health (OIE). The chickens inoculated with the virus via the intranasal route, however, survived without showing any clinical signs. On the other hand, an avirulent H5N1 strain, A/duck/Hokkaido/Vac-1/2004 (Vac1) (H5N1), acquired intranasal pathogenicity after a pair of di-basic amino acid residues was introduced into the cleavage site of the HA, followed by two passages by air sac inoculation in chicks.
Conclusion: The present results demonstrate that an H9N2 virus has the potential to acquire intravenous pathogenicity in chickens although the morbidity via the nasal route of infection is lower than that of H5N1 HPAIV.

Characterization of a non-pathogenic H5N1 influenza virus isolated from a migratory duck flying from Siberia in Hokkaido, Japan, in October 2009
Naoki Yamamoto, Yoshihiro Sakoda, Masayuki Motoshima, Fumi Yoshino, Kosuke Soda, Masatoshi Okamatsu, Hiroshi Kida
VIROLOGY JOURNAL, 8, 65, BIOMED CENTRAL LTD, 2011年02月, ［査読有り］
英語, 研究論文（学術雑誌）, Background: Infection with H5N1 highly pathogenic avian influenza viruses (HPAIVs) of domestic poultry and wild birds has spread to more than 60 countries in Eurasia and Africa. It is concerned that HPAIVs may be perpetuated in the lakes in Siberia where migratory water birds nest in summer. To monitor whether HPAIVs circulate in migratory water birds, intensive surveillance of avian influenza has been performed in Mongolia and Japan in autumn each year. Until 2008, there had not been any H5N1 viruses isolated from migratory water birds that flew from their nesting lakes in Siberia. In autumn 2009, A/mallard/Hokkaido/24/09 (H5N1) (Mal/Hok/24/09) was isolated from a fecal sample of a mallard (Anas platyrhynchos) that flew from Siberia to Hokkaido, Japan. The isolate was assessed for pathogenicity in chickens, domestic ducks, and quails and analyzed antigenically and phylogenetically.
Results: No clinical signs were observed in chickens inoculated intravenously with Mal/Hok/24/09 (H5N1). There was no viral replication in chickens inoculated intranasally with the isolate. None of the domestic ducks and quails inoculated intranasally with the isolate showed any clinical signs. There were no multiple basic amino acid residues at the cleavage site of the hemagglutinin (HA) of the isolate. Each gene of Mal/Hok/24/09 (H5N1) is phylogenetically closely related to that of influenza viruses isolated from migratory water birds that flew from their nesting lakes in autumn. Additionally, the antigenicity of the HA of the isolate was similar to that of the viruses isolated from migratory water birds in Hokkaido that flew from their northern territory in autumn and different from those of HPAIVs isolated from birds found dead in China, Mongolia, and Japan on the way back to their northern territory in spring.
Conclusion: Mal/Hok/24/09 (H5N1) is a non-pathogenic avian influenza virus for chickens, domestic ducks, and quails, and is antigenically and genetically distinct from the H5N1 HPAIVs prevailing in birds in Eurasia and Africa. H5 viruses with the HA gene of HPAIV had not been isolated from migratory water birds in the surveillance until 2009, indicating that H5N1 HPAIVs had not become dominant in their nesting lakes in Siberia until 2009.

Virological surveillance and phylogenetic analysis of the PB2 genes of influenza viruses isolated from wild water birds flying from their nesting lakes in Siberia to Hokkaido, Japan in autumn
Rozanah Asmah Abdul Samad, Yoshihiro Sakoda, Yoshimi Tsuda, Edgar Simulundu, Rashid Manzoor, Masatoshi Okamatsu, Kimihito Ito, Hiroshi Kida
JAPANESE JOURNAL OF VETERINARY RESEARCH, 59, 1, 15, 22, HOKKAIDO UNIV, 2011年02月, ［査読有り］
英語, 研究論文（学術雑誌）, Recent introduction of H5N1 highly pathogenic avian influenza virus (HPAIV) in wild birds from poultry in Eurasia signaled the possibility that this virus may perpetuate in nature. Surveillance of avian influenza especially in migratory birds, therefore, has been conducted to provide information on the viruses brought by them to Hokkaido, Japan, from their nesting lakes in Siberia in autumn. During 2008-2009, 62 influenza viruses of 21 different combinations of hemagglutinin (HA) and neuraminidase (NA) subtypes were isolated. Up to September 2010, no HPAIV has been found, indicating that H5N1 HPAIV has not perpetuated at least dominantly in the lakes where ducks nest in summer in Siberia. The PB2 genes of 54 influenza viruses out of 283 influenza viruses isolated in Hokkaido in 2000-2009 were phylogenetically analysed. None of the genes showed close relation to those of H5N1 HPAIVs that were detected in wild birds found dead in Eurasia on the way back to their northern territory in spring.

A vaccine prepared from a non-pathogenic H5N1 influenza virus strain from the influenza virus library conferred protective immunity to chickens against the challenge with antigenically drifted highly pathogenic avian influenza virus
Rozanah Asmah Abdul Samad, Naoki Nomura, Yoshimi Tsuda, Rashid Manzoor, Masahiro Kajihara, Daisuke Tomabechi, Takashi Sasaki, Norihide Kokumai, Toshiaki Ohgitani, Masatoshi Okamatsu, Ayato Takada, Yoshihiro Sakoda, Hiroshi Kida
JAPANESE JOURNAL OF VETERINARY RESEARCH, 59, 1, 23, 29, HOKKAIDO UNIV, 2011年02月, ［査読有り］
英語, 研究論文（学術雑誌）, Inactivated influenza virus vaccine prepared from a non-pathogenic influenza virus strain A/duck/Hokkaido/Vac-1/2004 (H5N1) from the virus library conferred protective immunity to chickens against the challenge of antigenically drifted highly pathogenic avian influenza virus (HPAIV), A/whooper swan/Hokkaido/1/2008 (H5N1). The efficacy of the vaccine was comparable to that prepared from genetically modified HPAIV strain Delta RRRRK rg-A/whooper swan/Mongolia/3/2005 (H5N1), which is more antigenically related to the challenge virus strain, in chickens.

Detection of highly pathogenic avian influenza virus infection in vaccinated chicken flocks by monitoring antibodies against non-structural protein 1 (NS1)
Natsumi Takeyama, Kenji Minari, Masahiro Kajihara, Norikazu Isoda, Ryuichi Sakamoto, Takashi Sasaki, Norihide Kokumai, Noriyasu Takikawa, Rikiya Shiraishi, Masaji Mase, Junko Hagiwara, Toshiaki Kodama, Takashi Imamura, Masashi Sakaguchi, Toshiaki Ohgitani, Akira Sawata, Masatoshi Okamatsu, Masatake Muramatsu, Kenji Tsukamoto, Zhifeng Lin, Kotaro Tuchiya, Yoshihiro Sakoda, Hiroshi Kida
VETERINARY MICROBIOLOGY, 147, 3-4, 283, 291, ELSEVIER SCIENCE BV, 2011年01月, ［査読有り］
英語, 研究論文（学術雑誌）, H5 and H7 highly pathogenic avian influenza virus (HPAIV) represent a major global concern in poultries and human health. Avian influenza (AI) vaccines are available but not preferred for field applications, primarily because vaccination interferes with sero-surveillances of AIV infection. To overcome the problem, ELISA systems using nonstructural protein 1 (NS1) of AIV as antigens (NS1-ELISA) have been developed to measure anti-NS1 antibodies that are raised in AIV-infected but not in vaccinated chickens. However, some AI-vaccinated chickens having a weak anti-virus immune response may subsequently be infected with AIV and spread the virus. This raises a concern for the validity of NS1-ELISA to detect AIV infection in previously vaccinated chickens. In this study, we developed NS1-ELISA and assessed its feasibility to detect HPAIV infection in chickens previously immunized with H5 or H7 AI vaccines. The results indicated that the NS1-ELISA could identify HPAIV infection in both unvaccinated and vaccinated chickens at 1 week after infection in correlation with results from time-consuming virus isolation tests. Taken together, the NS1-ELISA system would be valuable tool to define HPAIV infection when AI vaccine program is in place. (C) 2010 Elsevier B.V. All rights reserved.

Genetic characterization and susceptibility on poultry and mammal of H7N6 subtype avian influenza virus isolated in Japan in 2009
Yuko Uchida, Katsushi Kanehira, Masaji Mase, Nobuhiro Takemae, Chiaki Watanabe, Tatsufumi Usui, Yoshikazu Fujimoto, Toshihiro Ito, Manabu Igarashi, Kimihito Ito, Ayato Takada, Yoshihiro Sakoda, Masatoshi Okamatsu, Yu Yamamoto, Kikuyasu Nakamura, Hiroshi Kida, Yasuaki Hiromoto, Tomoyuki Tsuda, Takehiko Saito
VETERINARY MICROBIOLOGY, 147, 1-2, 1, 10, ELSEVIER SCIENCE BV, 2011年01月, ［査読有り］
英語, 研究論文（学術雑誌）, From February to March 2009, six strains of H7N6 subtype avian influenza virus were isolated from quails in three farms in Aichi prefecture in Japan. The isolates were shown to be low pathogenic for chicken by the examination performed using the "Manual of Standards for Diagnostic Tests and Vaccines" by World organisation for Animal Health (OIE). The deduced amino acid sequence at the cleavage site was PE (I/Q/L) PKRR (nucleotide sequences were cct gaa (a/c) (t/a) a cc (a/g) aaa aga aga), suggesting persistence in domestic poultry for some time. The direct putative ancestor strain could not be elucidated by phylogenetic analysis of all genome segments of the quail isolates. Diverged date from a putative common ancestor in a non-rooted phylogenetic tree among quail viruses was estimated between March 2002 and July 2004. Three putative N-linked glycosylation sites resided in the vicinity of the receptor binding pocket of HA1 region. They are considered to decrease the reactivity of neutralizing antibody against the virus. Experiments for the infectivity and pathogenicity of a quail strain to poultry indicated that the quail isolate had higher infectivity to quails than chickens and ducks. Direct and dust-borne and/or droplet-borne transmissions among quail were proven in quails with and without direct contact with experimentally infected quails. The virus is seldom transmitted among chickens either directly or indirectly, and indirect transmission from infected quails to chickens was not observed. The pathogenicity of the quail strain for mammalian, pig and mouse was low, although it could replicate in those animals. (C) 2010 Elsevier B.V. All rights reserved.

Diastereoselective synthesis of the indeno-tetrahydropyridine core bearing a diaryl-substituted stereogenic quaternary carbon center of haouamine B
Takeshi Tanaka, Hiromi Inui, Hiroshi Kida, Takeshi Kodama, Takuya Okamoto, Aki Takeshima, Yoshimitsu Tachi, Yoshiki Morimoto
CHEMICAL COMMUNICATIONS, 47, 10, 2949, 2951, ROYAL SOC CHEMISTRY, 2011年, ［査読有り］
英語, 研究論文（学術雑誌）, The characteristic indeno-tetrahydropyridine core of cytotoxic haouamine B (2) was efficiently synthesized featuring the diastereoselective construction of a diaryl-substituted stereogenic quaternary center by an intramolecular Pd-catalyzed alpha-C-arylation and subsequent direct conversion of the vinylogous imide function into the C2-C25 double bond by TsNHNH(2).

Antigenic, genetic, and pathogenic characterization of H5N1 highly pathogenic avian influenza viruses isolated from dead whooper swans (Cygnus cygnus) found in northern Japan in 2008
Masatoshi Okamatsu, Tomohisa Tanaka, Naoki Yamamoto, Yoshihiro Sakoda, Takashi Sasaki, Yoshimi Tsuda, Norikazu Isoda, Norihide Kokumai, Ayato Takada, Takashi Umemura, Hiroshi Kida
VIRUS GENES, 41, 3, 351, 357, SPRINGER, 2010年12月, ［査読有り］
英語, 研究論文（学術雑誌）, In April and May 2008, whooper swans (Cygnus cygnus) were found dead in Hokkaido in Japan. In this study, an adult whooper swan found dead beside Lake Saroma was pathologically examined and the identified H5N1 influenza virus isolates were genetically and antigenically analyzed. Pathological findings indicate that the swan died of severe congestive edema in the lungs. Phylogenetic analysis of the HA genes of the isolates revealed that they are the progeny viruses of isolates from poultry and wild birds in China, Russia, Korea, and Hong Kong. Antigenic analyses indicated that the viruses are distinguished from the H5N1 viruses isolated from wild birds and poultry before 2007. The chickens vaccinated with A/duck/Hokkaido/Vac-1/2004 (H5N1) survived for 14 days after challenge with A/whooper swan/Hokkaido/1/2008 (H5N1), although a small amount of the challenge virus was recovered from the tissues of the birds. These findings indicate that H5N1 highly pathogenic avian influenza viruses are circulating in wild birds in addition to domestic poultry in Asia and exhibit antigenic variation that may be due to vaccination.

Efficacy and Safety of Intravenous Peramivir for Treatment of Seasonal Influenza Virus Infection
Shigeru Kohno, Hiroshi Kida, Masashi Mizuguchi, Jingoro Shimada
ANTIMICROBIAL AGENTS AND CHEMOTHERAPY, 54, 11, 4568, 4574, AMER SOC MICROBIOLOGY, 2010年11月, ［査読有り］
英語, 研究論文（学術雑誌）, Peramivir, a sialic acid analogue, is a selective inhibitor of neuraminidases produced by influenza A and B viruses. We evaluated the efficacy and safety of a single intravenous dose of peramivir in outpatients with uncomplicated seasonal influenza virus infection. A total of 300 previously healthy adult subjects aged 20 to 64 years with a positive influenza virus rapid antigen test were recruited within 48 h of the onset of influenza symptoms and randomized to three groups: single intravenous infusion of either 300 mg peramivir per kg of body weight, 600 mg peramivir, or matching placebo on study day 1. Influenza symptoms and body temperature were self-assessed for 14 days. Nasal and pharyngeal swabs were collected to determine the viral titer. The primary endpoint was the time to alleviation of symptoms. Of the 300 subjects, 296 were included in the intent-to-treat infected population (300 mg peramivir, n = 99; 600 mg peramivir, n = 97; and placebo, n = 100). Peramivir significantly reduced the time to alleviation of symptoms at both 300 mg (hazard ratio, 0.681) and 600 mg (hazard ratio, 0.666) compared with placebo (adjusted P value, 0.0092 for both comparisons). No serious adverse events were reported. Peramivir was well tolerated, and its adverse-event profile was similar to that of placebo. A single intravenous dose of peramivir is effective and well tolerated in subjects with uncomplicated seasonal influenza virus infection.

Characterization of H5N1 highly pathogenic avian influenza virus strains isolated from migratory waterfowl in Mongolia on the way back from the southern Asia to their northern territory
Yoshihiro Sakoda, Sengee Sugar, Damdinjav Batchluun, Tseren-Ochir Erdene-Ochir, Masatoshi Okamatsu, Norikazu Isoda, Kosuke Soda, Hiroki Takakuwa, Yoshimi Tsuda, Naoki Yamamoto, Noriko Kishida, Keita Matsuno, Eri Nakayama, Masahiro Kajihara, Ayaka Yokoyama, Ayato Takada, Ruuragchaa Sodnomdarjaa, Hiroshi Kida
VIROLOGY, 406, 1, 88, 94, ACADEMIC PRESS INC ELSEVIER SCIENCE, 2010年10月, ［査読有り］
英語, 研究論文（学術雑誌）, H5N1 highly pathogenic avian influenza (HPAI) viruses were isolated from dead wild waterfowl at Khunt, Erkhel, Doityn Tsagaan, Doroo, and Ganga Lakes in Mongolia in July 2005, May 2006, May 2009, July 2009, and May 2010, respectively. The isolates in 2005 and 2006 were classified into genetic clade 2.2, and those in 2009 and 2010 into clade 2.3.2. A/whooper swan/Mongolia/6/2009 (H5N1) experimentally infected ducks and replicated systemically with higher mortality than that of the isolates in 2005 and 2006. Intensive surveillance of avian influenza in migratory waterfowl flying from their nesting lakes in Siberia to Mongolia in every autumn indicate that HPAI viruses have not perpetuated at their nesting lakes until 2009. The present results demonstrate that wild waterfowl were sporadically infected with H5N1 HPAI viruses prevailing in domestic poultry in the southern Asia and died in Mongolia on the way back to their northern territory in spring. (C) 2010 Elsevier Inc. All rights reserved.

Influenza A Virus Polymerase Inhibits Type I Interferon Induction by Binding to Interferon beta Promoter Stimulator 1
Atsushi Iwai, Takuya Shiozaki, Taro Kawai, Shizuo Akira, Yoshihiro Kawaoka, Ayato Takada, Hiroshi Kida, Tadaaki Miyazaki
JOURNAL OF BIOLOGICAL CHEMISTRY, 285, 42, 32064, 32074, AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC, 2010年10月, ［査読有り］
英語, 研究論文（学術雑誌）, Type I interferons (IFNs) are known to be critical factors in the activation of host antiviral responses and are also important in protection from influenza A virus infection. Especially, the RIG-I- and IPS-1-mediated intracellular type I IFN-inducing pathway is essential in the activation of antiviral responses in cells infected by influenza A virus. Previously, it has been reported that influenza A virus NS1 is involved in the inhibition of this pathway. We show in this report that the influenza A virus utilizes another critical inhibitory mechanism in this pathway. In fact, the viral polymerase complex exhibited an inhibitory activity on IFN beta promoter activation mediated by RIG-I and IPS-1, and this activity was not competitive with the function of NS1. Co-immunoprecipitation analysis revealed that each polymerase subunit bound to IPS-1 in mammalian cells, and each subunit inhibited the activation of IFN beta promoter by IPS-1 independently. In addition, by a combinational expression of each polymerase subunit, IPS-1-induced activation of IFN beta promoter was more efficiently inhibited by the expression of PB2 or PB2-containing complex. Moreover, the expression of PB2 inhibited the transcription of the endogenous IFN beta gene induced after influenza A virus infection. These findings demonstrate that the viral polymerase plays an important role for regulating host anti-viral response through the binding to IPS-1 and inhibition of IFN beta production.

A Comparison of the Antibody Responses between Specific Pathogen-Free and Commercial Layers Immunized with an Influenza Vaccine Prepared from Inactivated Non-Pathogenic H5N1 Virus by Single Shot
Takashi Sasaki, Norihide Kokumai, Toshiaki Ohgitani, Takashi Imamura, Akira Sawata, Zhifeng Lin, Yoshihiro Sakoda, Hiroshi Kida
JOURNAL OF VETERINARY MEDICAL SCIENCE, 72, 6, 819, 821, JAPAN SOC VET SCI, 2010年06月, ［査読有り］
英語, 研究論文（学術雑誌）, It is known that antibody responses in chickens against invading organisms or antigens are considerably different among different lines. Thus, an avian influenza vaccine was prepared from inactivated whole particles of the virus of non-pathogenic strain A/duck/Hokkaido/Vac-1/04 (H5N1) using an oil adjuvant containing anhydromannitol-octadecenoate-ether and injected intramuscularly into each ten 10-week-old specific pathogen-free (SPF) white leghorn chickens and commercial layers of Julia and Boris-Brown to obtain comparative data for antibody responses until 6 weeks after vaccination. Despite significant partial differences of antibody titer between the chicken lines, this study clearly showed that the vaccine induced good and sufficient antibody response in both SPF chickens and commercial layers.

Novel trivalent anti-influenza reagent
Fei Feng, Nobuaki Miura, Norikazu Isoda, Yoshihiro Sakoda, Masatoshi Okamatsu, Hiroshi Kida, Shin-Ichiro Nishimura
BIOORGANIC & MEDICINAL CHEMISTRY LETTERS, 20, 12, 3772, 3776, PERGAMON-ELSEVIER SCIENCE LTD, 2010年06月, ［査読有り］
英語, 研究論文（学術雑誌）, We designed and synthesized novel trivalent anti-influenza reagents. Sialyllactose was located at the terminal of each valence which aimed to block each receptor-binding site of the hemagglutinin (HA) trimer on the surface of the virus. Structural analyses were carried out with a model which was constructed with a computer simulation. A previously reported cyclic glycopeptide blocker [Ohta, T.; Miura, N.; Fujitani, N.; Nakajima, F.; Niikura, K.; Sadamoto, R.; Guo, C.-T.; Suzuki, T.; Suzuki, Y.; Monde, K.; Nishimura, S.-I. Angew. Chem. Int. Ed., 2003, 42, 5186] bound to the HA in the model. The analyses suggest that the glutamine residue in the cyclic peptide bearing Neu5A alpha 2,3Gal beta 1,4Glc trisaccharide via a linker interacts with the Gln189 in HA through hydrogen bonding. The present anti-influenza reagents likely interact with a glutamine residue included in the vicinity of Gln189. A plague reduction assay of the influenza virus, A/PR/8/1934 (H1N1),was performed in MDCK cells to evaluate for the synthesized compounds to inhibit viral replication. One of the compounds showed approximately 85% inhibition at the concentration of 400 mu M at 4 degrees C. (C) 2010 Elsevier Ltd. All rights reserved.

Isolation and Characterization of Potentially Pathogenic H5N2 Influenza Virus from a Chicken in Taiwan in 2008
Ming-Chu Cheng, Kosuke Soda, Ming-Shiuh Lee, Shu-Hwae Lee, Yoshihiro Sakoda, Hiroshi Kida, Ching-Ho Wang
AVIAN DISEASES, 54, 2, 885, 893, AMER ASSOC AVIAN PATHOLOGISTS, 2010年06月, ［査読有り］
英語, 研究論文（学術雑誌）, During the surveillance of avian influenza, an H5N2 influenza A virus was isolated from a cloacal swab sample of an apparently healthy chicken in Taiwan in October 2008. It was found that the HA of the virus had a pair of dibasic amino acid residues at the cleavage site, which might be a marker of highly pathogenic avian influenza virus. However, the intravenous pathogenicity index of the isolate was 0.89, indicating that the virus was approaching high pathogenicity in chickens. Virus isolation was negative in 2916 birds from 146 farms in a 3-km radius around the farm where the virus was isolated. Genetic analysis of the eight segments of the isolate indicated that the isolated virus was a reassortant whose HA and NA gene segments belonged to the American lineage and internal genes to the Eurasian lineage.

[Looking back upon the confusion of avian, swine, and pandemic influenza].
Kida H
Uirusu, 60, 1, 17, 20, 1, 2010年06月, ［査読有り］
日本語, "鳥インフルエンザ"，"ブタインフルエンザ"，そして"新型インフルエンザ"はいずれも，ヒトの病名ではない．すべて「インフルエンザ」である．日本だけで毎年数千名を死亡させ，数百名の小児に脳症を起こしている季節性インフルエンザの克服こそが，パンデミックインフルエンザ対策の基盤である．

Increase in the Neuraminidase Activity of a Nonpathogenic Newcastle Disease Virus Isolate during Passaging in Chickens
Ryota Tsunekuni, Hiroshi Ito, Hiroshi Kida, Koichi Otsuki, Toshihiro Ito
JOURNAL OF VETERINARY MEDICAL SCIENCE, 72, 4, 453, 457, JAPAN SOC VET SCI, 2010年04月, ［査読有り］
英語, 研究論文（学術雑誌）, A pathogenic mutant of the Newcastle disease virus (NDV) was previously generated by passaging a non-pathogenic isolate from wild waterfowl. Velogenic mutant 9a5b (IVPI=2.67) contains three amino acid substitutions (128H, 495K and 573stop) in the hemagglutinin-neuraminidase (FIN) protein, as compared with nonpathogenic waterfowl isolate 415/91 strain, and two of these (128H and 495K) were introduced after mesogenic 9a3b (IVPI=1.88). To investigate the role of the HN protein in NDV virulence, the Function of HN protein such as neuraminidase (NA), Hemadsorption (H Ad) and fusion promotion activities was examined by introducing the point mutations observed in passaged mutants into the HN gene cDNAs. In vitro functional assay using mutant protein expression demonstrated that the 128H substitution markedly increases NA activity and 573stop substitution increase NA and HAd activities. On the other hand, 495K substitution had little effect on any activities. These results indicate that a single amino acid substitution (128P to H) in the NDV HN protein affects the neuraminidase activity and is possibly correlated with the virulence.

Genetic comparisons between lentogenic Newcastle disease virus isolated from waterfowl and velogenic variants
Ryota Tsunekuni, Hiroshi Ito, Koichi Otsuki, Hiroshi Kida, Toshihiro Ito
VIRUS GENES, 40, 2, 252, 255, SPRINGER, 2010年04月, ［査読有り］
英語, 研究論文（学術雑誌）, Avirulent Newcastle disease viruses (NDV) harbored by waterfowl have the potential to become virulent after transmission to and circulation within chicken populations. In order to investigate how virulent viruses are selected from an avirulent background, we compared the complete sequences of the avirulent NDV isolate Goose/Alaska/415/91 and its virulent variant strain 9a5b, which was obtained by nine and five passages in the chick air sac and brain, respectively. Seven amino acid substitutions were detected in the M, F, and HN proteins. Two were detected between variants 9a3b and 9a5b (128P to H and 495E to K in HN protein) that were passed through the brain. Pathogenicity determined by the MDT and IVPI tests also differed between 9a3b and 9a5b. These results suggest that in addition to the F cleavage site sequence, these two amino acids in HN protein are also related to the pathogenicity of NDV in chickens.

Lipopolysaccharide treatment and inoculation of influenza A virus results in influenza virus-associated encephalopathy-like changes in neonatal mice
Tomohisa Tanaka, Yuji Sunden, Yoshihiro Sakoda, Hiroshi Kida, Kenji Ochiai, Takashi Umemura
JOURNAL OF NEUROVIROLOGY, 16, 2, 125, 132, SPRINGER, 2010年04月, ［査読有り］
英語, 研究論文（学術雑誌）, Influenza virus-associated encephalopathy (IAE) is a highly mortal neural complication of influenza A virus (IAV) infection, mostly affecting children younger than 5 years old, and the brain pathology of IAE is characterized by peracute brain edema with evidence of an impaired blood-brain barrier. The pathogenesis of IAE is unknown, but hypercytokinemia of tumor necrosis factor (TNF)-alpha, interleukin (IL)-1 beta, and IL-6 is suspected of playing a central role in the development of IAE. Because the brain pathology of IAE is similar to that of septic encephalopathy due to endotoxemia, the effect of combined treatment of IAV and lipopolysaccharide (LPS) was tested using suckling mice. The results show that pulmonary infection with non-neurotropic IAV enhanced the neuropathogenicity of LPS and induced encephalopathy that was similar to IAE with respect to the occurrence of central nervous system (CNS) histopathology and the absence of direct infection of IAV in the brain. Influenza A virus also increased blood-brain barrier (BBB) permeability and induced inflammatory cytokines in the blood. These results suggested that the mice treated with IAV+LPS are possible animal models of IAE, and that hypercytokinemia and/or the involvement of endotoxemia in IAV infection are possible causes of IAE.

Amelioration of pneumonia with Streptococcus pneumoniae infection by inoculation with a vaccine against highly pathogenic avian influenza virus in a non-human primate mixed infection model
Taichiro Miyake, Kosuke Soda, Yasushi Itoh, Yoshihiro Sakoda, Hirohito Ishigaki, Tomoya Nagata, Hideaki Ishida, Misako Nakayama, Hiroichi Ozaki, Hideaki Tsuchiya, Ryuzo Torii, Hiroshi Kida, Kazumasa Ogasawara
JOURNAL OF MEDICAL PRIMATOLOGY, 39, 1, 58, 70, WILEY-BLACKWELL PUBLISHING, INC, 2010年02月, ［査読有り］
英語, 研究論文（学術雑誌）, Background
Highly pathogenic avian influenza virus (HPAIV) infection has a high mortality rate in humans. Secondary bacterial pneumonia with HPAIV infection has not been reported in human patients, whereas seasonal influenza viruses sometimes enhance bacterial pneumonia, resulting in substantial morbidity and mortality. Therefore, if HPAIV infection were accompanied by bacterial infection, an increase in mortality would be expected. We examined whether a vaccine against HPAIV prevents severe morbidity caused by mixed infection with HPAIV and bacteria.
Methods
H7N7 subtype of HPAIV and Streptococcus pneumoniae were inoculated into cynomolgus macaques with or without vaccination of inactivated whole virus particles.
Results
Vaccination against H7N7 HPAIV decreased morbidity caused by HPAIV and pneumonia caused by S. pneumoniae. Bacterial replication in lungs was decreased by vaccination against HPAIV, although the reduction in bacterial colonies was not significant.
Conclusions
Vaccination against HPAIV reduces pneumonia caused by bacterial superinfection and may improve prognosis of HPAIV-infected patients.

Subcutaneous inoculation of a whole virus particle vaccine prepared from a non-pathogenic virus library induces protective immunity against H7N7 highly pathogenic avian influenza virus in cynomolgus macaques
Yasushi Itoh, Hiroichi Ozaki, Hirohito Ishigaki, Yoshihiro Sakoda, Tomoya Nagata, Kosuke Soda, Norikazu Isoda, Taichiro Miyake, Hideaki Ishida, Kiyoko Okamoto, Misako Nakayama, Hideaki Tsuchiya, Ryuzo Torii, Hiroshi Kida, Kazumasa Ogasawara
VACCINE, 28, 3, 780, 789, ELSEVIER SCI LTD, 2010年01月, ［査読有り］
英語, 研究論文（学術雑誌）, Development of H7N7 highly pathogenic avian influenza virus (HPAIV) vaccines is an urgent issue since human cases of infection with this subtype virus have been reported and most humans have no immunity against H7N7 viruses. We made an H7N7 vaccine combining components from an influenza virus library of non-pathogenic type A influenza viruses Antibody and T cell recall responses specific against the vaccine strain were elicited by subcutaneous inoculation with the whole virus particle vaccine with or without alum as an adjuvant in cynomolgus macaques. No significant difference was observed in magnitude of antibody responses between vaccination with alum and vaccination without alum, though vaccination with alum induced longer recall responses of CD8(+) T cells than did vaccination without alum. After challenge with a subtype of H7N7 HPAIV, the virus was detected in nasal swabs of unvaccinated macaques for 8 days but only for I day in the animals vaccinated either with or without alum, although the macaques vaccinated with alum showed elevated body temperature more briefly after infection. These findings demonstrated that this H7N7 HPAIV strain is pathogenic to macaques and that the vaccine conferred protective immunity to macaques against H7N7 HPAIV infection. (C) 2009 Elsevier Ltd. All rights reserved.

Predicting the Antigenic Structure of the Pandemic (H1N1) 2009 Influenza Virus Hemagglutinin
Manabu Igarashi, Kimihito Ito, Reiko Yoshida, Daisuke Tomabechi, Hiroshi Kida, Ayato Takada
PLOS ONE, 5, 1, e8553, PUBLIC LIBRARY SCIENCE, 2010年01月, ［査読有り］
英語, 研究論文（学術雑誌）, The pandemic influenza virus (2009 H1N1) was recently introduced into the human population. The hemagglutinin (HA) gene of 2009 H1N1 is derived from "classical swine H1N1'' virus, which likely shares a common ancestor with the human H1N1 virus that caused the pandemic in 1918, whose descendant viruses are still circulating in the human population with highly altered antigenicity of HA. However, information on the structural basis to compare the HA antigenicity among 2009 H1N1, the 1918 pandemic, and seasonal human H1N1 viruses has been lacking. By homology modeling of the HA structure, here we show that HAs of 2009 H1N1 and the 1918 pandemic virus share a significant number of amino acid residues in known antigenic sites, suggesting the existence of common epitopes for neutralizing antibodies cross-reactive to both HAs. It was noted that the early human H1N1 viruses isolated in the 1930s-1940s still harbored some of the original epitopes that are also found in 2009 H1N1. Interestingly, while 2009 H1N1 HA lacks the multiple N-glycosylations that have been found to be associated with an antigenic change of the human H1N1 virus during the early epidemic of this virus, 2009 H1N1 HA still retains unique three-codon motifs, some of which became N-glycosylation sites via a single nucleotide mutation in the human H1N1 virus. We thus hypothesize that the 2009 H1N1 HA antigenic sites involving the conserved amino acids will soon be targeted by antibody-mediated selection pressure in humans. Indeed, amino acid substitutions predicted here are occurring in the recent 2009 H1N1 variants. The present study suggests that antibodies elicited by natural infection with the 1918 pandemic or its early descendant viruses play a role in specific immunity against 2009 H1N1, and provides an insight into future likely antigenic changes in the evolutionary process of 2009 H1N1 in the human population.

Intranasal administration of a live non-pathogenic avian H5N1 influenza virus from a virus library confers protective immunity against H5N1 highly pathogenic avian influenza virus infection in mice: Comparison of formulations and administration routes of vaccines
Yoshitaka Kashima, Mizuho Ikeda, Yasushi Itoh, Yoshihiro Sakoda, Tomoya Nagata, Taichiro Miyake, Kosuke Soda, Hiroichi Ozaki, Misako Nakayama, Hitomi Shibuya, Masatoshi Okamatsu, Hirohito Ishigaki, Hideaki Ishida, Toshihiro Sawai, Yoshihiro Kawaoka, Hiroshi Kida, Kazumasa Ogasawar
VACCINE, 27, 52, 7402, 7408, ELSEVIER SCI LTD, 2009年12月, ［査読有り］
英語, 研究論文（学術雑誌）, Outbreaks of highly pathogenic avian influenza viruses (HPAIVs) would cause disasters worldwide. Various strategies against HPAIVs are required to control damage. it is thought that the use of non-pathogenic avian influenza viruses as live vaccines will be effective in an emergency, even though there might be some adverse effects, because small amounts of live vaccines will confer immunity to protect against HPAIV infection. Therefore, live vaccines have the advantage of being able to be distributed worldwide soon after an outbreak. In the present study, we found that intranasal administration of a live H5N1 subtype non-pathogenic virus induced antibody and cytotoxic T lymphocyte responses and protected mice against H5N1 HPAIV infection. In addition, it was found that a small amount (100 PFU) of the live vaccine was as effective as 100 mu g (approximately 10(10-11) PFU of virus particles)of the inactivated whole particle vaccine in mice. Consequently, the use of live virus vaccines might be one strategy for preventing pandemics of HPAIVs in an emergency. (C) 2009 Elsevier Ltd. All rights reserved.

Effective Prevention against Rabies by Intracerebral Immunization in Mice
Jae-Ho Shin, Yoshihiro Sakoda, Shiori Yano, Kenji Ochiai, Hiroshi Kida, Takashi Umemura
JOURNAL OF VETERINARY MEDICAL SCIENCE, 71, 10, 1331, 1336, JAPAN SOC VET SCI, 2009年10月, ［査読有り］
英語, 研究論文（学術雑誌）, To evaluate the efficacy of intracerebral (IC) immunization, mice were immunized with a rabies vaccine by the subcutaneous (SC), intramuscular (IM) or IC route, and 10-fold the 50% lethal dose of rabies virus was inoculated into the hindleg of the immunized or non-immunized mice. The antibody titer in serum was elevated and boosted by additional immunization via all routes, but highest after the IC immunization followed by the IM and SC routes, in this order. Intracerebrally immunized mice were completely protected from death and the neurological signs of infection, whereas the IM or SC immunization only partly protected the mice. In mouse models, IC immunization is more effective at inducing a protective immune response against the transneural spread of rabies virus than IM or SC immunization.

Characterization of H3N6 avian influenza virus isolated from a wild white pelican in Zambia
Edgar Simulundu, Aaron S. Mweene, Daisuke Tomabechi, Bernard M. Hang'ombe, Akihiro Ishii, Yuka Suzuki, Ichiro Nakamura, Hirofumi Sawa, Chihiro Sugimoto, Kimihito Ito, Hiroshi Kida, Lewis Saiwana, Ayato Takada
ARCHIVES OF VIROLOGY, 154, 9, 1517, 1522, SPRINGER WIEN, 2009年09月, ［査読有り］
英語, 研究論文（学術雑誌）, We characterized an influenza virus isolated from a great white pelican in Zambia. Phylogenetic analysis showed that all of its gene segments belonged to the Eurasian lineage and that they appear to have evolved in distinct geographical regions in Europe, Asia, and Africa, suggesting reassortment of virus genes maintained in wild aquatic birds whose flyways overlap across these continents. It is notable that this virus might possess some genes of the same origin as those of highly pathogenic H7 and H5 viruses isolated in Eurasia. The present study underscores the need for continued monitoring of avian influenza viruses in Eurasia and Africa.

Long lasting immunity in chickens induced by a single shot of influenza vaccine prepared from inactivated non-pathogenic H5N1 virus particles against challenge with a highly pathogenic avian influenza virus
Takashi Sasaki, Norihide Kokumai, Toshiaki Ohgitani, Ryuichi Sakamoto, Noriyasu Takikawa, Zhifeng Lin, Masatoshi Okamatsu, Yoshihiro Sakoda, Hiroshi Kida
VACCINE, 27, 38, 5174, 5177, ELSEVIER SCI LTD, 2009年08月, ［査読有り］
英語, 研究論文（学術雑誌）, An influenza vaccine was prepared from inactivated whole particles of the non-pathogenic strain A/duck/Hokkaido/Vac-1/04 (H5N1) virus using an oil adjuvant containing anhydromannitol-octaclecenoate-ether (AMOE). The vaccine was injected intramuscularly into five 4-week-old chickens, and 138 weeks after vaccination, they were challenged intranasally with 100 times 50% chicken lethal dose of the highly pathogenic avian influenza (HPAI) virus A/chicken/Yamaguchi/7/04 (H5N1). All 5 chickens survived without exhibiting clinical signs of influenza, although 2 days post-challenge, 3 vaccinated chickens shed limited titres of viruses in laryngopharyngeal swabs. (C) 2009 Elsevier Ltd. All rights reserved.

In vitro and in vivo characterization of new swine-origin H1N1 influenza viruses
Yasushi Itoh, Kyoko Shinya, Maki Kiso, Tokiko Watanabe, Yoshihiro Sakoda, Masato Hatta, Yukiko Muramoto, Daisuke Tamura, Yuko Sakai-Tagawa, Takeshi Noda, Saori Sakabe, Masaki Imai, Yasuko Hatta, Shinji Watanabe, Chengjun Li, Shinya Yamada, Ken Fujii, Shin Murakami, Hirotaka Imai, Satoshi Kakugawa, Mutsumi Ito, Ryo Takano, Kiyoko Iwatsuki-Horimoto, Masayuki Shimojima, Taisuke Horimoto, Hideo Goto, Kei Takahashi, Akiko Makino, Hirohito Ishigaki, Misako Nakayama, Masatoshi Okamatsu, Kazuo Takahashi, David Warshauer, Peter A. Shult, Reiko Saito, Hiroshi Suzuki, Yousuke Furuta, Makoto Yamashita, Keiko Mitamura, Kunio Nakano, Morio Nakamura, Rebecca Brockman-Schneider, Hiroshi Mitamura, Masahiko Yamazaki, Norio Sugaya, M. Suresh, Makoto Ozawa, Gabriele Neumann, James Gern, Hiroshi Kida, Kazumasa Ogasawara, Yoshihiro Kawaoka
NATURE, 460, 7258, 1021, U110, NATURE PUBLISHING GROUP, 2009年08月, ［査読有り］
英語, 研究論文（学術雑誌）, Influenza A viruses cause recurrent outbreaks at local or global scale with potentially severe consequences for human health and the global economy. Recently, a new strain of influenza A virus was detected that causes disease in and transmits among humans, probably owing to little or no pre-existing immunity to the new strain. On 11 June 2009 the World Health Organization declared that the infections caused by the new strain had reached pandemic proportion. Characterized as an influenza A virus of the H1N1 subtype, the genomic segments of the new strain were most closely related to swine viruses(1). Most human infections with swine-origin H1N1 influenza viruses (S-OIVs) seem to be mild; however, a substantial number of hospitalized individuals do not have underlying health issues, attesting to the pathogenic potential of S-OIVs. To achieve a better assessment of the risk posed by the new virus, we characterized one of the first US S-OIV isolates, A/California/04/09 ( H1N1; hereafter referred to as CA04), as well as several other S-OIV isolates, in vitro and in vivo. In mice and ferrets, CA04 and other S- OIV isolates tested replicate more efficiently than a currently circulating human H1N1 virus. In addition, CA04 replicates efficiently in non-human primates, causes more severe pathological lesions in the lungs of infected mice, ferrets and non-human primates than a currently circulating human H1N1 virus, and transmits among ferrets. In specific-pathogen-free miniature pigs, CA04 replicates without clinical symptoms. The assessment of human sera from different age groups suggests that infection with human H1N1 viruses antigenically closely related to viruses circulating in 1918 confers neutralizing antibody activity to CA04. Finally, we show that CA04 is sensitive to approved and experimental antiviral drugs, suggesting that these compounds could function as a first line of defence against the recently declared S- OIV pandemic.

Generation of congenic mouse strains by introducing the virus-resistant genes, Mx1 and Oas1b, of feral mouse-derived inbred strain MSM/Ms into the common strain C57BL/6J
Kanako Moritoh, Hideto Yamauchi, Atsushi Asano, Kentaro Yoshii, Hiroaki Kariwa, Ikuo Takashima, Norikazu Isoda, Yoshihiro Sakoda, Hiroshi Kida, Nobuya Sasaki, Takashi Agui
JAPANESE JOURNAL OF VETERINARY RESEARCH, 57, 2, 89, 99, HOKKAIDO UNIV, 2009年08月, ［査読有り］
英語, 研究論文（学術雑誌）, Mx1 (Myxovirus resistance protein) and Oas1b (Oligoadenylate synthetase-1), induced by type 1 interferon (IFN), play a role in early antiviral innate immunity by inhibiting the replication of viruses. In mice, Mx1 and Oas1b confer resistance to the infection of orthomyxoviruses including influenza viruses and flaviviruses including West Nile viruses, respectively. Laboratory mice have been used to study the mechanisms of the pathogenesis of these virus infections; however, it is possible that they are not a suitable model system to study these viruses, since most of the inbred laboratory mouse strains lack both genes. It has been reported that feral mouse-derived inbred strains show resistance to the infection of these viruses due to the presence of intact both genes. In this study, we generated congenic strains in which the Mx or Oas locus of the MSM/Ms (MSAI) mouce was introduced to the most widely used mouse strain, C57BL/6J (136). B6.MSM-Mx mice showed resistance to the infection of influenza virus but not of West Nile virus. On the other hand, B6.MSM-Oas mice showed resistance to the infection of West Nile virus but not of influenza virus. Our results indicate that Mx1 and Oas1b show highly antiviral specificity in mice possessing the same genetic background. Therefore, these congenic mice are useful for not only infection study but also investigation of host defense mechanism to these viruses.

Tetraspanin CD9 Negatively Regulates Lipopolysaccharide-Induced Macrophage Activation and Lung Inflammation
Mayumi Suzuki, Isao Tachibana, Yoshito Takeda, Ping He, Seigo Minami, Takeo Iwasaki, Hiroshi Kida, Sho Goya, Takashi Kijima, Mitsuhiro Yoshida, Toru Kumagai, Tadashi Osaki, Ichiro Kawase
JOURNAL OF IMMUNOLOGY, 182, 10, 6485, 6493, AMER ASSOC IMMUNOLOGISTS, 2009年05月, ［査読有り］
英語, 研究論文（学術雑誌）, Tetraspanins facilitate the formation of multiple molecular complexes at specialized membrane microdomains and regulate cell activation and motility. In the present study, the role of tetraspanin CD9 in LPS-induced macrophage activation and lung inflammation was investigated in vitro and in vivo. When CD9 function was ablated with mAb treatment, small interfering RNA transfection, or gene knockout in RAW264.7 cells or bone marrow-derived macrophages, these macrophages produced larger amounts of TNF-alpha, matrix metalloproteinase-2, and -9 upon stimulation with LPS in vitro, when compared with control cells. Sucrose gradient analysis revealed that CD9 partly colocalized with the LPS-induced signaling mediator, CD14, at low-density light membrane fractions. In CD9 knockout macrophages, CD14 expression, CD14 and TLR4 localization into the lipid raft, and their complex formation were increased whereas I kappa B alpha expression was decreased when compared with wild-type cells, suggesting that CD9 prevents the formation of LPS receptor complex. Finally, deletion of CD9 in mice enhanced macrophage infiltration and TNF-alpha production in the lung after intranasal administration of LPS in vivo, when compared with wild-type mice. These results suggest that macrophage CD9 negatively regulates LPS response at lipid-enriched membrane microdomains. The Journal of Immunology, 2009, 182: 6485-6493.

Factors responsible for plaque formation of A/duck/Siberia/272/1998 (H13N6) influenza virus on MDCK cells
Yoshimi Tsuda, Norikazu Isoda, Yoshihiro Sakoda, Hiroshi Kida
VIRUS RESEARCH, 140, 1-2, 194, 198, ELSEVIER SCIENCE BV, 2009年03月, ［査読有り］
英語, 研究論文（学術雑誌）, Many influenza A viruses form plaques on Madin-Darby canine kidney (MDCK) cells in the presence of trypsin. A/duck/Siberia/272/1998 (H13N6) (Sib272), however, does not form plaque on MDCK cells. After three blind passages of the strain on MDCK cells, plaque-forming variant was obtained and designated as A/duck/Siberia/272PF/1998 (H13N6) (Sib272PF). Genetic and functional analyses of Sib272 and Sib272PF revealed that amino acid substitutions, F3L of the HA2 subunit and T379K of the PB1, were responsible for plaque formation of Sib272PF by enhancing fusion and polymerase activities, respectively. (c) 2009 Elsevier B.V. All rights reserved.

Cross-Protective Potential of a Novel Monoclonal Antibody Directed against Antigenic Site B of the Hemagglutinin of Influenza A Viruses
Reiko Yoshida, Manabu Igarashi, Hiroichi Ozaki, Noriko Kishida, Daisuke Tomabechi, Hiroshi Kida, Kimihito Ito, Ayato Takada
PLOS PATHOGENS, 5, 3, e1000350, PUBLIC LIBRARY SCIENCE, 2009年03月, ［査読有り］
英語, 研究論文（学術雑誌）, The hemagglutinin (HA) of influenza A viruses has been classified into sixteen distinct subtypes (H1-H16) to date. The HA subtypes of influenza A viruses are principally defined as serotypes determined by neutralization or hemagglutination inhibition tests using polyclonal antisera to the respective HA subtypes, which have little cross-reactivity to the other HA subtypes. Thus, it is generally believed that the neutralizing antibodies are not broadly cross-reactive among HA subtypes. In this study, we generated a novel monoclonal antibody (MAb) specific to HA, designated MAb S139/1, which showed heterosubtypic cross-reactive neutralization and hemagglutination inhibition of influenza A viruses. This MAb was found to have broad reactivity to many other viruses (H1, H2, H3, H5, H9, and H13 subtypes) in enzyme-linked immunosorbent assays. We further found that MAb S139/1 showed neutralization and hemagglutination-inhibition activities against particular strains of H1, H2, H3, and H13 subtypes of influenza A viruses. Mutant viruses that escaped neutralization by MAb S139/1 were selected from the A/Aichi/2/68 (H3N2), A/Adachi/2/57 (H2N2), and A/WSN/33 (H1N1) strains, and sequence analysis of the HA genes of these escape mutants revealed amino acid substitutions at positions 156, 158, and 193 (H3 numbering). A molecular modeling study showed that these amino acids were located on the globular head of the HA and formed a novel conformational epitope adjacent to the receptor-binding domain of HA. Furthermore, passive immunization of mice with MAb S139/1 provided heterosubtypic protection. These results demonstrate that MAb S139/1 binds to a common antigenic site shared among a variety of HA subtypes and neutralizes viral infectivity in vitro and in vivo by affecting viral attachment to cells. The present study supports the notion that cross-reactive antibodies play some roles in heterosubtypic immunity against influenza A virus infection, and underscores the potential therapeutic utility of cross-reactive antibodies against influenza.

PB2 Protein of a Highly Pathogenic Avian Influenza Virus Strain A/chicken/Yamaguchi/7/2004 (H5N1) Determines Its Replication Potential in Pigs
Rashid Manzoor, Yoshihiro Sakoda, Naoki Nomura, Yoshimi Tsuda, Hiroichi Ozaki, Masatoshi Okamatsu, Hiroshi Kida
JOURNAL OF VIROLOGY, 83, 4, 1572, 1578, AMER SOC MICROBIOLOGY, 2009年02月, ［査読有り］
英語, 研究論文（学術雑誌）, It has been shown that not all but most of the avian influenza viruses replicate in the upper respiratory tract of pigs (H. Kida et al., J. Gen. Virol. 75: 2183-2188, 1994). It was shown that A/chicken/Yamaguchi/7/2004 (H5N1) [Ck/Yamaguchi/04 (H5N1)] did not replicate in pigs (N. Isoda et al., Arch. Virol. 151: 1267-1279, 2006). In the present study, the genetic basis for this host range restriction was determined using reassortant viruses generated between Ck/Yamaguchi/04 (H5N1) and A/swine/Hokkaido/2/1981 (H1N1) [Sw/Hokkaido/81 (H1N1)]. Two in vivo-generated single-gene reassortant virus clones of the H5N1 subtype (virus clones 1 and 2), whose PB2 gene was of Sw/Hokkaido/81 (H1N1) origin and whose remaining seven genes were of Ck/Yamaguchi/04 (H5N1) origin, were recovered from the experimentally infected pigs. The replicative potential of virus clones 1 and 2 was further confirmed by using reassortant virus (rg-Ck-Sw/PB2) generated by reverse genetics. Interestingly, the PB2 gene of Ck/Yamaguchi/04 (H5N1) did not restrict the replication of Sw/Hokkaido/81 (H1N1), as determined by using reassortant virus rg-Sw-Ck/PB2. The rg-Sw-Ck/PB2 virus replicated to moderate levels and for a shorter duration than parental Sw/Hokkaido/81 (H1N1). Sequencing of two isolates recovered from the pigs inoculated with rg-Sw-Ck/PB2 revealed either the D256G or the E627K amino acid substitution in the PB2 proteins of the isolates. The D256G and E627K mutations enhanced viral polymerase activity in the mammalian cells, correlating with replication of virus in pigs. These results indicate that the PB2 protein restricts the growth of Ck/Yamaguchi/04 (H5N1) in pigs.

Evaluation of the potency, optimal antigen level and lasting immunity of inactivated avian influenza vaccine prepared from H5N1 virus
Takashi Sasaki, Norikazu Isoda, Kosuke Soda, Ryuichi Sakamoto, Kazue Saijo, Junko Hagiwara, Norihide Kokumai, Toshiaki Ohgitani, Takashi Imamura, Akira Sawata, Zhifeng Lin, Yoshihiro Sakoda, Hiroshi Kida
JAPANESE JOURNAL OF VETERINARY RESEARCH, 56, 4, 189, 198, HOKKAIDO UNIV, 2009年02月, ［査読有り］
英語, 研究論文（学術雑誌）, Test vaccines comprised of inactivated water-in-oil emulsions containing various antigen levels were prepared using a non-pathogenic H5N1 avian influenza (AI) virus, A/duck/Hokkaido/Vac-1/04 (H5N1). The potencies of these test vaccines were evaluated by two experiments. In the first experiment, the triangular relationship among the antigen levels of test vaccines, the hemagglutination inhibition (HI) antibody response, and the protective effect against challenge with a highly pathogenic avian influenza (HPAI) virus, A/chicken/Yamaguchi/7/04 (H5N1), was confirmed. Then lasting immunity of chickens after a single-shot vaccination was confirmed in the second experiment. As a result, complete protection after the challenge was observed in chickens immunized by test vaccines with an antigen level of 160 HA units/dose or higher. Thus, it was ascertained that the minimum antigen level in the AI vaccine was 160 RA units/dose, and the minimum HI antibody titer that could protect chickens from HPAI virus infection-related death was considered to be 1:16. Dose-dependent III antibody responses were observed in chickens after the vaccination. Thus, 640 RA units/dose was thought to be similar to the optimal antigen level. Alternatively, the HI antibody titers of chickens, injected with the vaccine containing 640 HA units/dose, were maintained at 1:181 or higher for 100 weeks after the single-shot vaccination.

Antigenic structure of the hemagglutinin of H9N2 influenza viruses
Masatoshi Okamatsu, Yoshihiro Sakoda, Noriko Kishida, Norikazu Isoda, Hiroshi Kida
ARCHIVES OF VIROLOGY, 153, 12, 2189, 2195, SPRINGER WIEN, 2008年12月, ［査読有り］
英語, 研究論文（学術雑誌）, The hemagglutinins (HAs) of H9 influenza viruses isolated from birds and mammals of different species were antigenically and genetically analyzed. Antigenic variants were selected from A/swine/Hong Kong/10/98 (H9N2) and A/duck/Hokkaido/13/00 (H9N2) in the presence of monoclonal antibodies (MAbs). Based on the reactivity patterns of these mutants with a panel of MAbs, at least five non-overlapping antigenic sites were defined using eight MAbs which recognized seven distinct epitopes on the H9 HA molecule. Based on the reactivity patterns with the panel of monoclonal antibodies, 21 H9N2 virus strains isolated from birds and mammals were divided into 7 antigenically distinct groups. The present findings indicate that it is important to monitor the antigenic variation in H9 influenza viruses. The panel of MAbs in the present study, thus, should be useful for detailed antigenic analysis of the H9 HAs for epidemiological studies, the selection of vaccine strains, and diagnosis.

Structural and Molecular Characterization of the Prefoldin beta Subunit from Thermococcus Strain KS-1
Hiroshi Kida, Yuri Sugano, Ryo Iizuka, Masahiro Fujihashi, Masafurni Yohda, Kunio Miki
JOURNAL OF MOLECULAR BIOLOGY, 383, 3, 465, 474, ACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD, 2008年11月, ［査読有り］
英語, 研究論文（学術雑誌）, Prefoldin (PFD) is a heterohexameric molecular chaperone that is found in eukaryotic cytosol and archaea. PFD is composed of alpha and beta subunits and forms a "jellyfish-like" structure. PFD binds and stabilizes nascent polypeptide chains and transfers them to group II chaperonins for completion of their folding. Recently, the whole genome of Thermococcus kodakaraensis KOD1 was reported and shown to contain the genes of two alpha and two beta subunits of PFD. The genome of Thermococcus strain KS-1 also possesses two sets of a (alpha 1 and alpha 2) and beta subunits (beta 1 and beta 2) of PFD (TsPFD). However, the functions and roles of each of these PFD subunits have not been investigated in detail. Here, we report the crystal structure of the TsPFD beta 1 subunit at 1.9 angstrom resolution and its functional analysis. TsPFD beta 1 subunits form a tetramer with four coiled-coil tentacles resembling the jellyfish-like structure of heterohexameric PFD. The beta hairpin linkers of beta 1 subunits assemble to form a beta barrel "body" around a central fourfold axis. Size-exclusion chromatography and multi-angle light-scattering analyses show that the beta 1 subunits form a tetramer at pH 8.0 and a dimer of tetramers at pH 6.8. The tetrameric beta 1 subunits can protect against aggregation of relatively small proteins, insulin or lysozyme. The structural and biochemical analyses imply that PFD beta 1 subunits act as molecular chaperones in living cells of some archaea. (C) 2008 Elsevier Ltd. All rights reserved.

Cross-Glade protective immunity of H5N1 influenza vaccines in a mouse model
Shin Murakami, Ayaka Iwasa, Kiyoko Iwatsuki-Horimoto, Mutsumi Ito, Maki Kiso, Hiroshi Kida, Ayato Takada, Chairul A. Nidom, Le Quynh Mai, Shinya Yamada, Hirotaka Imai, Yuko Sakai-Tagawa, Yoshihiro Kawaoka, Taisuke Horimoto
VACCINE, 26, 50, 6398, 6404, ELSEVIER SCI LTD, 2008年11月, ［査読有り］
英語, 研究論文（学術雑誌）, H5N1 highly pathogenic avian influenza viruses evolved into several clades, leading to appreciably distinct antigenicities of their hemagglutinins. As such, candidate H5N1 pre-pandemic vaccines for human use should be sought. Here, to evaluate fundamental immunogenic variations between H5N1 vaccines, we prepared four inactivated H5N1 test vaccines from different phylogenetic Glades (clade 1, 2.1, 2.2, and 2.3.4) in accordance with the WHO recommendation, and tested their cross-Glade immunity in a mouse model by vaccination followed by challenge with heterologous virulent viruses. All H5N1 vaccines tested provided full or partial cross-clade protective immunity, except one Glade 2.2-based vaccine, which did not protect mice from clade 2.3.4 virus challenge. Among the test vaccines, a clade 2.1-based vaccine possessed the broadest-spectrum cross-immunity. These results suggest that currently stockpiled pre-pandemic vaccines, especially clade 2.1-based vaccines, will likely be useful as backup vaccines in a pandemic situation, even one involving antigenic-drifted viruses. (c) 2008 Elsevier Ltd. All rights reserved.

Antigenic and genetic analysis of H5 influenza viruses isolated from water birds for the purpose of vaccine use
Kosuke Soda, Hiroichi Ozaki, Yoshihiro Sakoda, Norikazu Isoda, Yoshinari Haraguchi, Saori Sakabe, Noritaka Kuboki, Noriko Kishida, Ayato Takada, Hiroshi Kida
ARCHIVES OF VIROLOGY, 153, 11, 2041, 2048, SPRINGER WIEN, 2008年11月, ［査読有り］
英語, 研究論文（学術雑誌）, In order to prepare H5N1 influenza virus vaccine, the hemagglutinins (HAs) of 14 H5 virus isolates from water birds in Asia were antigenically and genetically analyzed. Phylogenetic analysis of the H5 HA genes revealed that 13 isolates belong to Eurasian and the other one to North American lineages. Each of the deduced amino acid sequences of the HAs indicated a non-pathogenic profile. Antigenic analysis using a panel of monoclonal antibodies recognizing six different epitopes on the HA of A/duck/Pennsylvania/10218/1984 (H5N2) and chicken antiserum to an H5N1 reassortant strain generated between A/duck/Mongolia/54/2001 (H5N2) and A/duck/Mongolia/47/2001 (H7N1), [R(Dk/Mong-Dk/Mong) (H5N1)] showed that the HAs of highly pathogenic avian influenza (HPAI) viruses currently circulating in Asia were antigenically closely related to those of the present isolates from water birds. Mice subcutaneously injected with formalin-inactivated R(Dk/Mong-Dk/Mong) were protected from challenge with 100 mouse lethal dose of A/Viet Nam/1194/2004 (H5N1). The present results support the notion that the H5 isolates and the reassortant H5N1 strain should be useful for vaccine preparation.

Phylogenic analysis of the M genes of influenza viruses isolated from free-flying water birds from their Northern Territory to Hokkaido, Japan
Rashid Manzoor, Yoshihiro Sakoda, Aaron Mweene, Yoshimi Tsuda, Noriko Kishida, Gui-Rong Bai, Ken-Ichiro Kameyama, Norikazu Isoda, Kosuke Soda, Michiko Naito, Hiroshi Kida
VIRUS GENES, 37, 2, 144, 152, SPRINGER, 2008年10月, ［査読有り］
英語, 研究論文（学術雑誌）, During 2000-2007, 218 influenza viruses of 28 different combinations of HA (H1-H13) and NA (N1-N9) subtypes were isolated from fecal samples of free-flying water birds at two distant lakes in Hokkaido, Japan. Phylogenic analysis of the matrix (M) genes of 67 strains, selected on the basis of their subtype combinations, revealed that A/duck/Hokkaido/W95/2006 (H10N8) was a reassortant whose M gene belonged to North American non-gull-avian and the other seven genes to Eurasian non-gull-avian lineages. The M genes of other 65 strains belonged to Eurasian non-gull-avian and the one to Eurasian-gull lineages. The M genes of 65 strains were grouped into three different sublineages, indicating that influenza viruses circulating in different populations of free-flying water birds have evolved independently in nature.

Hereditary motor and sensory neuropathy with proximal dominancy in the lower extremities, urinary disturbance, and paroxysmal dry cough
Shiroh Miura, Hiroki Shibata, Hiroshi Kida, Kazuhito Noda, Katsuro Tomiyasu, Ken Yamamoto, Akiko Iwaki, Mitsuyoshi Ayabe, Hisarnichi Aizawa, Takayuki Taniwaki, Yasuyuki Fukumaki
JOURNAL OF THE NEUROLOGICAL SCIENCES, 273, 1-2, 88, 92, ELSEVIER SCIENCE BV, 2008年10月, ［査読有り］
英語, 研究論文（学術雑誌）, We Studied a four-generation pedigree of a Japanese family with hereditary neuropathy to elucidate the genetic basis of this disease. Twelve members of the family were enrolled in this study. The clinical features were neurogenic muscle weakness with proximal dominancy in the lower extremities, sensory involvement, areflexia, fine postural tremors, painful Muscle cramps, elevated creatine kinase levels, recurrent paroxysmal dry cough, and neurogenic bladder.
We performed a genome-wide search using genetic loci spaced at about 13 Mb intervals. Although nine chromosomes (1, 3, 4, 5, 6, 10, 17, 19, and 22) had at least one region in which the logarithm of odds (LOD) Score was over 1.0, no loci fulfilled the Criteria for significant evidence of linkage. Moreover, we analyzed an extra 14 markers on 3p12-q13 (the locus of hereditary Motor and sensory neuropathy, proximal dominant form) and an extra five markers on 3p22-p24 (the locus of hereditary sensory neuropathy with chronic cough) and observed LOD scores of <-3 on both 3p12-q13 and 3p22-p24. Mutation scanning of the entire coding regions of the MPZ and PMP22 genes revealed no mutations. We conclude that the disorder described here is a newly classified hereditary motor and sensory neuropathy with autosomal dominant inheritance. (c) 2008 Elsevier B.V. All rights reserved.

Phylogenic analysis of the M genes of influenza viruses isolated from free-flying water birds from their Northern Territory to Hokkaido, Japan (Virus Genes DOI: 10.1007/s11262-008-0248-7)
Rashid Manzoor, Yoshihiro Sakoda, Aaron Mweene, Yoshimi Tsuda, Noriko Kishida, Gui-Rong Bai, Ken-Ichiro Kameyama, Norikazu Isoda, Kosuke Soda, Michiko Naito, Hiroshi Kida
Virus Genes, 37, 2, 153, 2008年10月, ［査読有り］
英語, 研究論文（学術雑誌）

Potency of an inactivated avian influenza vaccine prepared from a non-pathogenic H5N1 reassortant virus generated between isolates from migratory ducks in Asia
Norikazu Isoda, Yoshihiro Sakoda, Noriko Kishida, Kosuke Soda, Saori Sakabe, Ryuichi Sakamoto, Takashi Imamura, Masashi Sakaguchi, Takashi Sasaki, Norihide Kokumai, Toshiaki Ohgitani, Kazue Saijo, Akira Sawata, Junko Hagiwara, Zhifeng Lin, Hiroshi Kida
ARCHIVES OF VIROLOGY, 153, 9, 1685, 1692, SPRINGER WIEN, 2008年09月, ［査読有り］
英語, 研究論文（学術雑誌）, A reassortant influenza virus, A/duck/Hokkaido/Vac-1/2004 (H5N1) (Dk/Vac-1/04), was generated between non-pathogenic avian influenza viruses isolated from migratory ducks in Asia. Dk/Vac-1/04 (H5N1) virus particles propagated in embryonated chicken eggs were inactivated with formalin and adjuvanted with mineral oil to form a water-in-oil emulsion. The resulting vaccine was injected intramuscularly into chickens. The chickens were challenged with either of the highly pathogenic avian influenza virus strains A/chicken/Yamaguchi/7/2004 (H5N1) or A/swan/Mongolia/3/2005 (H5N1) at 21 days post-vaccination (p. v.), when the geometric mean serum HI titers of the birds was 64 with the challenge virus strains. The vaccinated chickens were protected from manifestation of disease signs upon challenge with either of the highly pathogenic avian influenza viruses. However, challenge virus was recovered at low titers from the birds at 2 and 4 days post-challenge (p.c.). All 3 chickens challenged at 6 days p.v. died, whereas 3 chickens challenged at 8 days p.v. survived. These results indicate that the present vaccine confers clinical protection and reduction of virus shedding against highly pathogenic avian influenza virus challenge and should be useful as an optional tool in emergency cases.

Genetic Analyses of an H3N8 Influenza Virus Isolate, Causative Strain of the Outbreak of Equine Influenza at the Kanazawa Racecourse in Japan in 2007
Mika Ito, Makoto Nagai, Yuji Hayakawa, Hirofumi Komae, Naruto Murakami, Syouichi Yotsuya, Shingo Asakura, Yoshihiro Sakoda, Hiroshi Kida
JOURNAL OF VETERINARY MEDICAL SCIENCE, 70, 9, 899, 906, JAPAN SOC VET SCI, 2008年09月, ［査読有り］
英語, 研究論文（学術雑誌）, In August 2007, an outbreak of equine influenza occurred among vaccinated racehorses with Japanese commercial equine influenza vaccine at Kanazawa Racecourse in Ishikawa prefecture in Japan. Apparent symptoms were pyrexia (38.2-41.0 degrees C) and nasal discharge with or without coughing, although approximately half of the infected horses were subclinical. All horses had been shot with a vaccine that contained two inactivated H3N8 influenza virus strains [A/equine/La Plata/93 (La Plata/93) of American lineage and A/ equine/Avesta/93 (Avesta/93) of European lineage] and an H7N7 strain (A/equine/Newmarket/1/77). Influenza virus, A/equine/ Kanazawa/1/2007 (H3N8) (Kanazawa/07), was isolated from one of the nasal swab samples of diseased horses. Phylogenetic analysis indicated that Kanazawa/07 was classified into the American sublineage Florida. In addition, four amino acid substitutions were found in the antigenic sites B and E in the HA1 subunit protein of Kanazawa/07 in comparison with that of La Plata/93. Hemagglutination-inhibition (HI) test using 16 serum samples from recovering horses revealed that 1.4- to 8-fold difference in titers between Kanazawa/ 07 and either of the vaccine strains. The present findings suggest that Japanese commercial inactivated vaccine contributed to reducing the morbidity rate and manifestation of the clinical signs of horses infected with Kanazawa/07 that may be antigenically different from the vaccine strains.

NS1-DIVAシステムを用いた国内製造鳥インフルエンザワクチン注射鶏のモニタリング　　　　　　　　　　　　　　　
竹山 夏実, 三成 健二, 坂元 隆一, 佐々木 崇, 瀧川 義康, 真瀬 昌司, 土屋 耕太郎, 岡松 正敏, 塚本 健司, 林 志鋒, 迫田 義博, 喜田 宏
日本獣医学会学術集会講演要旨集, 146回, 193, 193, (公社)日本獣医学会, 2008年09月
日本語

H2N5 influenza virus isolates from terns in Australia: genetic reassortants between those of the Eurasian and American lineages
Noriko Kishida, Yoshihiro Sakoda, Mai Shiromoto, Gui-Rong Bai, Norikazu Isoda, Ayato Takada, Graeme Laver, Hiroshi Kida
VIRUS GENES, 37, 1, 16, 21, SPRINGER, 2008年08月, ［査読有り］
英語, 研究論文（学術雑誌）, To investigate the prevalence of influenza viruses in feral water birds in the Southern Hemisphere, fecal samples of terns were collected on Heron Island, Australia, in December 2004. Six H2N5 influenza viruses were isolated. This is the first report of the isolation of the H2 subtype from shore birds in Australia. Phylogenetic analysis revealed that the M gene belonged to the American lineage of avian influenza viruses and the other genes belonged to the Eurasian lineages, indicating that genetic reassortment occurs between viruses of Eurasian and American lineages in free flying birds in nature.

Genetically destined potentials for N-linked glycosylation of influenza virus hemagglutinin
Manabu Igarashi, Kimihito Ito, Hiroshi Kida, Ayato Takada
VIROLOGY, 376, 2, 323, 329, ACADEMIC PRESS INC ELSEVIER SCIENCE, 2008年07月, ［査読有り］
英語, 研究論文（学術雑誌）, The addition of oligosaccharide side chains to influenza virus hemagglutinin (HA) is believed to facilitate viral escape from immune pressure in the human population. To determine the implicit potentials for acquisition of N-linked glycosylation, we analyzed the genetic background of 16 subtypes of avian influenza virus, some of which may be potential pandemic viruses in the future. We found a significant difference among HA subtypes in their genomic sequences to produce N-glycosylation sites. Notably, recently circulating avian influenza viruses of the H5 and H9 subtypes may have rather greater capacities to undergo mutations associated with glycosylation of HA than past pandemic viruses. We hypothesize that influenza viruses maintained in natural reservoirs could have different potentials for sustained circulation, depending on their HA subtypes, if introduced into the human population. (C) 2008 Elsevier Inc. All rights reserved.

Induction of cytotoxic T-lymphocyte and antibody responses against highly pathogenic avian influenza virus infection in mice by inoculation of apathogenic H5N1 influenza virus particles inactivated with formalin
Toshihiro Sawai, Yasushi Itoh, Hiroichi Ozaki, Norikazu Isoda, Kiyoko Okamoto, Yoshitaka Kashima, Yoshihiro Kawaoka, Yoshihiro Takeuchi, Hiroshi Kida, Kazumasa Ogasawara
IMMUNOLOGY, 124, 2, 155, 165, WILEY-BLACKWELL, 2008年06月, ［査読有り］
英語, 研究論文（学術雑誌）, We investigated whether a vaccine derived from an apathogenic reassortant type A H5N1 influenza strain could induce immune responses in vivo that mediated protection from highly pathogenic avian influenza virus infection in mice. After two subcutaneous immunizations with formalin-inactivated H5N1 whole virus particles (whole particle vaccine), significant killing specific for cells presenting a nucleoprotein peptide from the vaccine strain of the virus was observed. Similar vaccination with viruses treated with ether and formalin, which are commonly used for humans as ether-split vaccines, induced little or no cytotoxic T-cell response. Furthermore, whole particle vaccines of the apathogenic H5N1 strain were more effective than ether-split vaccines at inducing antibody production able to neutralize a highly pathogenic H5N1 strain. Finally, whole particle vaccines of H5N1 protected mice against infection by an H5N1 highly pathogenic avian influenza virus more effectively than did ether-split vaccines. These results suggest that formalin-inactivated virus particles of apathogenic strains are effective for induction of both cytotoxic T-lymphocyte and antibody responses against highly pathogenic avian influenza viruses in vivo, resulting in protection from infection by a highly pathogenic H5N1 virus.

Development of a pen-site test kit for the rapid diagnosis of H7 highly pathogenic avian influenza
Rashid Manzoor, Yoshihiro Sakoda, Saori Sakabe, Tsuyoshi Mochizuki, Yasuharu Namba, Yoshimi Tsuda, Hiroshi Kida
JOURNAL OF VETERINARY MEDICAL SCIENCE, 70, 6, 557, 562, JAPAN SOC VET SCI, 2008年06月, ［査読有り］
英語, 研究論文（学術雑誌）, As well as H5 highly pathogenic avian influenza viruses (HPAIV), H7 HPAIV strains have caused serious damages in poultry industries worldwide. Cases of bird-to-human transmission of H7 HPAIV have also been reported [I I]. On the outbreak of avian influenza, rapid diagnosis is critical not only for the control of HPAI but also for human health. In the present study, a rapid diagnosis kit based on immunochromatography for the detection of H7 hemagglutinin (HA) antigen of influenza A virus was developed using 2 monoclonal antibodies that recognize different epitopes on the H7 HAS. The kit detected each of the tested 15 H7 influenza virus strains and did not react with influenza A viruses of the other subtypes than H7 or other avian viral and bacterial pathogens. The kit detected H7 HA antigen in the swabs and tissue homogenates of the chickens experimentally infected with HPAIV strain A/chicken/Netherlands/2586/03 (H7N7). The results indicate that the present kit is specific and sensitive enough for the diagnosis of HPAI caused by H7 viruses, thus, recommended for the field application as a pen-site test kit.

Cleavage of the NS2-3 protein in the cells of cattle persistently infected with non-cytopathogenic bovine viral diarrhea virus
Ken-ichiro Kameyama, Yoshihiro Sakoda, Keita Matsuno, Asako Ito, Motoshi Tajima, Shigeyuki Nakamura, Hiroshi Kida
MICROBIOLOGY AND IMMUNOLOGY, 52, 5, 277, 282, BLACKWELL PUBLISHING, 2008年05月, ［査読有り］
英語, 研究論文（学術雑誌）, The NS2-3 of BVDV is cleaved in cultured cells infected with cp BVDV but not in those infected with ncp BVDV when tested more than 10 hours post infection. However, it is not known whether cleavage of NS2-3 occurs in vivo. In the present study, cleavage of NS2-3 in cattle persistently infected with BVDV was investigated. All BVDV isolated from PI animals were of the ncp biotype, and NS2-3 proteins were detected in bovine fetal muscular cells infected with these viruses. On the other hand, in the leukocytes of those PI animals, NS3 proteins, products of the cleavage of NS2-3 proteins, were detected. In addition, the NS3 proteins were also detected in leukocytes artificially infected with ncp BVDV. These results reveal that the NS2-3 protein of BVDV is cleaved in leukocytes. Furthermore, NS3 proteins were detected in many tissues of PI cattle, such as lymphoid tissue, brain, thyroid, lung, and kidney. These results suggest that the NS2-3 protein of ncp BVDV cleaves in vivo.

A vaccine prepared from a non-pathogenic H7N7 virus isolated from natural reservoir conferred protective immunity against the challenge with lethal dose of highly pathogenic avian influenza virus in chickens
Saori Sakabe, Yoshihiro Sakoda, Yoshinari Haraguchi, Norikazu Isoda, Kosuke Soda, Hiroki Takakuwa, Kazue Saijo, Akira Sawata, Katsumi Kume, Junko Hagiwara, Kotaro Tuchiya, Zhifeng Lin, Ryuichi Sakamoto, Takashi Imamura, Takashi Sasaki, Norihide Kokumai, Yoshihiro Kawaoka, Hiroshi Kida
VACCINE, 26, 17, 2127, 2134, ELSEVIER SCI LTD, 2008年04月, ［査読有り］
英語, 研究論文（学術雑誌）, During 2001-2004, 41 H7 influenza viruses (2 H7N1 and 39 H7N7 strains) were isolated from fecal samples of migratory ducks that flew from Siberia in the autumn of each year to Japan and Mongolia. A phylogenetic analysis of the hemagglutinin (HA) genes of the nine representative isolates revealed that they belonged to the Eurasian Lineage and the deduced amino acid sequence at the cleavage site of the HAs represented apathogenic profiles. One of the H7 isolates A/duck/Mongolia/736/02 (H7N7) was chosen from these H7 isolates for the preparation of the test vaccine. To improve the growth potential of A/duck/Mongolia/ 736/02 (H7N7) in chicken embryos, A/duck/Hokkaido/Vac-2/04 (H7N7) was generated by genetic reassortment between A/duck/Mongolia/736/02 (H7N7) as the donor of the PB2, PB1, PA, HA, NA, and NS genes and A/duck/Hokkaido/49/98 (H9N2) as that of NP and M genes. The test vaccine was prepared as follows; A/duck/Hokkaido/Vac-2/04 (H7N7) was propagated in chicken embryos and the virus in the allantoic fluid was inactivated and adjuvanted to form an oil-in-water emulsion. The test vaccine conferred immunity to chickens, completely protecting the manifestation of clinical signs against the challenge with lethal dose of H7 highly pathogenic avian influenza virus. These results indicate that influenza viruses isolated from natural reservoirs are useful for vaccine strains. (C) 2008 Elsevier Ltd. All rights reserved.

Identification of new genetic subtypes of bovine viral diarrhea virus genotype 1 isolated in Japan
Makoto Nagai, Michiko Hayashi, Mika Itou, Toyoko Fukutomi, Hiroomi Akashi, Hiroshi Kida, Yoshihiro Sakoda
VIRUS GENES, 36, 1, 135, 139, SPRINGER, 2008年02月, ［査読有り］
英語, 研究論文（学術雑誌）, A part of the nucleotide sequence of the 5' untranslated region (5'UTR) and E-rns region, and the genomic regions encoding for N-pro, Core, and E2 of So-like isolates and IS25CP/01 strain which belong to bovine viral diarrhea virus genotype 1 (BVDV-1) were determined and genetic comparisons were made with sequences of other BVDV subgenotypes. Phylogenetic analysis using the 5'UTR and N-pro revealed that So-like isolates and IS25CP/01 branched into independent phylogenetic branch. So-like isolates were clustered with Korean BVDV strains taken from DDBL/EMBL/GenBank in the 5'UTR. An additional two amino acid residues were found at the C terminal of the Core region of IS25CP/01. The similarity of amino acid sequence of E2 of So-like isolates and IS25CP/01 to the BVDV-1 reference strain NADL were 78.0-78.5 and 79.0, respectively. Cross-neutralization tests showed significant antigenic differences between So-like isolates and the others (Antigenic similarity (R) value: 2.2-8.8), and IS25CP/01 and the others (R value: 1.6-8.8). So-like viruses and IS25CP/01 differed from the thirteenth subgenotypes (1a-1m) reported by Jackova et al. (2007) and were classified as new genetic subtypes, BVDV-1n (So-like) and 1o (IS25CP/01).

Structure and molecular dynamics simulation of archaeal prefoldin: The molecular mechanism for binding and recognition of nonnative substrate proteins
Akashi Ohtaki, Hiroshi Kida, Yusuke Miyata, Naoki Ide, Akihiro Yonezawa, Takatoshi Arakawa, Ryo Iizuka, Keiichi Noguchi, Akiko Kita, Masafumi Odaka, Kunio Miki, Masafumi Yohda
JOURNAL OF MOLECULAR BIOLOGY, 376, 4, 1130, 1141, ACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD, 2008年02月, ［査読有り］
英語, 研究論文（学術雑誌）, Prefoldin (PFD) is a heterohexameric molecular chaperone complex in the eukaryotic cytosol and archaea with a jellyfish-like structure containing six long coiled-coil tentacles. PFDs capture protein folding intermediates or unfolded polypeptides and transfer them to group II chaperonins for facilitated folding. Although detailed studies on the mechanisms for interaction with unfolded proteins or cooperation with chaperonins of archaeal PFD have been performed, it is still unclear how PFD captures the unfolded protein. In this study, we determined the X-ray structure of Pyrococcus horikoshii OT3 PFD (PhPFD) at 3.0 angstrom resolution and examined the molecular mechanism for binding and recognition of nonnative substrate proteins by molecular dynamics (MD) simulation and mutation analyses. PhPFD has a jellyfish-like structure with six long coiled-coil tentacles and a large central cavity. Each subunit has a hydrophobic groove at the distal region where an unfolded substrate protein is bound. During MD simulation at 330 K, each coiled coil was highly flexible, enabling it to widen its central cavity and capture various normative proteins. Docking MD simulation of PhPFD with unfolded insulin showed that the beta subunit is essentially involved in substrate binding and that the alpha subunit modulates the shape and width of the central cavity. Analyses of mutant PhPFDs with amino acid replacement of the hydrophobic residues of the P subunit in the hydrophobic groove have shown that beta Ile107 has a critical role in forming the hydrophobic groove.
(C) 2007 Elsevier Ltd. All rights reserved.

A vaccine prepared from a non-pathogenic H5N1 avian influenza virus strain confers protective immunity against highly pathogenic avian influenza virus infection in cynomolgus macaques
Yasushi Itoh, Hiroichi Ozaki, Hideaki Tsuchiya, Kiyoko Okamoto, Ryuzo Torii, Yoshihiro Sakoda, Yoshihiro Kawaoka, Kazumasa Ogasawara, Hiroshi Kida
Vaccine, 26, 4, 562, 572, 4, 2008年01月24日, ［査読有り］
英語, 研究論文（学術雑誌）, In order to prepare for the emergence of pandemic influenza viruses, we have established an influenza virus library that contains non-pathogenic influenza A virus strains with 135 combinations of 15 hemagglutinin and 9 neuraminidase subtypes. In this study, we developed a vaccine against H5N1 highly pathogenic avian influenza (HPAI) virus infection in humans using a virus strain selected from the library. We examined its immunogenic potency using cynomolgus macaques as a primate model. Virus antigen-specific antibodies were elicited by intranasal or subcutaneous administration of inactivated whole virus particle vaccines. After challenge with an H5N1 HPAI virus isolate obtained from a Vietnamese patient, the virus was detected only on next day following inoculation in the nasal and/or tracheal swabs of vaccinated macaques that were asymptomatic. On the other hand, the viruses were isolated from nasal and tracheal swabs from non-vaccinated macaques until day 5 and day 7 after inoculation of the H5N1 HPAI virus, respectively. Although six non-vaccinated macaques developed a high body temperature, and two of them lost their appetite after HPAI virus infection, they recovered by the end of the 12-day observation period and did not show the severe symptoms that have been reported in human H5N1 virus infection cases. This demonstrates that the vaccine prepared with the non-pathogenic H5N1 virus from our influenza virus library conferred protective immunity against H5N1 HPAI virus infection to macaques. © 2007 Elsevier Ltd. All rights reserved.

Development of vaccine strains of H5 and H7 influenza viruses
Kosuke Soda, Yoshihiro Sakoda, Norikazu Isoda, Masahiro Kajihara, Yoshinari Haraguchi, Hitomi Shibuya, Hiromi Yoshida, Takashi Sasaki, Ryuichi Sakamoto, Kazue Saijo, Junko Hagiwara, Hiroshi Kida
JAPANESE JOURNAL OF VETERINARY RESEARCH, 55, 2-3, 93, 98, HOKKAIDO UNIV, 2008年01月, ［査読有り］
英語, 研究論文（学術雑誌）, To establish vaccine strains of H5 and H7 influenza viruses, A/duck/Hokkaido/Vac-1/04 (H5N1) [Vac-1/04 (H5N1)], A/duck/Hokkaido/Vac-3/07 (H5N1) [Vac-3/07 (H5N1)], and A/duck/Hokkaido/ Vac-2/04 (H7N7) [Vac-2/04 (H7N7)] were generated from non-pathogenic avian influenza viruses isolated from migratory ducks. Vac-1/04 (H5N1) and Vac-3/07 (H5N1) were generated by genetic reassortment between H5N2 or H5N3 virus as an HA gene provider and H7N1 or H6N1 viruses as an NA gene provider. Vac-2/04 (H7N7) was a genetic reassortant obtained using H7N7 and H9 N2 viruses to give high growth character of the H9N2 virus in chicken embryonated eggs. The results of sequence analyses and experimental infections revealed that these H5N1 and H7N7 reassortant viruses were non-pathogenic in chickens and embryos, and had good growth potential in embryonated eggs. These viruses should be useful to develop vaccines against H5 and H7 highly pathogenic avian influenza viruses.

Carboxyl terminus of the 34 kDa protein of Mycobacterium paratuberculosis shares homologous B-cell epitopes with Mycobacterium avium and Mycobacterium intracellulare.
Malamo M, Okazaki K, Sakoda Y, Kida H
The Veterinary record, 161, 853, 857, 25, 2007年12月, ［査読有り］

Effect of disinfactnts on infectivity of avian influenza viruses.　　　　　　　　　　　　　　　
Ito T, Kida H, Otsuki K
J. Anim. Hyg., 33, 61, 64, 2007年12月, ［査読有り］
英語, 研究論文（学術雑誌）

The quail and chicken intestine have sialyl-galactose sugar chains responsible for the binding of influenza A viruses to human type receptors.
Chao-Tan Guo, Noriko Takahashi, Hirokazu Yagi, Koichi Kato, Tadanobu Takahashi, Shuang-Qin Yi, Yong Chen, Toshihiro Ito, Koichi Otsuki, Hiroshi Kida, Yoshihiro Kawaoka, Kazuya I-P Jwa Hidari, Daisei Miyamoto, Takashi Suzuki, Yasuo Suzuki
Glycobiology, 17, 7, 713, 24, 7, 2007年07月, ［査読有り］, ［国際誌］
英語, The receptor specificity of influenza viruses is one factor that allows avian influenza viruses to cross the species barrier. The recent transmissions of avian H5N1 and H9N2 influenza viruses from chickens and/or quails to humans indicate that avian influenza viruses can directly infect humans without an intermediate host, such as pigs. In this study, we used two strains of influenza A virus (A/PR/8/34, which preferentially binds to an avian-type receptor, and A/Memphis/1/71, which preferentially binds to a human-type receptor) to probe the receptor specificities in host cells. Epithelial cells of both quail and chicken intestines (colons) could bind both avian- and human-type viruses. Infected cultured quail colon cells expressed viral protein and allowed replication of the virus strain A/PR/8/34 or A/Memphis/1/71. To understand the molecular basis of these phenomena, we further investigated the abundance of sialic acid (Sia) linked to galactose (Gal) by the alpha2-3 linkage (Siaalpha2-3Gal) and Siaalpha2-6Gal in host cells. In glycoprotein and glycolipid fractions from quail and chicken colon epithelial cells, there were some bound components of Sia-Gal linkage-specific lectins, Maackia amurensis agglutinin (specific for Siaalpha2-3 Gal) and Sambucus nigra agglutinin (specific for Siaalpha2-6Gal), indicating that both Siaalpha2-3Gal and Siaalpha2-6Gal exist in quail and chicken colon cells. Furthermore, we demonstrated by fluorescence high-performance liquid chromatography (HPLC) analysis that 5-N-acetylneuraminic acid was the main molecular species of Sia, and we demonstrated by multi-dimensional HPLC mapping and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry analysis that bi-antennary complex-type glycans alpha2-6 sialylated at the terminal Gal residue(s) are major (more than 79%) sialyl N-glycans expressed by intestinal epithelial tissues in both the chicken and quail. Taken together, these results indicate that quails and chickens have molecular characterization as potential intermediate hosts for avian influenza virus transmission to humans and could generate new influenza viruses with pandemic potential.

Genetic and pathobiological characterization of bovine viral diarrhea viruses recently isolated from cattle in Japan
Keita Matsuno, Yoshihiro Sakoda, Ken-ichiro Kameyama, Kyuzo Tamai, Asako Ito, Hiroshi Kida
JOURNAL OF VETERINARY MEDICAL SCIENCE, 69, 5, 515, 520, JAPAN SOC VET SCI, 2007年05月, ［査読有り］
英語, 研究論文（学術雑誌）, The 475 strains of bovine viral diarrhea virus (BVDV) isolated from cattle in 12 prefectures of Japan in the last 7 years were phylo.genetically classified as BVDV-1 or BVDV-2 on the basis of the nucleotide sequence of the 5'-untransiated region. BVDV-1 strains were further subtyped as 1a (101 strains), 1b (163), 1c (128), 1j (3), and So CP/75-like (1), and all of the 79 BVDV-2 strains belonged to subtype 2a. These 2a BVDVs contain two isolates that had high nucleotide identities with those of highly pathogenic BVDV-2 strains reported in North America (Pellerin et al., 1994). However, acute infection with severe mortality like North American outbreak was not observed and most of the present BVDV-2 strains were isolated from persistently infected (PI) cattle showing mild or no clinical sign. Moreover, it was revealed that 61.5% of the 39 PI cattle with cytopathogenic BVDVs did not show typical mucosal disease and 54.6% of the 405 PI animals only with non-cytopathogenic BVDVs were apparently healthy. The present results indicate that the prevention of the infection with an appropriate vaccine and active surveillance covering healthy cattle are required for the control of BVD.

Changes in Cellular Composition and Induction of Cell Death in the Spleen Are Correlated to the Disease Symptoms in Response to Influenza A Virus
Sato Kayoko, Fujikura Daisuke, Takada Ayato, Kida Hiroshi, Miyazaki Tadaaki
JOURNAL OF IMMUNOLOGY, 178, 2007年04月01日, ［査読有り］

Serological evidence of influenza A virus infection in Kuril harbor seals (Phoca vitulina stejnegeri) of Hokkaido, Japan
Kei Fujii, Chiharu Kakumoto, Mari Kobayashi, Sachiko Saito, Tatsuya Kariya, Yukiko Watanabe, Yoshihiro Sakoda, Hiroshi Kida, Masatsugu Suzuki
JOURNAL OF VETERINARY MEDICAL SCIENCE, 69, 3, 259, 263, JAPAN SOC VET SCI, 2007年03月, ［査読有り］
英語, 研究論文（学術雑誌）, For proper management and conservation of the Kuril harbor seal (Phoca vitulina stejnegeri) through disease control, serological analysis was performed for influenza A virus infection in free-ranging seals in Hokkaido, Japan. Serum samples were collected from seals at Nosappu (231 seals), Akkeshi (16) and Erimo (75), between 1998 and 2005, and were analyzed by ELISA. Antibodies to the influenza A virus were detected only in seals from Nosappu. The incidences were 11% (1/9), 3% (2/66), 12% (7/59) and 6% (5/77) in 1998, 2003, 2004 and 2005, respectively. These suggest sporadic infection. Because anti body-positive seals included juvenile seals in each year, the infections were considered to have been circulated since no later than the late 1990s until recent years. ELISA-positive sera were analyzed by hemagglutination inhibition (HI) tests to determine the subtypes. Antibodies to the H3 and H6 subtypes were detected in 10 and 2 sera, respectively. Two of the sera that had antibodies to the H6 subtype also had antibodies to the H3 subtype. These two seals were considered to have been infected with both the H3 and H6 subtypes. This is the first investigation to find antibodies to the H6 subtype in seals. Although the H6 subtype had been isolated only from avians, genetic analysis had suggested that the H6 subtype could become a novel mammalian pathogen. For definitive diagnosis, detection of the virus from the tissue or mucus of seals is required.

Detection of influenza virus hemagglutinin with randomly immobilized anti-hemagglutinin antibody on a carbon nanotube sensor
Seiji Takeda, Hiroichi Ozaki, Satoshi Hattori, Atsushi Ishii, Hiroshi Kida, Koichi Mukasa
JOURNAL OF NANOSCIENCE AND NANOTECHNOLOGY, 7, 3, 752, 756, AMER SCIENTIFIC PUBLISHERS, 2007年03月, ［査読有り］
英語, 研究論文（学術雑誌）, Carbon nanotube sensors were capable of detecting hemagglutinin binding to anti-hemagglutinin antibody immobilized on the sensor. The carbon nanotube sensors were fabricated by chemical vapor deposition method and it showed field effect transistor property. Anti-hemagglutinin antibody was immobilized by cross-linking on the reverse surface of the carbon nanotube sensor. The current between the source and the drain was measured after incubation of various concentration of hemagglutinin antigen with immobilized anti-hemagglutinin antibody. I-V-gate curve was obtained by plotting the current as a function of the potential applied to the back gate. The I-V-gate curve showed a positive shift in a manner dependent on the hemagglutinin antigen concentration after immobilization of anti-hemagglutinin while no shift was observed without immobilization of anti-hemagglutinin antibody on the surface. The sensitivity of the CNT sensor was higher than that of the QCM method even without controlling the orientation of the anti-hemagglutinin antibody This method constitutes a new tool to analyze interactions among biomolecules on a substrate.

H5N2鳥インフルエンザウイルスを接種したSFP鶏における抗体応答の長期観察と野外感染例の解析　　　　　　　　　　　　　　　
岡松 正敏, 加茂前 仁弥, 棚橋 美和, 迫田 義博, 真瀬 昌司, 塚本 健司, 喜田 宏, 山口 成夫
日本獣医学会学術集会講演要旨集, 143回, 195, 195, (公社)日本獣医学会, 2007年03月
日本語

Extensive accumulation of influenza virus NS1 protein in the nuclei causes effective viral growth in Vero cells
Hiroichi Ozaki, Hiroshi Kida
MICROBIOLOGY AND IMMUNOLOGY, 51, 5, 577, 580, CENTER ACADEMIC PUBL JAPAN, 2007年, ［査読有り］
英語, 研究論文（学術雑誌）, We previously showed that modified A/Puerto Rico/8/34 (PR8) influenza master strain had improved viral rescue and growth properties in African green monkey kidney (Vero) cell line by introducing NS gene of Vero-adapted A/England/1/53 (vaEng53). In the present study, it was found that the NS1 protein derived from vaEng53 was extensively accumulated in the nuclei than that of PR8. This accumulation was caused by 7 amino acid differences in C-terminal region of NS1 protein. These results suggest that specific accumulation of NS1 protein may contribute to efficient viral replication in Vero cells.

Serological evidence of leptospiral infection in pig populations in different districts in Japan
Michiko Naito, Yoshihiro Sakoda, Takayuki Kamikawa, Yoshiki Nitta, Kazuhiko Hirose, Mitsuaki Sakashita, Satoru Kurokawa, Hiroshi Kida
MICROBIOLOGY AND IMMUNOLOGY, 51, 6, 593, 599, CENTER ACADEMIC PUBL JAPAN, 2007年, ［査読有り］
英語, 研究論文（学術雑誌）, Serum samples were collected from 938 pigs of 24 farms in Hokkaido, Kagoshima, and Okinawa prefectures in Japan in 2001-2005. Enzyme-linked immunosorbent assay (ELISA) was used for the detection of antibodies to LipL32 antigen which is common to Leptospira interrogans. Samples positive in ELISA were then investigated by microscopic agglutination test for the identification of causal leptospires. Antibodies specific to leptospires of serovars Copenhageni, Bratislava, Australis and Javanica were detected in serum samples of pigs from each of the three districts. In addition, antibodies to leptospires of serovars Autummalis and Thrassovi were predominantly detected in those from Kagoshima. The present study, thus, revealed that leptospires belonging to different serovars prevail in the pig population in Japan. In addition, it is the first detection of antibodies to leptospires belonging to serovars Javanica and Thrassovi in pigs in Japan.

Development of an immunochromatographic kit for rapid diagnosis of h5 avian influenza virus infection
Yoshimi Tsuda, Yoshihiro Sakoda, Saori Sakabe, Tsuyoshi Mochizuki, Yasuharu Namba, Hiroshi Kida
MICROBIOLOGY AND IMMUNOLOGY, 51, 9, 903, 907, CENTER ACADEMIC PUBL JAPAN, 2007年, ［査読有り］
英語, 研究論文（学術雑誌）, Highly pathogenic avian influenza (HPAI) caused by the H5N1 subtype has given rise to serious damage in poultry industries in Asia. The virus has expanded its geographical range to Europe and Africa, posing a great risk to human health as well. For the control of avian influenza, a rapid diagnosis by detecting the causative virus and identifying its subtype is essential. In the present study, a rapid diagnosis kit combining immunochromatography with enzyme immunoassay which detects the H5 HA antigen of influenza A virus was developed using newly established anti-H5 HA monoclonal antibodies. The present kit specifically detected all of the H5 influenza viruses tested, and did not react with the other HA subtypes. H5 HA antigens were detected from swabs and tissue homogenates of chickens infected with HPAI virus strain A/chicken/Yamaguchi/7/04 (H5N1) from 2 days post inoculation. The kit showed enough sensitivity and specificity for the rapid diagnosis of HPAI.

Development of an immunochromatographic test kit for rapid detection of bovine viral diarrhea virus antigen.
Kameyama K, Sakoda Y, Tamai K, Igarashi H, Tajima M, Mochizuki T, Namba Y, Kida H
Journal of virological methods, 138, 1-2, 140, 146, 2006年12月, ［査読有り］

Development of ELISA to detect antibodies specific to Mycobacterium avium subsp paratuberculosis with truncated 34 kDa proteins
Mumeka Malamo, Yoshihiro Sakoda, Hiroichi Ozaki, Hiroshi Kida
JAPANESE JOURNAL OF VETERINARY RESEARCH, 54, 2-3, 99, 107, HOKKAIDO UNIV, 2006年11月, ［査読有り］
英語, 研究論文（学術雑誌）, To develop ELISA to detect antibodies specific to Mycobacterium avium subsp. paratuberculosis (M. paratuberculosis), the carboxyl termini of the 34 kDa proteins of M. paratuberculosis and Mycobacterium avium subsp. avium (M. avium) were expressed in Escherichia cola expression system. Antibodies specific to M. paratuberculosis were detected with the truncated 34 kDa protein of M. paratuberculosis in ELISA after pre-absorption of serum samples with the truncated 34 kDa protein of M. avium. All the serum samples from cattle confirmed to be infected with M. paratuberculosis were positive and those from healthy cattle were negative in the present ELISA system. These results indicate that the established ELISA detects antibodies specific to M. paratuberculosis with high specificity and sensitivity and is an useful tool for the screening of Johne's disease.

Seroepidemiological survey of morbillivirus infection in Kuril harbor seals (Phoca vitulina stejnegeri) of Hokkaido, Japan
Kei Fujii, Hiroki Sato, Chiharu Kakumoto, Mari Kobayashi, Sachiko Saito, Tatsuya Kariya, Yukiko Watanabe, Yoshihiro Sakoda, Chieko Kai, Hiroshi Kida, Masatsugu Suzuki
JAPANESE JOURNAL OF VETERINARY RESEARCH, 54, 2-3, 109, 117, HOKKAIDO UNIV, 2006年11月, ［査読有り］
英語, 研究論文（学術雑誌）, Serological analysis was performed to detect morbillivirus infection in Kuril harbor seals in Hokkaido, Japan. Serum samples were collected from the seals at Nosappu (231 sera), Akkeshi (16), and Erimo (75) between 1998 and 2005. Antibodies to phocine distemper virus (PDV) were detected by ELISA in seals from Nosappu and Erimo. Antibodies to PDV were found in 56% (5/9) of the sampled seals from Nosappu in 1998, versus only 5% (3/66) for 2003, 1% (1/79) for 2004, and 1% (1/77) for 2005. These suggest epidemic caused by the virus in or before 1998. As antibody-positive seals included juvenile seals in 2003 and 2005, sporadic infections of the virus are thought to have occurred in recent years. In Erimo, antibodies to PDV were found in 50% (14/28) of sampled seals in 2004, versus only 13% (1/8) for 1999, 7 (1/15) for 2003, and 0 % (0/24) for 2005. These suggest sporadic infection by the virus before 2003 and the epizootic between after autumn in 2003, when samples of 2003 were collected, and 2004. Since antibodies to canine distemper virus (CDV) were detected in one adult seal from Nosappu in each year from 2003 to 2005, sporadic infections of the virus were suggested. There were no difference in incidence of seals with. antibodies to the viruses between males and females and between juveniles and adults.

Assembly and budding of Ebolavirus
Takeshi Noda, Hideki Ebihara, Yukiko Muramoto, Ken Fujii, Ayato Takada, Hiroshi Sagara, Jin Hyun Kim, Hiroshi Kida, Heinz Feldmann, Yoshihiro Kawaoka
PLOS PATHOGENS, 2, 9, 864, 872, PUBLIC LIBRARY SCIENCE, 2006年09月, ［査読有り］
英語, 研究論文（学術雑誌）, Ebolavirus is responsible for highly lethal hemorrhagic fever. Like all viruses, it must reproduce its various components and assemble them in cells in order to reproduce infectious virions and perpetuate itself. To generate infectious Ebolavirus, a viral genome-protein complex called the nucleocapsid (NC) must be produced and transported to the cell surface, incorporated into virions, and then released from cells. To further our understanding of the Ebolavirus life cycle, we expressed the various viral proteins in mammalian cells and examined them ultrastructurally and biochemically. Expression of nucleoprotein alone led to the formation of helical tubes, which likely serve as a core for the NC. The matrix protein VP40 was found to be critical for transport of NCs to the cell surface and for the incorporation of NCs into virions, where interaction between nucleoprotein and the matrix protein VP40 is likely essential for these processes. Examination of virus-infected cells revealed that virions containing NCs mainly emerge horizontally from the cell surface, whereas empty virions mainly bud vertically, suggesting that horizontal budding is the major mode of Ebolavirus budding. These data form a foundation for the identification and development of potential antiviral agents to combat the devastating disease caused by this virus.

Pathogenicity of H5N1 influenza A viruses isolated in Vietnam between late 2003 and 2005
Yukiko Muramoto, Thi Quynh Mai Le, Lien Song Phuong, Tung Nguyen, Thu Ha Nguyen, Yuko Sakai-Tagawa, Taisuke Horimoto, Hiroshi Kida, Yoshihiro Kawaoka
JOURNAL OF VETERINARY MEDICAL SCIENCE, 68, 7, 735, 737, JAPAN SOC VET SCI, 2006年07月, ［査読有り］
英語, 研究論文（学術雑誌）, Since late 2003, highly pathogenic H5N1 influenza A viruses have spread among poultry and wild aquatic birds in Asian countries. Transmission of these viruses to humans can be lethal. Most human cases of infection with H5N1 viruses have occurred in Vietnam. Therefore, to understand the pathogenicity in mammals of these H5N1 viruses, we took viruses isolated from poultry (5 strains) and humans (2 strains) in Vietnam and tested their virulence in mice. The results showed that the H5N1 viruses from humans were pathogenic in mice and that one avian isolate was also pathogenic. These findings suggested that the H5N1 viruses circulating in poultry adapted during replication in humans or that strains pathogenic in mice were transmitted directly to humans.

Pathogenicity of a highly pathogenic avian influenza virus, A/chicken/Yamaguchi/7/04 (H5N1) in different species of birds and mammals
N Isoda, Y Sakoda, N Kishida, GR Bai, K Matsuda, T Umemura, H Kida
ARCHIVES OF VIROLOGY, 151, 7, 1267, 1279, SPRINGER WIEN, 2006年07月, ［査読有り］
英語, 研究論文（学術雑誌）, Outbreaks of highly pathogenic avian influenza (HPAI) have been occurring in domestic poultry in Asia since 1996. In the beginning of 2004, HPAI outbreaks were caused by H5N1 virus in two farms and a group of pet chickens in different areas of Japan. In the present study, the pathogenicity of A/chicken/Yamaguchi/7/04 (H5N1), which had been isolated from a dead chicken during the first outbreak in Japan, was assessed in chickens, quails, budgerigars, ducklings, mice, and miniature pigs by experimental infection. The virus was highly pathogenic to all the birds tested. Mice were susceptible to infection with a low mortality rate and miniature pigs were resistant to infection with the virus.

Rapid detection of avian influenza virus a and subtype H5N1 by single step multiplex reverse transcription-polymerase chain reaction
HL Wei, GR Bai, AS Mweene, YC Zhou, YL Cong, J Pu, SH Wang, H Kida, JH Liu
VIRUS GENES, 32, 3, 261, 267, SPRINGER, 2006年06月, ［査読有り］
英語, 研究論文（学術雑誌）, Outbreaks of H5N1 highly pathogenic avian influenza (HPAI) virus caused great economic losses to the poultry industry and resulted in human deaths in Thailand and Viet Nam in 2004. Rapid typing and subtyping of H5N1 viruses, especially from clinical specimens, are desirable for taking prompt control measures to prevent the spread of the disease. Here, we developed a set of oligonucleotide primers able to detect, type and subtype H5 and N1 influenza viruses in a single step multiplex reverse transcription-polymerase chain reaction (RT-PCR). RNA was extracted from allantoic fluid or from specimens with guanidinium isothiocyanate reagent. Reverse transcription and PCR were carried out with a mixture of primers specific for influenza viruses of type A, subtype H5 and N1 in a single reaction system under identical conditions. The amplified DNA fragments were analyzed by agarose gel electrophoresis. All the H5N1 viruses tested in the study and the experimental specimens presented three specific bands by the method established here. The results presented here suggest that the method described below is rapid and specific and, therefore, could be valuable in the rapid detection of H5N1 influenza viruses in clinics.

Molecular characterization of the hemagglutinin and neuraminidase genes of H5N1 influenza A viruses isolated from poultry in Vietnam from 2004 to 2005
Y Muramoto, TQM Le, LS Phuong, T Nguyen, TH Nguyen, Y Sakai-Tagawa, K Iwatsuki-Horimoto, T Horimoto, H Kida, Y Kawaoka
JOURNAL OF VETERINARY MEDICAL SCIENCE, 68, 5, 527, 531, JAPAN SOC VET SCI, 2006年05月, ［査読有り］
英語, 研究論文（学術雑誌）, Highly pathogenic H5N1 avian influenza A viruses have been spreading among domestic poultry, wild aquatic birds, and humans in many Asian countries since 2003. The largest number of patients, to date, infected with the H5N1 viruses are in Vietnam, where these viruses continue to cause outbreaks in domestic poultry. Here, we molecularly characterized the hemagglutinin and neuraminidase genes of nine H5N1 viruses isolated between January 2004 and August 2005 from domestic poultry in Vietnam. We found that several groups of highly pathogenic H5N1 avian influenza viruses are circulating among these birds, which suggests that H5N1 viruses of different lineages have been introduced into Vietnam multiple times.

Disease constraints for utilization of the African buffalo (Syncerus caffer) on game ranches in Zambia
Hetron M. Munang'andu, Victor M. Siamudaala, Andrew Nambota, John M. Bwalya, Musso Munyeme, Aaron S. Mweene, Ayato Takada, Hiroshi Kida
JAPANESE JOURNAL OF VETERINARY RESEARCH, 54, 1, 3, 13, HOKKAIDO UNIV, 2006年05月, ［査読有り］
英語, Eco-tourism depending on wildlife is becoming increasingly profitable and landowners are beginning to favor game farming and ecotourism. In these areas, large-scale translocation of wildlife involves a diversity of species and large populations. The African buffalo (Syncerus caffer) is one of the major tourist attractions in Zambia. It accounts for 8.7% and 12.4% of the total animal species hunted in the Game Management Areas and the total hunting revenue earned in Zambia, respectively. It is ecologically an important animal species essential for the purpose of habitat control and facilitating the provision of suitable grazing pastures. However, the rearing of the African buffalo on game ranches has been hampered by its carrier state of the Southern Africa Terroritory (SAT) serotypes of foot and mouth disease virus (FMD). The African buffalo is also known to be a carrier of Theileria parva lawrencei, the causative agent of corridor disease (CD) that continues to have devastating effects on the livestock industry in Zambia. In addition, the importation of buffaloes from countries with populations endemic to bovine tuberculosis is highly restricted. Veterinary regulations in Zambia, strongly advocate against the translocation of buffaloes from protected areas to private ranches for disease control purposes thereby mounting a considerable constraint on the economic and ecological viability of the industry. It is hoped that this review will motivate the relevant government authorities in exploiting ways in which this animal species play a central role in eco-tourism.

Ecology and epidemiology of anthrax in cattle and humans in Zambia
Victor M. Siamudaala, John M. Bwalya, Hetron M. Munang'andu, Peter G. Sinyangwe, Fred Banda, Aaron S. Mweene, Ayato Takada, Hiroshi Kida
JAPANESE JOURNAL OF VETERINARY RESEARCH, 54, 1, 15, 23, HOKKAIDO UNIV, 2006年05月, ［査読有り］
英語, Anthrax is endemic in Western and North-western Provinces of Zambia. The disease occurs throughout the year and impacts negatively on the economy of the livestock industry and public health in Zambia. During 1989-1995, there were 1, 626 suspected cases of anthrax in cattle in Western province and of these 51 were confirmed. There were 220 cases of human anthrax cases in 1990 alone and 248 cases during 1991-1998 with 19.1% and 7.7% case fatality rates, respectively. Interplay of the ecology of affected areas and anthropogenic factors seem to trigger anthrax epidemics. Anthrax has drawn considerable attention in recent years due to its potential use as a biological weapon. In this paper, the history, current status and approaches towards the control of the disease in Zambia are discussed. Quarantine measures restrict trade of livestock and exchange of animals for draught power resulting in poor food security at household levels. Challenges of anthrax control are complex and comprise of socio-political, economical, environmental and cultural factors. Inadequate funding, lack of innovative disease control strategies and lack of cooperation from stakeholders are the major constraints to the control of the disease. It is hoped that the information provided here will stimulate continued awareness for the veterinary and medical authorities to maintain their surveillance and capabilities against the disease. This may lead to a culminating positive impact on livestock and human health in the southern African region.

Hierarchy among viral RNA (vRNA) segments in their role in vRNA incorporation into influenza A virions
Y Muramoto, A Takada, K Fujii, T Noda, K Iwatsuki-Horimoto, S Watanabe, T Horimoto, H Kida, Y Kawaoka
JOURNAL OF VIROLOGY, 80, 5, 2318, 2325, AMER SOC MICROBIOLOGY, 2006年03月, ［査読有り］
英語, 研究論文（学術雑誌）, The genome of influenza A viruses comprises eight negative-strand RNA segments. Although all eight segments must be present in cells for efficient viral replication, the mechanism(s) by which these viral RNA (vRNA) segments are incorporated into virions is not fully understood. We recently found that sequences at both ends of the coding regions of the HA, NA, and NS vRNA segments of A/WSN/33 play important roles in the incorporation of these vRNAs into virions. In order to similarly identify the regions of the PB2, PB1, and PA vRNAs of this strain that are critical for their incorporation, we generated a series of mutant vRNAs that possessed the green fluorescent protein gene flanked by portions of the coding and noncoding regions of the respective segments. For all three polymerase segments, deletions at the ends of their coding regions decreased their virion incorporation efficiencies. More importantly, these regions not only affected the incorporation of the segment in which they reside, but were also important for the incorporation of other segments. This effect was most prominent with the PB2 vRNA. These findings suggest a hierarchy among vRNA segments for virion incorporation and may imply intersegment association of vRNAs during virus assembly.

Genetic recombination at different points in the N-pro-coding region of bovine viral diarrhea viruses and the potentials to change their antigenicities and pathogenicities
K Kameyama, Y Sakoda, K Tamai, M Nagai, H Akashi, H Kida
VIRUS RESEARCH, 116, 1-2, 78, 84, ELSEVIER SCIENCE BV, 2006年03月, ［査読有り］
英語, 研究論文（学術雑誌）, Cytopathogenic (cp) bovine viral diarrhea virus (BVDV) strain KS86-1cp was isolated from a cow persistently infected with non-cytopathogenic (ncp) BVDV strain KS86-1ncp after development of mucosal disease by superinfection with cp BVDV strain Nose. cp BVDV strains 799cp and 839cp were also isolated from independent cattle that developed mucosal disease by superinfection with cpBVDV KS86-1cp. In the present Study, genetic analysis revealed that the genes of cp BVDV strains 799cp and 839cp were chimeras between the genes of the persisting ncp BVDVs and that of super infecting KS86-1cp. The genetic recombination that generates 799cp occurred between the identical points in the N-PRO gene region. whereas genetic recombination that generates 839cp Occurred between different points in the N-PRO gene region. Both 799cp and 839cp were inherited Jiv gene of KS86-1cp strain and envelope protein genes of the persisting Viruses. In addition, neutralization test disclosed that antigenicities of 799cp, 839cp, and KS86-1cp were also similar to each persisting virus. These findings indicate that exogenous cp BVDV containing insertion of Jiv gene in the 5 terminal region can induce genetic recombination with the original ncp BVDV at different points in the N-PRO gene region, and those viruses have high potential to change those antigenicities and pathogenicities by RNA recombination. (c) 2005 Elsevier B.V. All rights reserved.

The amino-terminal residue of glycoprotein B is critical for neutralization of bovine herpesvirus 1
K Okazaki, S Fujii, A Takada, H Kida
VIRUS RESEARCH, 115, 2, 105, 111, ELSEVIER SCIENCE BV, 2006年02月, ［査読有り］
英語, 研究論文（学術雑誌）, In order to address the neutralization epitope on bovine herpesvirus 1 (BHV 1) glycoprotein B (gB), a panel of monoclonal antibodies (MAbs), a series of truncation forms of the glycoprotein and an MAb-escape mutant were used in this study. Immunocytochemistry on the truncations using MAbs against the glycoprotein revealed that the neutralization epitopes recognized by the MAbs lay between residues 1 and 52 of mature gB. Comparison of the sequences among the mutant, parent, and revertant viruses demonstrated that the amino-terminal residue of mature gB of the escape mutant was changed from Arg to Gln. These findings indicate that the amino-terminal residue of gB is critical for neutralization of BHV1. (c) 2005 Elsevier B.V. All rights reserved.

Highly pathogenic H5N1 influenza virus causes coagulopathy in chickens
Y Muramoto, H Ozaki, A Takada, CH Park, Y Sunden, T Umemura, Y Kawaoka, H Matsuda, H Kida
MICROBIOLOGY AND IMMUNOLOGY, 50, 1, 73, 81, CENTER ACADEMIC PUBL JAPAN, 2006年, ［査読有り］
英語, 研究論文（学術雑誌）, Severe hemorrhage at multiple organs is frequently observed in chickens infected with highly pathogenic avian influenza (HPAI) A viruses. In this study we examined whether HPAI virus infection leads to coagulation disorder in chickens. Pathological examinations showed that the fibrin thrombi were formed in arterioles at the lung, associated with the viral antigens in endothelial cells of chickens infected intravenously with HPAI virus. Hematological analyses of peripheral blood collected from the chickens revealed that coagulopathy was initiated at early stage of infection when viral antigens were detected only in the endothelial cells and monocytes/macrophages. Furthermore, gene expression of the tissue factor, the main initiator of blood coagulation, was upregulated in the spleen, lung, and brain of HPAI virus-infected chickens. These results suggest that dysfunction of endothelial cells and monocytes/macrophages upon HPAI virus infection may induce hemostasis abnormalities represented by the excessive blood coagulation and consumptive coagulopathy in chickens.

Architecture of ribonucleoprotein complexes in influenza A virus particles
T Noda, H Sagara, A Yen, A Takada, H Kida, RH Cheng, Y Kawaoka
NATURE, 439, 7075, 490, 492, NATURE PUBLISHING GROUP, 2006年01月, ［査読有り］
英語, 研究論文（学術雑誌）, In viruses, as in eukaryotes, elaborate mechanisms have evolved to protect the genome and to ensure its timely replication and reliable transmission to progeny. Influenza A viruses are enveloped, spherical or filamentous structures, ranging from 80 to 120 nm in diameter(1). Inside each envelope is a viral genome consisting of eight single-stranded negative- sense RNA segments of 890 to 2,341 nucleotides each(1). These segments are associated with nucleoprotein and three polymerase subunits, designated PA, PB1 and PB2; the resultant ribonucleoprotein complexes (RNPs) resemble a twisted rod ( 10 - 15 nm in width and 30 - 120 nm in length) that is folded back and coiled on itself(2-4). Late in viral infection, newly synthesized RNPs are transported from the nucleus to the plasma membrane, where they are incorporated into progeny virions capable of infecting other cells. Here we show, by transmission electron microscopy of serially sectioned virions, that the RNPs of influenza A virus are organized in a distinct pattern ( seven segments of different lengths surrounding a central segment). The individual RNPs are suspended from the interior of the viral envelope at the distal end of the budding virion and are oriented perpendicular to the budding tip. This finding argues against random incorporation of RNPs into virions(5), supporting instead a model in which each segment contains specific incorporation signals that enable the RNPs to be recruited and packaged as a complete set(6-12). A selective mechanism of RNP incorporation into virions and the unique organization of the eight RNP segments may be crucial to maintaining the integrity of the viral genome during repeated cycles of replication.

Improvement of a rapid diagnosis kit to detect either influenza a or B virus infections
GR Bai, Y Sakoda, AS Mweene, N Fujii, H Minakawa, H Kida
JOURNAL OF VETERINARY MEDICAL SCIENCE, 68, 1, 35, 40, JAPAN SOC VET SCI, 2006年01月, ［査読有り］
英語, 研究論文（学術雑誌）, To improve the sensitivity of a kit, ESPLINE (R) INFLUENZA A&B for rapid diagnosis of influenza by detecting influenza A or B virus specific nucleoproteins (NP), the ESPLINE (R) INFLUENZA A&B-N was developed by using newly established monoclonal antibodies (MAbs) to the respective NP molecule. MAbs FVA2-11 and FrB1-03 recognize the epitope on the amino acid region 59-130aa of the NP molecule of influenza A virus, and that on the region 72-191aa of the NP of influenza B virus, respectively. The new kit detected influenza A and B virus antigens with a detection limit of 10(2.0)-10(2.7) pfu/test, which is 4-1000 times higher than that of the original kit. Importantly, this kit detected each of influenza A viruses of the known hemagglutinin (HA) subtypes (H1-H15) including the H5N1 viruses recently isolated from human and avian sources in Asia. The kit also detected all of the 15 representative influenza B virus strains tested. The ESPLINE (R) INFLUENZA A&B-N is thus a rapid and highly sensitive and specific kit for the diagnosis of either influenza A or B virus infections.

Outbreak of highly pathogenic avian influenza in Japan and anti-influenza virus activity of povidone-iodine products
H Ito, T Ito, M Hikida, J Yashiro, A Otsuka, H Kida, K Otsuki
DERMATOLOGY, 212, 115, 118, KARGER, 2006年, ［査読有り］
英語, 研究論文（学術雑誌）, Objectives: On January 12, 2004, an outbreak of highly pathogenic avian influenza, caused by the H5N1 strain, occurred in a one-layer flock in Yamaguchi Prefecture, Japan. It had been 79 years since the last outbreak of avian influenza was confirmed in Japan. By February, 3 additional outbreaks had occurred (1 in Oita Prefecture and 2 in Kyoto Prefecture). Influenza viruses are enveloped viruses and are relatively sensitive to inactivation by lipid solvents, such as detergents. Infectivity is also rapidly destroyed by ether, sodium hypochlorite, povidone-iodine (PVP-I), peracetic acid and alcohol. However, these antiviral effects were only tested against human influenza A viruses. In the present study, the antiviral activity of PVP-I products against H5, H7 and H9 avian influenza A viruses, which had recently been transmitted to humans, were investigated. Methods:The in vitro antiviral activity of PVP-I products (2% PVP-I solution, 0.5% PVP-I scrub, 0.25% PVP-I palm, 0.23% PVP-I gargle, 0.23% PVP-I throat spray and 2% PVP-I solution for animals) against avian influenza A viruses [a highly pathogenic avian influenza virus, A/crow/Kyoto/T2/04 (H5N1; 10(6.5) EID50/0.1 ml), and 3 low pathogenic avian influenza A viruses, A/whistling swan/Shimane/499/838 (H5N3; 10(4.8) EID50/0.1 ml), A/whistling swan/Shimane/42/80 (H7N7; 10(5.5) EID50/0-1 ml) and A/duck/Hokkaido/26/99 (H9N2; 10(4.8) EID50/0-1 ml)] were investigated using embryonated hen's eggs. Results/Discussion: Viral infectious titers were reduced to levels below the detection limits by incubation for only 10 s with the PVP-I products used in this study. These results indicate that PVP-I products have virucidal activity against avian influenza A viruses. Therefore, the PVP-I products are useful in the prevention and control of human infection by avian influenza A viruses. Copyright (c) 2006 S. Karger AG, Basel.

Efficacy of intracerebral immunization against pseudorabies virus in mice
Jae-Ho Shin, Yoshihiro Sakoda, Jae Hoon Kim, Tomohisa Tanaka, Hiroshi Kida, Takashi Kimura, Kenji Ochiai, Takashi Umemura
MICROBIOLOGY AND IMMUNOLOGY, 50, 10, 823, 830, CENTER ACADEMIC PUBL JAPAN, 2006年, ［査読有り］
英語, 研究論文（学術雑誌）, To evaluate the efficacy of intracerebral (IQ immunization, mice were immunized with formalin-inactivated pseudorabies virus (PRV) by either subcutaneous (SQ or IC injection, and then 10(6) plaque-forming units of PRV were introduced into the hindleg of the immunized or non-immunized mice by intramuscular injection. The antibody titer in serum was elevated and boosted by additional immunization via both the SC and IC routes, but was higher after IC immunization. Intracerebrally immunized mice were completely protected from mortality and neurological signs, whereas all the non-immunized and 80% of the subcutaneously immunized mice died after developing neurological signs. In mouse models, IC immunization is more effective at inducing a protective immune response against the transneural spread of PRV than SC immunization.

Serologic investigation of influenza A virus infection in cetaceans from the western North Pacific and the Southern oceans
K Ohishi, T Maruyama, A Ninomiya, H Kida, R Zenitani, T Bando, Y Fujise, K Nakamatsu, N Miyazaki, AN Boltunov
MARINE MAMMAL SCIENCE, 22, 1, 214, 221, WILEY-BLACKWELL, 2006年01月, ［査読有り］
英語, 研究論文（学術雑誌）

Library of influenza virus strains for vaccine and diagnostic use against highly pathogenic avian influenza and human pandemics
H Kida, Y Sakoda
OIE/FAO International Scientific Conference on Avian Influenza, 124, 69, 72, KARGER, 2006年, ［査読有り］
英語, 研究論文（国際会議プロシーディングス）, To prepare for the emergence of pandemic influenza in birds and mammals including humans, we have carried out global surveillance of avian influenza. Influenza A viruses of 48 combinations of 15 HA and 9 NA subtypes out of 135 theoretical combinations have been isolated from faecal samples of ducks in Alaska, Siberia, Mongolia, Taiwan, China and Japan. So far, viruses of 73 other combinations have been generated by genetic reassortment in chicken embryos. Thus, avian influenza viruses of 121 combinations of HA and NA subtypes have been stocked for use in vaccine and diagnosis. Their pathogenicity, antigenicity, genetic information, and yield in chicken embryo have been analysed and registered in the database.

Efficacy of intracerebral immunization against pseudorabies virus in mice
Shin JH, Sakoda Y, Kim JH, Tanaka T, Kida H, Kimura T, Ochiai K, Umemura T
Microbiology and Immunology, 50, 10, 823, 830, 2006年, ［査読有り］
英語, 研究論文（学術雑誌）

Serological prevalence of Leptospira interrogans serovar Bratislava in horses in Mongolia
N Odontsetseg, D Boldbaatar, AS Mweene, H Kida
VETERINARY RECORD, 157, 17, 518, 519, BRITISH VETERINARY ASSOC, 2005年10月, ［査読有り］
英語, 研究論文（学術雑誌）

Effect of endoscopic transthoracic sympathicotomy on plasma natriuretic peptide concentrations in humans
Y Nakamura, H Yoshizawa, M Hirasawa, H Kida, Y Matsumoto, T Ueyama
CIRCULATION JOURNAL, 69, 9, 1079, 1083, JAPANESE CIRCULATION SOCIETY, 2005年09月, ［査読有り］
英語, 研究論文（学術雑誌）, Background The effects of endoscopic transthoracic sympathicotomy (ETS) on plasma natriuretic peptides concentrations in humans were examined in order to elucidate the role of the sympathetic nervous system in their regulation.
Methods and Results Thirty-seven patients with palmar hyperhidrosis underwent ETS. Cardiac functional indices were assessed by echocardiography, and plasma atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP) concentrations were measured before and after ETS. ETS caused decreases in heart rate, mean arterial pressure, systemic vascular resistance, and increases in left ventricular (LV) end-diastolic volume, stroke index, ejection fraction, and left atrial diameter. LV end-systolic volume and cardiac index remained unchanged. Following ETS, ANP increased from 10.7 +/- 5.9 to 24.7 +/- 16.8 pg/ml (p < 0.01), and BNP increased from 5.1 +/- 4.2 to 19.7 +/- 21.5 pg/ml (p < 0.01). From the multivariate regression analysis, ETS, age and gender were determined to be significant predictors of changes in the ANP and BNP concentrations. None of the hemodynamic parameters were useful as independent predictors.
Conclusion The plasma concentrations of ANP and BNP increased after ETS, independent of hemodynamic changes, and apparently because of the release of the inhibitory effects of the cardiac sympathetic nerves on natriuretic peptide secretion.

Application of carbon nanotubes for detecting anti-hemagglutinins based on antigen-antibody interaction
S Takeda, A Sbagyo, Y Sakoda, A Ishii, M Sawamura, K Sueoka, H Kida, K Mukasa, K Matsumoto
BIOSENSORS & BIOELECTRONICS, 21, 1, 201, 205, ELSEVIER ADVANCED TECHNOLOGY, 2005年07月, ［査読有り］
英語, 研究論文（学術雑誌）, Carbon nanotube sensors detected anti-hemagglutinin binding to immobilized hemagglutinins. An ultra-sensitive detection method for antibodies or antigens in serum is required. Hemagglutinins were immobilized on the reverse side of a carbon nanotube, thereby producing a source and a drain. Electrode pads covered each edge of the nanotube. The 1-V curves between the source and the drain were measured after incubation of anti-hemagglutinins with immobilized hemagglutinins in a buffered solution on the reverse side of the nanotube. The sensitivity of the CNT sensor was higher than that of an ELISA system. This method constitutes a new tool to analyze interaction among biomolecules on a substrate. (c) 2004 Elsevier B.V. All rights reserved.

Pathogenicity of H5 influenza viruses for ducks
N Kishida, Y Sakoda, N Isoda, K Matsuda, M Eto, Y Sunaga, T Umemura, H Kida
ARCHIVES OF VIROLOGY, 150, 7, 1383, 1392, SPRINGER WIEN, 2005年07月, ［査読有り］
英語, 研究論文（学術雑誌）, Four H5N1 highly pathogenic avian influenza (HPAI) viruses and an avirulent reassortant H5N1 virus were tested for their pathogenicity in domestic ducks. A/chicken/Yamaguchi/7/04 (H5N1) (Ck/Yamaguchi/04) isolated from a dead bird during the HPAI outbreak in Japan and A/duck/Yokohama/aq-10/03 (H5N1) (Dk/Yokohama/03) isolated from duck meat at a quarantine inspection for importation from China replicated in multiple organs including the brain of ducks. The ducks infected with Ck/Yamaguchi/04 did not show any clinical signs, while those infected with Dk/Yokohama/03 showed neurological signs. The ducks infected either with A/Hong Kong/483/97 (H5N1) orA/tern/SouthAfrica/61 (H5N3), or with an avirulent H5N1 reassortant, did not show any clinical signs. Virus-specific antibodies were detected in the sera of the ducks infected with each of the five strains tested, indicating that all of the viral strains infected and replicated in the birds. Dk/Yokohama/03 grew in multiple organs more rapidly than did Ck/Yamaguchi/04. Considerable titers of virus were detected in the brain of the ducks infected with Dk/Yokohama/03 and these birds showed neurological signs. The present results demonstrate that the pathogenicity of influenza viruses for ducks does not correlate with that for chickens and that replication of the virus in the brain is critical for ducks to show neurological signs.

Serological surveillance of canine leptospirosis in Mongolia
N Odontsetseg, Y Sakoda, H Kida
VETERINARY RECORD, 157, 4, 120, 121, BRITISH VETERINARY ASSOC, 2005年07月, ［査読有り］
英語, 研究論文（学術雑誌）

In vitro demonstration of neural transmission of avian influenza A virus
K Matsuda, T Shibata, Y Sakoda, H Kida, T Kimura, K Ochiai, T Umemura
JOURNAL OF GENERAL VIROLOGY, 86, 4, 1131, 1139, SOC GENERAL MICROBIOLOGY, 2005年04月, ［査読有り］
英語, 研究論文（学術雑誌）, Neural involvement following infections of influenza viruses can be serious. The neural transport of influenza viruses from the periphery to the central nervous system has been indicated by using mouse models. However, no direct evidence for neuronal infection has been obtained in vitro and the mechanisms of neural transmission of influenza viruses have not been reported. In this study, the transneural transmission of a neurotropic influenza A virus was examined using compartmentalized cultures of neurons from mouse dorsal root ganglia, and the results were compared with those obtained using the pseudorabies virus, a virus with well-established neurotransmission. Both viruses reached the cell bodies of the neurons via the axons. This is the first report on axonal transport of influenza A virus in vitro. In addition, the role of the cytoskeleton (microtubules, microfilaments and intermediate filaments) in the neural transmission of influenza virus was investigated by conducting cytoskeletal perturbation experiments. The results indicated that the transport of avian influenza A virus in the neurons was independent of microtubule integrity but was dependent on the integrity of intermediate filaments, whereas pseudorabies virus needed both for neural spread.

The vagus nerve is one route of transneural invasion for intranasally inoculated influenza A virus in mice　　　　　　　　　　　　　　　
Matsuda K, Shibata T, Sakoda Y, Kida H, Kimura T, Ochiai K, Umemura T
Journal of General Virology, 86, 1131, 1139, 2005年04月, ［査読有り］
英語, 研究論文（学術雑誌）

Nucleocapsid-like structures of Ebola virus reconstructed using electron tomography
T Noda, K Aoyama, H Sagara, H Kida, Y Kawaoka
JOURNAL OF VETERINARY MEDICAL SCIENCE, 67, 3, 325, 328, JAPAN SOC VET SCI, 2005年03月, ［査読有り］
英語, 研究論文（学術雑誌）, Electron tomography (ET) is a new technique for high resolution, three-dimensional (3D) reconstruction of pleiomorphic macromolecular complexes, such as virus components. By employing this technique, we resolved the 3D structure of Ebola virus nucleocapsid-like (NC-like) structures in the cytoplasm of cells expressing NP, VP24, and VP35: the minimum components required to form these NC-like structures. Reconstruction of these tubular NC-like structures of Ebola virus showed them to be composed of left-handed helices spaced at short intervals, which is structurally consistent with other non-segmented negative-strand RNA viruses.

Protective effect of IL-6 on alveolar epithelial cell death induced by hydrogen peroxide
H Kida, M Yoshida, S Hoshino, K Inoue, Y Yano, M Yanagita, T Kumagai, T Osaki, Tachibana, I, Y Saeki, Kawase, I
AMERICAN JOURNAL OF PHYSIOLOGY-LUNG CELLULAR AND MOLECULAR PHYSIOLOGY, 288, 2, L342, L349, AMER PHYSIOLOGICAL SOC, 2005年02月, ［査読有り］
英語, 研究論文（学術雑誌）, The goal of this study was to examine whether IL-6 could directly protect lung resident cells, especially alveolar epithelial cells, from reactive oxygen species (ROS)-induced cell death. ROS induced IL-6 gene expression in organotypic lung slices of wild-type (WT) mice. ROS also induced IL-6 gene expression in mouse primary lung fibroblasts, dose dependently. The organotypic lung slices of WT were more resistant to ROS-induced DNA fragmentation than those of IL-6-deficient (IL-6-/-) mice. WT resistance against ROS was abrogated by treatment with anti-IL-6 antibody. TdT-mediated dUTP nick end labeling stain and electron microscopy revealed that DNA fragmented cells in the IL-6-/- slice included alveolar epithelial cells and endothelial cells. In vitro studies demonstrated that IL-6 reduced ROS-induced A549 alveolar epithelial cell death. Together, these data suggest that IL-6 played an antioxidant role in the lung by protecting lung resident cells, especially alveolar epithelial cells, from ROS-induced cell death.

Viral and bacterial diseases in livestock in Mongolia
N Odontsetseg, AS Mweene, H Kida
JAPANESE JOURNAL OF VETERINARY RESEARCH, 52, 4, 151, 162, HOKKAIDO UNIV, 2005年02月, ［査読有り］
英語, This review focuses on the status of infectious diseases that are serious for animal health and have adverse economic effects in Mongolia. Data presented here are limited due to the lack of published or other easily available documents. Foot-and-mouth disease continues to cause substantial economic losses as exemplified by the outbreak of infection with serotype O PanAsia lineage virus. In the case of the 2001 outbreak, a 65% reduction in export revenues was recorded. In order to ascertain the free status of Mongolia from rinderpest, sero-epidemiological surveillance has been carried out since 2001.
In 2004, Mongolia was certified free from rinderpest by Office International des Epizooties (OIE). A sharp rise in both animal and human brucellosis incidence has become a serious problem. Rabies and anthrax remain endemic with occasional human cases. Other prevailing infectious diseases are contagious pustular dermatitis, contagious agalactia, enterotoxemia and pasteurellosis. The current programs for the control of infectious diseases in livestock in Mongolia lack a definite policy that would enable rapid implementation. A large-scale surveillance of infectious diseases in animals and management of appropriate preventive measures are urgently required in Mongolia.

Erratum: Serological evidence of the persistence of infection with Leptospira interrogans serovar hardjo in cattle in Mongolia (Microbiology and Immunology (2005) 49:9 (865-869))　　　　　　　　　　　　　　　
Namsraijav Odontsetseg, Yoshihiro Sakoda, Hiroshi Kida
Microbiology and Immunology, 49, 11, 1017, 1018, 2005年
英語, 研究論文（学術雑誌）

Serological evidence of the persistence of infection with Leptospira interrogans serovar Hardjo in cattle in Mongolia
N Odontsetseg, Y Sakoda, H Kida
MICROBIOLOGY AND IMMUNOLOGY, 49, 9, 865, 869, CENTER ACADEMIC PUBL JAPAN, 2005年, ［査読有り］
英語, 研究論文（学術雑誌）, Serum samples were collected randomly from Mongol breed cattle in three geographically distinct provinces of Mongolia. Antibodies specific to Leptospira interrogans serovar Hardjo were detected by microscopic agglutination test (MAT) at titres of 100 or more in 80.4% (86/107) of the samples from Dornod Province, but in only 28.9% (13/45) in Arkhangai and 23.5% (12/51) in Khuvsgul Provinces, respectively. Treatment of 9 serum samples from Dornod, positive to serovar Hardjo in MAT and negative in the homologous immunoglobulin G (IgG) enzyme-linked immunosorbent assay (ELISA), with 2-mercaptoethanol (2-ME) indicated that those animals were recently infected.

Evaluation of the ESPLINE (R) INFLUENZA A&B-N kit for the diagnosis of avian and swine influenza
GR Bai, Y Sakoda, AS Mweene, N Kishida, T Yamada, H Minakawa, H Kida
MICROBIOLOGY AND IMMUNOLOGY, 49, 12, 1063, 1067, CENTER ACADEMIC PUBL JAPAN, 2005年, ［査読有り］
英語, 研究論文（学術雑誌）, The ESPLINE(R) INFLUENZA A&B-N kit was evaluated for its applicability to the rapid diagnosis of influenza in chickens and pigs. The kit specifically detected viral antigens in tracheal swabs and tissue homogenates of the trachea, liver, spleen, and colon of chickens inoculated with a highly pathogenic avian influenza virus strain, A/chicken/Yamaguchi/7/04 (H5N1), at 48 hr post-inoculation (p.i.) as well as in the tracheal and cloacal swabs and tissue homogenates of dead chickens. For those infected with a low pathogenic strain, A/chicken/aq-Y-55/01 (H9N2), antigens were detected only in the samples from tracheal swabs and organs 1-4 days p.i. The kit also detected viral antigens in the nasal swabs of miniature pigs infected with swine and avian influenza viruses. The kit was found to be sensitive and specific enough for the rapid diagnosis of infections of influenza A virus in chickens and pigs.

Involvement of extracellular signal-regulated kinase and p38 in human diabetic nephropathy
Norihiko Sakai, Takashi Wada, Kengo Furuichi, Yasunori Iwata, Keiichi Yoshimoto, Kiyoki Kitagawa, Satoshi Kokubo, Motoo Kobayashi, Akinori Hara, Junya Yamahana, Toshiya Okumura, Kazuya Takasawa, Shin-Ichi Takeda, Mitsuhiro Yoshimura, Hiroshi Kida, Hitoshi Yokoyama
American Journal of Kidney Diseases, 45, 1, 54, 65, 2005年01月, ［査読有り］
英語, 研究論文（学術雑誌）, Background: The involvement of mitogen-activated protein kinase (MAPK) in human diabetic nephropathy has not been fully investigated. Methods: The presence of cells positive for the phosphorylated MAPK family (phosphorylated extracellular signal-regulated kinase [p-ERK], phosphorylated p38MAPK [p-p38MAPK]) was investigated immunohistochemically in kidneys of 30 patients with diabetic nephropathy. In addition, 10 patients with minimal change nephrotic syndrome, 10 patients with thin basement membrane disease, and 5 patients with benign nephrosclerosis were studied as disease controls. The presence of activated nuclear factor-κB (p65)-positive cells also was evaluated in kidney specimens. Results: In patients with diabetic nephropathy, p-ERK, p-p38MAPK, and p65 were observed in mesangial cells, endothelial cells, podocytes, tubular epithelial cells, and mononuclear infiltrates in interstitium. Numbers of p-ERK-, p-p38MAPK-, and p65-positive cells in both glomeruli and interstitium in patients with diabetic nephropathy were higher than those in controls. In particular, the number of glomerular p-ERK-positive cells in patients with diabetic nephropathy increased in accordance with the progression of glomerular lesions and correlated well with the number of glomerular p65-positive cells (r = 0.654
P <
0.01
n = 30). Conversely, the number of p-p38MAPK-positive cells in glomeruli did not correlate with glomerular lesions. However, the number of tubulointerstitial p-p38MAPK-positive cells in patients with diabetic nephropathy reflected the severity of tubulointerstitial lesions, and numbers of those in the interstitium increased with good correlation to numbers of tubulointerstitial p65-positive cells (r = 0.757
P <
0.01
n = 30) and interstitial CD68-positive macrophages (r = 0.647
P <
0.05
n = 30) and urinary monocyte chemoattractant protein-1 levels (r = 0.605
P <
0.05
n = 30). Conclusion: These results suggest that MAPK phosphorylation contributes to human diabetic nephropathy. In particular, ERK and p38MAPK may be distinctly involved in glomerular and tubulointerstitial lesions in human diabetic nephropathy. © 2004 by the National Kidney Foundation, Inc.

Genetic conservation of hemagglutinin gene of H9 influenza virus in chicken population in Mainland China
JH Liu, K Okazaki, A Mweene, WM Shi, QM Wu, JL Su, GZ Zhang, GR Bai, H Kida
VIRUS GENES, 29, 3, 329, 334, KLUWER ACADEMIC PUBL, 2004年12月, ［査読有り］
英語, 研究論文（学術雑誌）, The hemagglutinin (HA) genes of 12 H9N2 influenza virus strains isolated from chickens in Mainland China during the period 1995 - 2002 were genetically analyzed. All the isolates possessed the same amino acid motif - R-S-S-R/G-L- at the cleavage site of HA. Except for the conserved amino acids, as is the case in the other avian influenza viruses, located in the receptor binding site, all of the 12 isolates possessed N at amino acid position 183; A, T, or V at position 190; K at position 137, whereas the representative strains of the other lineage ( except Dk/HK/Y280/97-like lineage) virus of H9N2 viruses had H, E, and R at these positions respectively. These could be considered as the partial molecular markers of the H9 viruses isolated from chickens in Mainland China. Phylogenetic analyses showed HA genes of these isolates belonged to that of A/duck/Hong Kong/Y280/97-like virus lineage. No A/quail/Hong Kong/Gl/97-like virus was found in chicken, population since the outbreak of H9N2 influenza in Mainland China in 1992. The available evidence indicates that HA genes of H9 influenza virus circulating in Mainland China during the past years were well conserved.

Co-infection of Staphylococcus aureus or Haemophilus paragallinarum exacerbates H9N2 influenza A virus infection in chickens
N Kishida, Y Sakoda, M Eto, Y Sunaga, H Kida
ARCHIVES OF VIROLOGY, 149, 11, 2095, 2104, SPRINGER WIEN, 2004年11月, ［査読有り］
英語, 研究論文（学術雑誌）, H9N2 influenza viruses are frequently isolated from chicken meat and bone marrow imported from China to Japan since 2001. These isolates were experimentally inoculated into specific pathogen-free chickens intranasally. Viruses were recovered from the meat and bone marrow of birds showing no overt signs. On the other hand, chickens co-infected with H9N2 virus and either Staphylococcus aureus or Haemophilus paragallinarum showed clinical signs severer than those shown by birds infected only with the virus alone or each of the bacteria alone. In addition, H9N2 viruses were more efficiently recovered from the chickens co-infected with S. aureus or H. paragallinarum than those from the birds infected with only the virus. The present results indicate that co-infection of H9N2 influenza virus with S. aureus or H. paragallinarum enhances the replication of the virus in chickens, resulting in exacerbation of the H9N2 virus infection.

【高病原性鳥インフルエンザ】インフルエンザウイルスの病原性の分子基盤　　　　　　　　　　　　　　　
村本 裕紀子, 喜田 宏, 河岡 義裕
化学療法の領域, 20, 11, 1663, 1667, (株)医薬ジャーナル社, 2004年10月
日本語, 2003年末から2004年にかけて,高病原性鳥インフルエンザがアジア諸国で流行した.ウイルスはニワトリだけでなくヒトにも感染し,20人以上の死者が出た.致死性の高病原性鳥インフルエンザウイルスは,毎年ヒトの間で流行するインフルエンザウイルスと何が違うのだろうか?本稿では,高病原性鳥インフルエンザウイルスを中心に,インフルエンザウイルスの病原性発現の分子基盤について概説する(著者抄録)

Interregional transmission of the internal protein genes of H2 infoluenza virus in migratory ducks from north America to Eurasia
JH Liu, K Okazaki, GR Bai, WM Shi, A Mweene, H Kida
VIRUS GENES, 29, 1, 81, 86, KLUWER ACADEMIC PUBL, 2004年08月, ［査読有り］
英語, 研究論文（学術雑誌）, H2 influenza virus caused a pandemic in 1957 and has the possibility to cause outbreaks in the future. To assess the evolutionary characteristics of H2 influenza viruses isolated from migratory ducks that congregate in Hokkaido, Japan, on their flyway of migration from Siberia in 2001, we investigated the phylogenetic relationships among these viruses and avian and human viruses described previously. Phylogenetic analysis showed that the PB2 gene of Dk/Hokkaido/107/01 (H2N3) and the PA gene of Dk/Hokkaido/95/01 (H2N2) belonged to the American lineage of avian virus and that the other genes of the isolates belonged to the Eurasian lineage. These results indicate that the internal protein genes might be transmitted from American to Eurasian avian host. Thus, it is further confirmed that interregional transmission of influenza viruses occurred between the North American and Eurasian birds. The fact that reassortants could be generated in the migratory ducks between North American and Eurasian avian virus lineage further stresses the importance of global surveillance among the migratory ducks.

A synthetic peptide from a heptad repeat region of herpesvirus glycoprotein B inhibits virus replication
K Okazaki, H Kida
JOURNAL OF GENERAL VIROLOGY, 85, 8, 2131, 2137, SOC GENERAL MICROBIOLOGY, 2004年08月, ［査読有り］
英語, 研究論文（学術雑誌）, Glycoprotein B (gB) is the most conserved glycoprotein of herpesviruses and plays important roles in virus infectivity. Two intervening heptad repeat (HR) sequences were found in the C-terminal half of all herpesvirus gBs analysed. A synthetic peptide derived from the HR region (aa 477-510) of bovine herpesvirus type 1 (BoHV-1) gB was studied for its ability to inhibit virus replication. The peptide interfered with cell-to-cell spread and consistently inhibited replication of BoHV-1, with a 50% effective concentration value (EC50) of 5 muM. Inhibition of replication was obtained not only with herpesviruses including pseudorabies virus and herpes simplex virus type 1 but also partly with Newcastle disease virus. Possible mechanisms of membrane fusion inhibition by the peptide are discussed.

[Avian influenza virus].
Kida H
Uirusu, 54, 1, 93, 96, 1, 2004年06月, ［査読有り］
日本語, 2003年から2004年にかけて日本を含むアジア9カ国で発生した一連の高病原性鳥インフルエンザは, 感染症に国境がないこと, 日本の鶏群にも本症発生のリスクがあることに加えてウイルスの侵入をいち早く検出するために年間を通した家禽のモニタリングが必須であることを改めて諭す教訓となった. アジアでは今なお高病原性鳥H5N1インフルエンザウイルスによる家禽の被害が断続している. さらにタイとベトナムではそれぞれ12名および22名のヒトがH5N1ウイルスに感染し, 8名および15名が死亡した. 今後, 再び高病原性鳥インフルエンザが日本に飛び火する可能性を否定できない状況にある. 家禽における鳥インフルエンザウイルスの感染を早期に摘発, 淘汰することによって家禽の被害を最小限にくい止めるとともに, ヒトの健康と食の安全を守ることが鳥インフルエンザ対策の基本である.

[Round table discussion on highly pathogenic H5N1 avian influenza].
Nagai Y, Otsuki K, Abe N, Kawaoka Y, Kida H, Kurata T, Sata T, Tashiro M, Yamaguchi N, Yoshikura H
Uirusu, 54, 123, 141, 1, 2004年06月, ［査読有り］

Ｈ５強毒インフルエンザウイルスの神経病原性と解熱剤投与の影響に関する実験的研究　　　　　　　　　　　　　　　
梅村孝司, 松田一哉, 朴天鍋, 寸田祐嗣, 木村享史, 落合謙爾, 喜田宏
日本病理学会会誌, 93, 1, 175, 2004年04月
日本語, 研究論文（学術雑誌）

A型インフルエンザウイルスPB2遺伝子分節のパッケージングと粒子形成に必要な領域　　　　　　　　　　　　　　　
村本 裕紀子, 高田 礼人, 藤井 健, 堀本 研子, 渡辺 真治, 喜田 宏, 河岡 義裕
日本獣医学会学術集会講演要旨集, 137回, 91, 91, (公社)日本獣医学会, 2004年03月
日本語

Experimental dual infection of pigs with an H1N1 swine influenza virus (A/Sw/Hok/2/81) and Mycoplasma hyopneumoniae
S Yazawa, M Okada, M Ono, S Fujii, Y Okuda, Shibata, I, H Kida
VETERINARY MICROBIOLOGY, 98, 3-4, 221, 228, ELSEVIER SCIENCE BV, 2004年03月, ［査読有り］
英語, 研究論文（学術雑誌）, Dual infection of pigs with swine influenza virus (SIV) and Mycoplasma hyopneumoniae was carried out to compare the clinical and pathological effects of dual infection in caesarian derived and colostrums deprived (CDCD) pigs, with that of a single infection with M. hyopneumoniae. In Experiment 1, 40-day-old CDCD pigs were inoculated only with SIV (A/Sw/Hok/2/81, H1N1). The virus was isolated from nasal swabs for 5-6 days. None of these pigs showed clinical signs of infection throughout the experimental period. These results suggested that this strain can infect pigs but is only slightly pathogenic when it is inoculated singly to a CDCD pig. In Experiment 2, 60-day-old CDCD pigs were inoculated with M. hyopneumoniae and then were inoculated with SIV (A/Sw/Hok/2/8 1) at 1 week (MHYO-7d-SIV-7d group) or 3 weeks (MHYO-21 d-SIV-7d group) after M. hyopneumoniae inoculation. Macroscopically, dark red-to-purple lung lesions were observed in all of pigs at 14 or 28 days post-inoculation. Percentages of dark red-to-purple lung lesions in dual infection groups (MHYO-7d-SIV-7d group: 18.7 +/- 4.2%, MHYO-21 d-SIV-7d group: 23.0 +/- 8.0%) were significantly (P < 0.05) increased compared to those of each control group in which pigs were inoculated only with M. hyopneumoniae (MHYO-14d group: 4.7 +/- 2.9%, MHYO-28 group: 3.3 +/- 2.4%). Microscopically, bronchial epithelial lesions (epithelial disruption, degeneration, hyperplasia and formation of microabscess) were frequently observed in dark red-to-purple lung lesions of only the dual infection groups. These results demonstrate that the lung lesion of pigs inoculated with M. hyopneumoniae and SIV is more severe than that of pigs inoculated only with M. hyopneumoniae. (C) 2003 Elsevier B.V. All rights reserved.

The vagus nerve is one route of transneural invasion for intranasally inoculated influenza a virus in mice.
Matsuda K, Park CH, Sunden Y, Kimura T, Ochiai K, Kida H, Umemura T
Veterinary pathology, 41, 101, 107, 2, 2004年03月, ［査読有り］

Antibodies to human-related H3 influenza a virus in Baikal seals (Phoca sibirica) and ringed seals (Phoca hispida) in Russia
K Ohishi, N Kishida, A Ninomiya, H Kida, Y Takada, N Miyazaki, AN Boltunov, T Maruyama
MICROBIOLOGY AND IMMUNOLOGY, 48, 11, 905, 909, WILEY-BLACKWELL, 2004年, ［査読有り］
英語, 研究論文（学術雑誌）, Antibodies to influenza A virus were detected using enzyme-linked immunosorbent assay (ELISA) in the sera from two of seven Baikal seals (Phoca sibrica) and from five of six ringed seals (Phoca hispida) in Russia. In a hemagglutination-inhibition test using H1-H15 reference influenza A viruses, ELISA-positive sera from one Baikal seal and four ringed seals reacted to A/Aichi/2/68 (H3N2) and A/Bangkok/1/79 (H3N2) strains. One ringed seal serum sample reacted to A/seal/Massachusetts/1/80 (H7N7). The present results suggested that human-related H3 viruses were prevalent in Baikal seals and ringed seals inhabiting the central Russian Arctic.

Pathologic findings of initial biopsies reflect the outcomes of membranous nephropathy
Keiichi Yoshimoto, Hitoshi Yokoyama, Takashi Wada, Kengo Furuichi, Norihiko Sakai, Yasunori Iwata, Satoshi Goshima, Hiroshi Kida
Kidney International, 65, 1, 148, 153, Blackwell Publishing Inc., 2004年, ［査読有り］
英語, 研究論文（学術雑誌）, Background. A considerable diversity of prognosis is seen with idiopathic membranous nephropathy (IMN). The initial factors affecting long-term outcome remain unclear. Methods. We studied retrospectively 105 patients with IMN who had been followed up for at least 5 years, or until end-stage renal failure (ESRF) (primary outcome), or death (secondary outcome). We analyzed the initial clinicopathologic factors affecting primary and secondary outcomes. We assigned the patients to two groups and one subgroup, based on the electron microscopic findings. The groupings were: homogeneous type with synchronous electron dense deposits
homogeneous type with large dense deposits (deep subgroup)
and heterogeneous type with various phases of dense deposits. Results. No differences in the initial clinicopathologic states were seen between the homogeneous (N = 60) and heterogeneous types (N = 45), apart from hypertension and disease history before biopsy. In the homogeneous type, only one patient developed ESRF, which was drug-induced, and remission occurred earlier than in the heterogeneous type. With regard to secondary outcomes, increased age, male gender, heterogeneous type, and deep subgroup were independent risk factors. There were no significant differences attributable to therapeutic regime with respect to primary or secondary outcome in either group. Conclusion. Our results indicate that an electron microscopic classification, at initial biopsy, as heterogeneous type or deep subgroup type with dense deposits are independent indicators of poor prognosis in IMN.

Induction of interferon-inducible protein-10 and monokine induced by interferon-gamma from human endothelial cells infected with Influenza A virus
N Ishiguro, A Takada, M Yoshioka, Ma, X, H Kikuta, H Kida, K Kobayashi
ARCHIVES OF VIROLOGY, 149, 1, 17, 34, SPRINGER-VERLAG WIEN, 2004年01月, ［査読有り］
英語, 研究論文（学術雑誌）, Primary human umbilical vein endothelial cells (HUVECs) were infected with Influenza virus A/Aichi/2/68 (H3N2) in order to determine the role of endothelial cells in mediating inflammation induced upon virus infection. Structural proteins of the virus and mRNA of the M2 protein were detected in the infected cells, indicating that virus infection had occurred in HUVECs. The Influenza A virus-infected HUVECs showed elevated levels of gene expression of interferon (IFN)-inducible protein (IP)-10 and monokine induced by IFN-gamma (Mig), while heat-, formalin- and diethyl ether-inactivated viruses did not enhance the IP-10 and Mig gene expression. The results thus indicate that infection of live Influenza A virus is responsible for elevation of IP-10 and Mig gene expression. The elevation of IP-10 and Mig gene expression in infected HUVECs was not accompanied by the elevation of IFN-gamma gene expression, indicating that the elevation of IP-10 and Mig gene expression was independent of the IFN-gamma pathway.

Intranasal immunization with formalin-inactivated virus vaccine induces a broad spectrum of heterosubtypic immunity against influenza virus infection in mice
A Takada, S Matsushita, A Ninomiya, Y Kawaoka, H Kida
OPTIONS FOR THE CONTROL OF INFLUENZA V, 1263, 618, 621, ELSEVIER SCIENCE BV, 2004年, ［査読有り］
英語, 研究論文（国際会議プロシーディングス）, It has been known that the influenza A virus infection induces a cross-protective immunity against infection by viruses with different subtypes of viral envelope proteins, hemagglutinin and neuraminidase. This heterosubtypic immunity is generally mediated by cytotoxic T lymphocytes (CTL) reactive to specific epitopes in the viral internal proteins, such as nucleoprotein and matrix protein. By contrast, immunization with inactivated virus antigens has been thought to be unable to generate heterosubtypic immunity, since inactivated antigens do not usually induce CTL responses. However, we show that intranasal immunization with formalin-inactivated intact virus, but not ethersplit vaccines, induced a broad spectrum of heterosubtypic protective immunity in mice. The protection may be mediated by the mucosal immune response, most likely secretory IgA antibodies to the viral proteins. This approach may overcome limitations in the efficacy of inactivated influenza vaccines and confer potent immunity to humans against viruses with new pandemic potential. (C) 2004 Elsevier B.V. All rights reserved.

Enhancement of pathogenicity of H9N2 influenza A viruses isolated from chicken in China by co-infection with Staphylococcus aureus and Haemophilus paragallinarum
N Kishida, M Eto, Y Sunaga, H Kida
OPTIONS FOR THE CONTROL OF INFLUENZA V, 1263, 481, 485, ELSEVIER SCIENCE BV, 2004年, ［査読有り］
英語, 研究論文（国際会議プロシーディングス）, H9N2 influenza viruses were isolated from the meat and bone marrow of chickens imported from China to Japan in 2001. These isolates were experimentally inoculated intranasally into specific, pathogen-free chickens. Viruses were recovered from the meat and bone marrow of the birds without any clinical signs. On the other hand, chickens co-infected with the H9N2 virus and with either Staphylococcus aureus or Haemophilus paragallinarum showed clinical signs much severer than those shown by chickens infected only with S. aureus or H. paragallinarum. Higher titers of H9N2 viruses were recovered from the chickens co-infected with S. aureus or H. paragallinarum compared with those from birds infected only with the virus. The present results indicate that co-infection with S. aureus or H. paragallinarum enhances the replication of the H9N2 virus, resulting in exacerbation of the disease. (C) 2003 Elsevier B.V. All rights reserved.

Preparation of a panel of avian influenza viruses of different subtypes for vaccine strains against future pandemics.　　　　　　　　　　　　　　　
Sakoda Y, Okazaki K, Takada A, Ito Y, Tamai K, Okamatsu M, Ito T, Shortridge KF, Webster RG, Kida H
Options for the Control of Influenza V, International congress series, 1263, 674, 677, 2004年, ［査読有り］
英語, 研究論文（学術雑誌）

Phylogenetic analysis of hemagglutinin and neuraminidase genes of H9N2 viruses isolated from migratory ducks
JH Liu, K Okazaki, WM Shi, H Kida
VIRUS GENES, 27, 3, 291, 296, KLUWER ACADEMIC PUBL, 2003年12月, ［査読有り］
英語, 研究論文（学術雑誌）, Genetic analysis indicated that the pandemic influenza strains derived from wild aquatic birds harbor viruses of 15 hemagglutinin (HA) and 9 neuraminidase (NA) antigenic subtypes. Surveillance studies have shown that H9N2 subtype viruses are worldwide in domestic poultry and could infect mammalian species, including humans. Here, we genetically analyzed the HA and NA genes of five H9N2 viruses isolated from the migratory ducks in Hokkaido, Japan, the flyway of migration from Siberia during 1997-2000. The results showed that HA and NA genes of these viruses belong to the same lineages, respectively. Compared with those of A/quail/Hong Kong/G1/97-like and A/duck/Hong KongIY280/97-like viruses, HA and NA of the migratory duck isolates had a close relationship with those of H9N2 viruses isolated from the chicken in Korea, indicating that the Korea H9N2 viruses might be derived from the migratory ducks. The NA genes of the five isolates were located in the same cluster as those of N2 viruses, which had caused a human pandemic in 1968, indicating that the NA genes of the previous pandemic strains are still circulating in waterfowl reservoirs. The present results further emphasize the importance of carrying out molecular epidemiological surveillance of H9N2 viruses in wild ducks to obtain more information for the future human influenza pandemics preparedness.

Persistence of viral RNA segments in the central nervous system of mice after recovery from acute influenza A virus infection
Park CH, Matsuda K, Sunden Y, Ninomiya A, Takada A, Ito H, Kimura T, Ochiai K, Kida H, Umemura T
Veterinary Microbiology, 97, 259, 268, 2003年12月, ［査読有り］
英語, 研究論文（学術雑誌）

[Distribution and circulation of influenza viruses in nature].
Kida H
Nihon rinsho. Japanese journal of clinical medicine, 61, 1865, 1871, 11, 2003年11月, ［査読有り］

The effects of antipyretics on influenza virus encephalitis in mice and chicks
Y Sunden, CH Park, K Matsuda, A Anagawa, T Kimura, K Ochiai, H Kida, T Umemura
JOURNAL OF VETERINARY MEDICAL SCIENCE, 65, 11, 1185, 1188, JAPAN SOC VET SCI, 2003年11月, ［査読有り］
英語, 研究論文（学術雑誌）, To investigate the effect of antipyretics on the murine and poultry models of influenzal encephalitis, we injected large quantities of antipyretics, acetylsalicylic acid (aspirin) and dicrofenac sodium (voltaren). The effect of antipyretic treatment on the murine encephalitis model was unremarkable histologically and immunohistochemically. Whereas in chicks, CNS lesions consisting of perivascular cuffing and gliosis appeared only in those animals treated with the antipyretics and viral antigen was detected mainly in the nuclei of glial cells or vascular endothelia of voltaren-treated animals. We here demonstrate that antipyretic treatment aggravated the hematogenous spread of the influenza virus to the CNS in chicks, but did not affect the transneural infection in mice.

Phylogenetic analysis of neuraminidase gene of H9N2 influenza viruses prevalent in chickens in China during 1995-2002
JH Liu, K Okazaki, WM Shi, QM Wu, AS Mweene, H Kida
VIRUS GENES, 27, 2, 197, 202, KLUWER ACADEMIC PUBL, 2003年10月, ［査読有り］
英語, 研究論文（学術雑誌）, The neuraminidase (NA) genes of 12 H9N2 influenza virus strains isolated from diseased chickens in different farms in mainland China during 1995-2002 were amplified and sequenced. Amino acids at hemadsorbing (HB) site of these isolates are different from those of A/quail/Hong Kong/G1/97-like viruses and A/chicken/Korea/96-like viruses. Neuraminidases of the 12 strains had a deletion of 3 amino acid residues at positions 63-65 as compared to that of A/turkey/Wisconsin/189/66, while those of Korea and Pakistan H9N2 isolates had no deletion. Phylogenetic analyses showed NA gene of these isolates belonged to that of A/duck/Hong Kong/Y280/97-like virus lineage. NA gene of the H9N2 viruses isolated in Korea and Pakistan belonged to lineage different from those of the 12 isolates. The present results indicate that the NA of H9N2 strains isolated in mainland China during the past 8 years were well preserved and the geographical distribution play a significant role in the evolution of the H9N2 influenza viruses.

H9N2 influenza viruses prevalent in poultry in China are phylogenetically distinct from A/quail/Hong Kong/G1/97 presumed to be the donor of the internal protein genes of the H5N1 Hong Kong/97 virus
JH Liu, K Okazaki, H Ozaki, Y Sakoda, QM Wu, FY Chen, H Kida
AVIAN PATHOLOGY, 32, 5, 551, 560, CARFAX PUBLISHING, 2003年10月, ［査読有り］
英語, 研究論文（学術雑誌）, Ten H9N2 influenza virus strains isolated from diseased chickens in different farms in China during 1995 to 1999 were antigenically and genetically characterized. The haemagglutinins of the isolates were not related to those of A/quail/ Hong Kong/G1/97 ( H9N2) (Qa/HK/G1/97), but were closely related to that of A/chicken/ Hong Kong/G9/97 ( H9N2) (Ck/HK/G9/97). The neuraminidase of these isolates had a deletion of three amino acid residues at positions 63 to 65 as compared with those of Ck/HK/G9/97, while that of Qa/HK/G1/97 lacked two amino acids at positions 38 and 39. The PB2 genes of the isolates were not related to those of Qa/HK/G1/ 97 or Ck/HK/G9/97, but showed some relationship to that of A/duck/Hong Kong/Y439/97 ( H9N2) (Dk/HK/Y439/ 97). The PB1 genes of the isolates were not related to those of the three representative strains. The PA, NP, M, and NS genes of the isolates belonged to the same lineage as those of Ck/HK/G9/97, and were distinct from those of Qa/HK/G1/ 97 and Dk/HK/Y439/97. The present results indicate that H9N2 influenza viruses prevalent in chicken populations in China belong genetically to one lineage and are distinct from Qa/HK/G1/ 97, presumed to be the donor of the internal protein genes of the highly pathogenic H5N1 influenza virus in Hong Kong in 1997.

Neurotropism of the 1997 Hong Kong H5N1 influenza virus in mice
H Tanaka, CH Park, A Ninomiya, H Ozaki, A Takada, T Umemura, H Kida
VETERINARY MICROBIOLOGY, 95, 1-2, 1, 13, ELSEVIER SCIENCE BV, 2003年08月, ［査読有り］
英語, 研究論文（学術雑誌）, The direct transmission of H5N1 influenza A viruses from chickens to humans in Hong Kong in 1997 emphasized the need to have information on the pathogenesis of avian influenza virus infection in mammals. H5N1 influenza viruses isolated from patients during the incident killed experimentally infected mice. The principal lesions of the mice were broncho-interstitial pneumonia and nonsuppurative encephalitis. Infectious viruses and/or viral antigens were detected in the brain as well as in the trigeminal and vagal ganglia but not in the blood of the mice. These findings suggest that the virus reached the brain through the vagus and/or trigeminal nerves following replication in the respiratory mucosa. The results imply that neurotropism of the H5N1 virus in mice is a novel characteristic in the pathogenesis of infection by human influenza virus isolates. (C) 2003 Elsevier B.V. All rights reserved.

Intranasal immunization with formalin-inactivated virus vaccine induces a broad spectrum of heterosubtypic immunity against influenza A virus infection in mice
A Takada, S Matsushita, A Ninomiya, Y Kawaoka, H Kida
VACCINE, 21, 23, 3212, 3218, ELSEVIER SCI LTD, 2003年07月, ［査読有り］
英語, 研究論文（学術雑誌）, It has been known that influenza A virus infection induces a cross-protective immunity against infection by viruses with different subtypes of viral envelope proteins, hemagglutinin (HA) and neuraminidase (NA). This heterosubtypic immunity is generally mediated by cytotoxic T lymphocytes (CTL) reactive to specific epitopes in the viral internal proteins, such as nucleoprotein and matrix protein. By contrast, immunization with inactivated virus antigens has been thought to be unable to generate heterosubtypic immunity, since inactivated antigens do not usually induce CTL responses. However, we show that intranasal immunization with formalin-inactivated intact virus, but not ether-split vaccines, induced a broad spectrum of heterosubtypic protective immunity in mice. The protection may be mediated by the mucosal immune response, most likely secretory IgA antibodies to the viral proteins. This approach may overcome limitations in the efficacy of inactivated influenza vaccines and confer potent immunity to humans against viruses with new pandemic potential. (C) 2003 Elsevier Science Ltd. All rights reserved.

[For the prevention and control of pandemic influenza].
Kida H
Uirusu, 53, 1, 71, 74, 1, 2003年06月, ［査読有り］
日本語, 1968年に出現した新型香港Aインフルエンザウイルス A/Hong Kong/68 (H3N2) 株は, カモがシベリアの営巣湖沼から家禽 (アヒル) に持ち込んだウイルスと当時ヒトに流行していたH2N2ウイルスが, 中国南部でブタの呼吸器に共感染して生じた遺伝子再集合体である. 1957年の新型H2N2アジアインフルエンザウイルスも同様の経路で出現したものと推定される.1918年に出現して2千万以上の人命を奪ったH1N1スペインインフルエンザウイルスが出現した経路について, これまでに報告された成績から次の考察が導かれる. すなわち, このウイルス株のHAおよびNA遺伝子は, カモがカナダの営巣湖沼から持ち込んだウイルスに起源がある. カモが糞便とともに排泄したウイルスは, 米国の家禽 (シチメンチョウかアヒル) に水系感染し, さらにブタに伝播した. ブタは当時ヒトに流行していたH3N8インフルエンザウイルスにも共感染して, HAおよびNA遺伝子がカモのウイルスに由来する遺伝子再集合体を産生した. これがまずイリノイ州でブタにインフルエンザの流行を起こし, 忽ち米国中西部一帯に広がった. その間にヒトに感染伝播したものがスペインインフルエンザウイルス株である.1997年, 香港で18人に感染して6人を死亡させたニワトリのH5N1ウイルスもまた, シベリアからカモが家禽に持ち込み, ニワトリに受け継がれる間に病原性を獲得したものである. すなわち, 新型ウイルスの遺伝子はすべてカモの腸内ウイルスに由来する. 実際, 家畜, 家禽, 野生鳥獣およびヒトのインフルエンザウイルス遺伝子の系統解析成績は, 全てがカモのウイルスに起源があることを示している.カモはすべてのHA (H1-H15) とNA (N1-N9) 亜型のインフルエンザウイルスを保有する. カモは北方の営巣湖沼でインフルエンザウイルスに水系経口感染し, 結腸陰窩の上皮細胞で増殖したウイルスを糞便と共に湖沼水中に排泄する. 秋になると, カモは南方に渡る. カモが不在の冬期, ウイルスは湖沼水中に凍結保存される. ヒトのウイルスとカモのウイルスがブタに同時感染すると, 両ウイルスの遺伝子再集合体が生ずる. 過去に出現した新型ウイルス遺伝子の導入経路は, 上に述べたように, カモ→アヒル (家禽) →ブタ→ヒトである.1996年秋にシベリアから北海道に飛来したカモから分離した弱毒H5N4ウイルスを不活化して試製したワクチンは, 1997年に香港でニワトリとヒトから分離された強毒H5N1ウイルスに対する力価試験に合格した. また, このワクチンを鼻腔内に滴下したマウスは, 強毒H5N1ウイルスの攻撃に耐過した. すなわち, カモから分離される弱毒ウイルスの中から, 適切な株を予め選定しておけば, 新型ウイルスの出現に際して, 迅速にワクチンを準備できることが判った.以上のように, 新型インフルエンザウイルスはカモの腸内ウイルスが家禽を経て, あるいはさらにブタの呼吸器でヒトのウイルスの遺伝子を獲得した再集合体として出現する. 新型ウイルスの出現に備え, 水禽, その営巣湖沼水, 家禽, ブタとヒトの疫学調査を強化し, 新型ウイルスの出現とその亜型を予測すると共に, 調査で分離されるウイルスの中からワクチン候補株ならびに診断抗原を選出し, 系統保存・供給するグローバルサーベイランス計画"Programme of Excellence for the Control of Pandemic Influenza"を推進している.

[Report of the 50th annual meeting of the Japanese Society for Virology].
Kida H
Uirusu, 53, 1, 93, 94, 1, 2003年06月, ［査読有り］
日本語

Replication and transmission of influenza viruses in Japanese quail
NV Makarova, H Ozaki, H Kida, RG Webster, DR Perez
VIROLOGY, 310, 1, 8, 15, ACADEMIC PRESS INC ELSEVIER SCIENCE, 2003年05月, ［査読有り］
英語, 研究論文（学術雑誌）, Quail have emerged as a potential intermediate host in the spread of avian influenza A viruses in poultry in Hong Kong. To better understand this possible role, we tested the replication and transmission in quail of influenza A viruses of all 15 HA subtypes. Quail supported the replication of at least 14 subtypes. Influenza A viruses replicated predominantly in the respiratory tract. Transmission experiments suggested that perpetuation of avian influenza viruses in quail requires adaptation. Swine influenza viruses were isolated from the respiratory tract of quail at low levels. There was no evidence of human influenza A or B virus replication. Interestingly, a human-avian recombinant containing the surface glycoprotein genes of a quail virus and the internal genes of a human virus replicated and transmitted readily in quail; therefore, quail could function as amplifiers of influenza virus reassortants that have the potential to infect humans and/or other mammalian species. (C) 2003 Elsevier Science (USA). All rights reserved.

強毒インフルエンザウイルスは血液凝固不全を引き起こす　　　　　　　　　　　　　　　
村本 裕紀子, 尾崎 弘一, 高田 礼人, 朴 天鎬, 寸田 祐嗣, 梅村 孝司, 河岡 義裕, 喜田 宏
日本獣医学会学術集会講演要旨集, 135回, 128, 128, (公社)日本獣医学会, 2003年03月
日本語

中国産輸入家きん肉からのニューカッスル病ウイルス及びH9N2亜型インフルエンザウイルスの分離と分離株の性状　　　　　　　　　　　　　　　
衛藤 真理子, 真瀬 昌司, 岸田 典子, 米川 和宏, 高橋 周子, 喜田 宏, 須永 裕
日本獣医学会学術集会講演要旨集, 135回, 127, 127, (公社)日本獣医学会, 2003年03月
日本語

Insertion of cellular sequence and RNA recombination in the structural protein coding region of cytopathogenic bovine viral diarrhoea virus
M Nagai, Y Sakoda, M Mori, M Hayashi, H Kida, H Akashi
JOURNAL OF GENERAL VIROLOGY, 84, 2, 447, 452, SOC GENERAL MICROBIOLOGY, 2003年02月, ［査読有り］
英語, 研究論文（学術雑誌）, The cytopathogenic bovine viral diarrhoea virus (cp BVDV) strain KS86-1 cp was isolated from a calf persistently infected with the noncytopathogenic (ncp) strain KS86-1 ncp after it was exposed to cp BVDV strain Nose and developed mucosal disease (MD). Molecular analysis revealed that an insertion of a cellular gene and a duplication of the viral RNA encoding the nucleocapsid protein C and part of N-pro had occurred in the C coding region of the Nose and KS86-1 cp genomes. The inserted cellular gene was closely related to the cINS sequence. Remarkably, the 5' upstream region from the insertion of KS86-1 cp had high sequence identity to that of Nose, but differed from that of KS86-1 ncp. In contrast, the region downstream from the insertion of KS86-1 cp showed high identity to KS86-1 ncp, but not to Nose. These data reveal that KS86-1 cp is a chimeric virus generated by homologous RNA recombination in a calf with MD.

JC virus strains indigenous to northeastern Siberians and Canadian Inuits are unique but evolutionally related to those distributed throughout Europe and Mediterranean areas
C Sugimoto, M Hasegawa, HY Zheng, Demenev, V, Y Sekino, K Kojima, T Honjo, H Kida, T Hovi, T Vesikari, JA Schalken, K Tomita, Y Mitsunobu, H Ikegaya, N Kobayashi, T Kitamura, Y Yogo
JOURNAL OF MOLECULAR EVOLUTION, 55, 3, 322, 335, SPRINGER-VERLAG, 2002年09月, ［査読有り］
英語, 研究論文（学術雑誌）, Human polyomavirus JC virus (JCV) isolates around the world are classified into more than 10 geographically distinct genotypes (designated as subtypes). Evolutionary relationships among JCV subtypes were recently examined, and the following pattern of JCV evolution was indicated. The ancestral JCV first divided into three superclusters, designated Types A, B, and C. A split in Type A generated two subtypes, EU-a and -b, containing mainly European and Mediterranean isolates. The split in Type B generated Af 2 (the major African subtype), B1-c (a minor European subtype), and various Asian subtypes. Type C generated a single subtype (Afl), consisting of isolates derived from western Africa. In this study, JCV isolates prevalent among northeastern Siberians and Canadian Inuits were evaluated in the context of the above-described pattern of JCV evolution. The Siberian/Arctic JCV isolates were classified as belonging mainly to Type A, based on the result of a preliminary phylogenetic analysis. We then examined, using the whole-genome approach, the phylogenetic relationships among worldwide Type A isolates. In neighbor-joining and maximum-likelihood analyses, Type A JCVs worldwide consistently diverged into three subtypes, EU-a, -b, and -c, with high bootstrap probabilities. EU-c was constructed only by northeastern Siberian isolates, derived mainly from Nanais living in the lower Amur River region, and was shown to have been generated by the first split in Type A. Most Siberian/Arctic isolates derived from Chukchis, Koryaks, and Canadian Inuits formed a distinct cluster within the EU-a subtype, with a high bootstrap probability. Based on the present findings, we discuss ancient human migrations, accompanied by Type A JCVs, across Asia and to Arctic areas of North America.

Generation of velogenic Newcastle disease viruses from a nonpathogenic waterfowl isolate by passaging in chickens
Y Shengqing, N Kishida, H Ito, H Kida, K Otsuki, Y Kawaoka, T Ito
VIROLOGY, 301, 2, 206, 211, ACADEMIC PRESS INC ELSEVIER SCIENCE, 2002年09月, ［査読有り］
英語, 研究論文（学術雑誌）, A benign Newcastle disease virus (NDV) recently became highly virulent during replication in domestic chickens. It is still unclear whether NDVs circulating among wild waterfowl also have the potential to become highly pathogenic (velogenic) in chickens. To demonstrate experimentally the generation of velogenic NDV from a nonpathogenic waterfowl isolate, we passaged an avirulent goose isolate in chickens. After nine consecutive passages by air-sac inoculation, followed by five passages in chick brain, the virus became highly virulent in chickens, producing a 100% mortality rate, and demonstrating typical velogenic properties in pathogenicity tests. Sequence analysis at the fusion protein cleavage site showed that the original isolate contained the typical avirulent type sequence, E-R-Q-E-R/L, which progressed incrementally to a typical virulent type, K-R-Q-K-R/F, during repeated passage in chickens. These results demonstrate that avirulent viruses, maintained in wild waterfowl in nature and bearing the consensus avirulent type sequence, have the potential to become velogenic after transmission to and circulation in chicken populations. The results also suggest that chickens provide a mechanism for the selection of virulent viruses from an avirulent background. (C) 2002 Elsevier Science (USA).

Seroepidemiological evidence of avian H4, H5, and H9 influenza A virus transmission to pigs in southeastern China
A Ninomiya, A Takada, K Okazaki, KF Shortridge, H Kida
VETERINARY MICROBIOLOGY, 88, 2, 107, 114, ELSEVIER SCIENCE BV, 2002年08月, ［査読有り］
英語, 研究論文（学術雑誌）, Pig serum samples collected in southeastern China were examined for antibodies to influenza A viruses. Since the hemagglutination inhibition (HI) test does not accurately detect antibodies to the hemagglutinins (HAs) of "avian" influenza viruses, we utilized the neutralization (NT) test to detect subtype-specific antibodies to the HA of avian viruses in pig sera. Neutralizing antibodies to H1, H3, H4, and H5 influenza viruses were detected in the serum samples collected in 1977-1982 and 1998, suggesting that pigs in China have been sporadically infected with avian H4 and H5 viruses in addition to swine and human HI and H3 viruses. Antibodies to H9 virus, on the other hand, were found only in the sera collected in 1998, not in those collected in 1977-1982, correlating with the recent spread in poultry and subsequent isolation of H9N2 viruses from pigs and humans in 1998. The present results indicate that avian influenza viruses have been transmitted to pig populations in southeastern China. (C) 2002 Elsevier Science B.V. All rights reserved.

Intranasal administration of a synthetic peptide vaccine encapsulated in liposome together with an anti-CD40 antibody induces protective immunity against influenza A virus in mice
A Ninomiya, K Ogasawara, K Kajino, A Takada, H Kida
VACCINE, 20, 25-26, 3123, 3129, ELSEVIER SCI LTD, 2002年08月, ［査読有り］
英語, 研究論文（学術雑誌）, Mucosal immunity is critical for protection from viral infections. We attempted to activate mucosal cytotoxic T lymphocytes (CTLs) specific for influenza A virus nucleoprotem (NP) which play an important role in protective immunity. It has been shown that dendritic cells (DCs) activated by signaling via CD40-CD40 ligand (CD40L) interaction are required for the differentiation of naive CD8(+) T cells into antigen-specific CTLs in a non-mucosal environment. We herein inoculated mice intranasally with an anti-CD40 monoclonal antibody (anti-CD40 mAb) and NP366-374 peptide, corresponding to a CTL epitope on NP, encapsulated in liposome (liposomal NP366-374) to induce protective CTL responses against influenza A virus. Intranasal but not subcutaneous immunization with liposomal NP366-374 effectively induced mucosal immunity to reduce virus replication in the lung, suggesting that anti-CD40 mAb also functioned as a mucosal adjuvant. Interestingly, neither MHC class I- nor class II-deficient mice immunized intranasally with these materials were resistant to the infection. Since anti-CD40 mAb was considered to help replace CD4(+) T cells, another help of CD4(+) T cells are presumably required for the induction of CTL activity in the lung. This approach may prove promising for developing vaccines to induce mucosal CTL responses, and seems to highlight differences between mucosal and non-mucosal immunity. (C) 2002 Elsevier Science Ltd. All rights reserved.

Influenza A virus with defective M2 ion channel activity as a live vaccine
T Watanabe, S Watanabe, H Kida, Y Kawaoka
VIROLOGY, 299, 2, 266, 270, ACADEMIC PRESS INC ELSEVIER SCIENCE, 2002年08月, ［査読有り］
英語, 研究論文（学術雑誌）, We propose a rational approach to the design of live virus vaccines against influenza infection by alteration of the influenza A virus M2 protein, which is responsible for ion channel activity. Previously we demonstrated that a mutant A/WSN/33 (H1N1) influenza virus with defective M2 ion channel activity did not show appreciable growth defects in cell culture, although its growth was attenuated in mice (T Watanabe, S. Watanabe, H. Ito, H. Kida, and Y Kawaoka, 2001, J. Virol. 75, 5656-5662). Here, we show that this M2 ion channel defective mutant virus, the M2del29-31, protected mice against challenge with lethal doses of influenza virus, indicating the potential of incorporating this M2 alteration in a live influenza vaccine as one of the attenuating mutations. (C) 2002 Elsevier Science (USA).

The immediate early gene of canine herpesvirus is transcribed through early and late phases
M Miyoshi, K Okazaki, M Takiguchi, H Kida, A Hashimoto
JOURNAL OF VETERINARY MEDICAL SCIENCE, 64, 7, 627, 631, JAPAN SOC VET SCI, 2002年07月, ［査読有り］
英語, 研究論文（学術雑誌）, The immediate early (IE) gene of canine herpesvirus (CHV), homologue of the infected cell protein 4 (ICP4) gene of herpes simplex virus 1, is transcribed as a 4.9kb mRNA during IE phase. The IE gene was further transcribed as a 4.8kb mRNA through early (E) and late (L) phases of productive infection. Transcription of the 4.8kb mRNA initiated from downstream of the TATA box in an intron which was spliced out during IE phase. The reverse transcription-polymerase chain reaction revealed that the IE promoter was turned off during L phase at a permissive temperature. We, thus, propose to redesignate the IE gene of CHV as CICP4 gene.

The invasion routes of neurovirulent A/Hong Kong/483/97 (H5N1) influenza virus into the central nervous system after respiratory infection in mice.
Park CH, Ishinaka M, Takada A, Kida H, Kimura T, Ochiai K, Umemura T
Archives of virology, 147, 1425, 1436, 7, 2002年07月, ［査読有り］

Ebola virus VP40 drives the formation of virus-like filamentous particles along with GP
T Noda, H Sagara, E Suzuki, A Takada, H Kida, Y Kawaoka
JOURNAL OF VIROLOGY, 76, 10, 4855, 4865, AMER SOC MICROBIOLOGY, 2002年05月, ［査読有り］
英語, 研究論文（学術雑誌）, Using biochemical assays, it has been demonstrated that expression of Ebola virus VP40 alone in mammalian cells induced production of particles with a density similar to that of virions. To determine the morphological properties of these particles, cells expressing VP40 and the particles released from the cells were examined by electron microscopy. VP40 induced budding from the plasma membrane of filamentous particles, which differed in length but had uniform diameters of approximately 65 nm. When the Ebola virus glycoprotein (GP) responsible for receptor binding and membrane fusion was expressed in cells, we found pleomorphic particles budding from the plasma membrane. By contrast, when GP was coexpressed with VP40, GP was found on the filamentous particles induced by VP40. These results demonstrated the central role of VP40 in formation of the filamentous structure of Ebola virions and may suggest an interaction between VP40 and GP in morphogenesis.

Polymorphisms and the differential antiviral activity of the chicken Mx gene.
Ko JH, Jin HK, Asano A, Takada A, Ninomiya A, Kida H, Hokiyama H, Ohara M, Tsuzuki M, Nishibori M, Mizutani M, Watanabe T
Genome research, 12, 4, 595, 601, 4, 2002年04月, ［査読有り］

MLL gene rearrangement in acute myelogenous leukemia after exposure to tegafur/uracil
Toshihiro Fukushima, Nobuyuki Yoshio, Yutaka Noto, Hiroshi Kida
International Journal of Hematology, 75, 2, 178, 181, 2002年02月, ［査読有り］
英語, 研究論文（学術雑誌）, We report a case of acute myelogenous leukemia (AML) with MLL (myeloid-lymphoid leukemia or mixed-lineage leukemia) gene rearrangement after exposure to tegafur/uracil. Cytogenetic and clinical findings in this patient: t(11
17) (q23
q25), AML-M4 morphology, development of AML within a short latent period after first exposure to tegafur/uracil, and good response to remission induction chemotherapy but short remission duration, have been considered typical features of therapy-related acute myelogenous leukemia (t-AML) after exposure to topoisomerase II-targeting agents. This case report suggests that t-AML may develop after exposure to tegafur/uracil and that MLL gene rearrangement may not necessarily be specific to t-AML after exposure to topoisomerase H-targeting agents. ©2002 The Japanese Society of Hematology.

Immunogenicity and protective efficacy of replication-incompetent influenza virus-like particles
T Watanabe, S Watanabe, G Neumann, H Kida, Y Kawaoka
JOURNAL OF VIROLOGY, 76, 2, 767, 773, AMER SOC MICROBIOLOGY, 2002年01月, ［査読有り］
英語, 研究論文（学術雑誌）, Despite the success of influenza virus vaccines in reducing severe illness, their efficacy is suboptimal. We describe here the immunogenicity and protective capacity of replication-incompetent influenza virus-like particles (VLPs) which were generated entirely from cDNAs and lacked either the entire NS gene (encoding both the NS1 and NS2 protein) or only the NS2 gene. In mammalian cells infected with NS gene-deficient VLPs, the nucleoprotein, but not other viral proteins including hemagglutinin (RA) and neuraminidase (NA), was detected. In contrast, cells infected with VLPs expressing NS1 but not NS2 (NS2 knockout) expressed multiple viral proteins, including RA and NA. When challenged with lethal doses of an antigenically homologous mouse-adapted influenza virus, 94% of mice-vaccinated with the NS2-knockout VLPs survived, compared with less than 10% of those given the NS-deficient VLPs. These results demonstrate the potential of replication-incompetent NS2-knockout VLPs as novel influenza vaccines and perhaps also as vectors to express genes from entirely unrelated pathogens.

Serological evidence of transmission of human influenza a and B viruses to Caspian seals (Phoca caspica)
K Ohishi, A Ninomiya, H Kida, CH Park, T Maruyama, T Arai, E Katsumata, T Tobayama, AN Boltunov, LS Khuraskin, N Miyazaki
MICROBIOLOGY AND IMMUNOLOGY, 46, 9, 639, 644, CENTER ACADEMIC PUBL JAPAN, 2002年, ［査読有り］
英語, 研究論文（学術雑誌）, Seroepidemiological surveillance of influenza in Caspian seals (Phoca caspica) was conducted. Antibodies to influenza A virus were detected in 54% (7/13), 57% (4/7), 40% (6/15) and 26% (11/42) of the serum samples collected in 1993, 1997, 1998 and 2000 by enzyme-linked immunosorbent assay (ELISA). In an hemagglutination-inhibition (HI) test using H1-H15 reference influenza A viruses as antigens, more than half of the examined ELISA-positive sera reacted with an H3N2 prototype strain A/Aichi/2/68. These sera were then examined by HI test with a series of naturally occurring antigenic variants of human H3N2 virus, and H3 viruses of swine, duck, and equine origin. The sera reacted strongly with the A/Bangkok/1/79 (H3N2) strain, which was prevalent in humans in 1979-1981. The present results indicate that human A/Bangkok/1/79-like virus was transmitted to Caspian seals probably in the early 1980s, and was circulated in the population. Antibodies to influenza B virus were detected by ELISA in 14% (1/7) and 10% (4/42) serum samples collected from Caspian seals in 1997 and 2000, respectively. Our findings indicate that seal might be a reservoir of both influenza A and B viruses originated from humans.

p38 MAPK phosphorylation and NF-κB activation in human crescentic glomerulonephritis
Norihiko Sakai, Takashi Wada, Kengo Furuichi, Yasunori Iwata, Keiich Yoshimoto, Kiyoki Kitagawa, Satoshi Kokubo, Motoo Kobayashi, Shin-Ichi Takeda, Hiroshi Kida, Ken-Ichi Kobayashi, Naofumi Mukaida, Kouji Matsushima, Hitoshi Yokoyama
Nephrology Dialysis Transplantation, 17, 6, 998, 1004, 2002年, ［査読有り］
英語, 研究論文（学術雑誌）, Background. p38 mitogen-activated protein kinase (p38 MAPK) followed by the activation of NF-κB participates in the intracellular signal transduction and production of cytokines and chemokines. The pathophysiological roles of p38 MAPK and NF-κB in human glomerulonephritis, however, remain to be investigated. Methods. We investigated the phosphorylated p38 MAPK (p-p38 MAPK)and activated NF-κB immunohistochemically in the kidneys of 34 patients with crescentic glomerulonephritis and 26 control patients with thin basement membrane disease and minimal change nephrotic syndrome. We also explored the co-localization of p-p38 MAPK with CCR5, the signal of which leads to p38 MAPK activation. Furthermore, urinary levels of MIP-1α, the cognate ligand for CCR5, were determined by enzyme-linked immunosorbent assay. Results. p-p38 MAPK-positive cells and activated NF-κB-positive cells were mainly detected in crescentic lesions, tubular epithelial cells, and interstitial mononuclear infiltrates. The number of p-p38 MAPK-positive cells in patients with crescentic glomerulonephritis was higher than that in control patients. The number of p-p38 MAPK-positive cells in glomeruli was well correlated with the percentage of cellular crescents, the number of CD68-positive cells, and urinary MIP-1α levels. In addition, the number of activated NF-κB-positive cells was well correlated with the number of p-p38 MAPK-positive cells in glomeruli. Dual staining revealed that most of CCR5-positive cells were positive for p-p38 MAPK. Finally, p-p38 MAPK-positive cells and activated NF-κB-positive cells decreased during glucocorticoid therapy-induced convalescence. Conclusions. We conclude that the phosphorylation of p38 MAPK associated with the activation of NF-κB may be involved in the upregulation of intrarenal MIP-1α and the utilization of CCR5 signalling, which may result in human crescentic glomerulonephritis.

新しいインフルエンザワクチンの開発 "半生"ワクチンのレシピ　　　　　　　　　　　　　　　
渡辺 登喜子, 喜田 宏, 河岡 義裕
感染・炎症・免疫, 31, 4, 318, 320, 鳥居薬品(株), 2001年12月, ［査読有り］
日本語

A serological survey of minute virus of canines (MVC; Canine parvovirus type-1) in dogs in the Tokai area of Japan
A Hashimoto, M Takiguchi, K Hirai, H Kida, LE Carmichael
JAPANESE JOURNAL OF VETERINARY RESEARCH, 49, 3, 249, 253, HOKKAIDO UNIV, 2001年11月, ［査読有り］
英語, 研究論文（学術雑誌）, A serological survey for antibodies to minute virus of canines (WC) by use of a hemagglutination-inhibition (HI) test was performed on sera collected from dogs in the Tokai area of Japan. Forty-one of 266 (15.4%) sera had positive titers of 1 : 40 or higher against the MVC. Results suggest that WC may have been present in dogs in Japan since, at least, 1990. From this serosurvey, WC appears to be established in the dog population in Japan. MVC may have a role as a newly recognized viral pathogen of dogs in Japan.

Characterization of a human H9N2 influenza virus isolated in Hong Kong
T Saito, W Lim, T Suzuki, Y Suzuki, H Kida, SI Nishimura, M Tashiro
VACCINE, 20, 1-2, 125, 133, ELSEVIER SCI LTD, 2001年10月, ［査読有り］
英語, 研究論文（学術雑誌）, Two H9N2 viruses were isolated, for the first time, from humans in Hong Kong in 1999. Isolation Of influenza viruses with a novel subtype of the hemagglutinin (HA) drew attention of health care authorities worldwide from the view of pandemic preparedness. Sequence analysis of the HA genes reveals that HA of A/Hong Kong/1073/99 (H9N2) is most closely related to that of A/quail/HK/G1/97 (H9N2) that contains the internal genes similar to those of Hong Kong/97 (H5N1) viruses. Phylogenetic and antigenic analyses demonstrated the diversity among H9 HA. A/Hong Kong/1073/99 was shown to cause a respiratory infection in Syrian hamsters, suggesting that the virus can replicate efficiently in mammalian hosts. We developed a whole virion test vaccine with a formalin-inactivated egg-grown HK1073. Intraperitoneal administration of the vaccine twice to hamsters conferred a complete protection against challenge infection by the MDCK cell-grown homologous virus. Receptor specificity of HK1073 appeared different from that of other avian influenza viruses of H9 subtype which recognize preferentially alpha -2,3 linked sialic acid. Hemagglutination of HK1073 with guinea pig erythrocytes was inhibited by both alpha -2,3 and alpha -2,6 linked sialic acid containing polymers. These data suggested that HK1073 had acquired a broader host range, including humans. Together with data so far available, the present study suggested that isolation of the H9 influenza viruses from humans requires precaution against the emergence of a novel human influenza. (C) 2001 Elsevier Science Ltd. All rights reserved.

Detection of antibodies to the nonstructural protein (NS1) of influenza A virus allows distinction between vaccinated and infected horses
H Ozaki, T Sugiura, S Sugita, H Imagawa, H Kida
VETERINARY MICROBIOLOGY, 82, 2, 111, 119, ELSEVIER SCIENCE BV, 2001年09月, ［査読有り］
英語, 研究論文（学術雑誌）, Antibodies to the nonstructural protein (NS1) of A/equine/Miami/1/63 (H3N8) influenza virus were detected exclusively in the sera of mice experimentally infected with A/Aichi/2/68 (H3N2) and horses infected with A/equine/Kentucky/1/81 (H3N8) or A/equine/La Plata/1/93 (H3N8), but not in those of the animals immunized with the inactivated viruses, by enzyme-linked immunosorbent assay (ELISA) using a recombinant NS1 as antigen. The results indicate that the present method is useful for serological diagnosis to distinguish horses infected with equine H3 influenza viruses from those immunized with the inactivated vaccine. (C) 2001 Elsevier Science B.V. All rights reserved.

Influenza A virus can undergo multiple cycles of replication without M2 ion channel activity.
Watanabe T, Watanabe S, Ito H, Kida H, Kawaoka Y
Journal of virology, 75, 12, 5656, 5662, 2001年06月, ［査読有り］

Infectivity-Enhancing Antibodies to Ebola Virus Glycoprotein
Takada Ayato, Watanabe Shinji, Okazaki Katsunori, Kida Hiroshi, Kawaoka Yoshihiro
Journal of Virology, 75, 5, 2324, 2330, American Society for Microbiology, 2001年03月
英語, Ebola virus causes severe hemorrhagic fever in primates, resulting in mortality rates of up to 100%, yet there are no satisfactory biologic explanations for this extreme virulence. Here we show that antisera produced by DNA immunization with a plasmid encoding the surface glycoprotein (GP) of the Zaire strain of Ebola virus enhances the infectivity of vesicular stomatitis virus pseudotyped with the GP. Substantially weaker enhancement was observed with antiserum to the GP of the Reston strain, which is much less pathogenic in humans than the Ebola Zaire and Sudan viruses. The enhancing activity was abolished by heat but was increased in the presence of complement system inhibitors, suggesting that heat-labile factors other than the complement system are required for this effect. We also generated an anti-Zaire GP monoclonal antibody that enhanced viral infectivity and another that neutralized it, indicating the presence of distinct epitopes for these properties. Our findings suggest that antibody-dependent enhancement of infectivity may account for the extreme virulence of the virus. They also raise issues about the development of Ebola virus vaccines and the use of passive prophylaxis or therapy with Ebola virus GP antibodies.

Antigenic variation among equine H3N8 influenza virus hemagglutinins
H Ozaki, A Shimizu-Nei, S Sugita, T Sugiura, H Imagawa, H Kida
JAPANESE JOURNAL OF VETERINARY RESEARCH, 48, 4, 177, 186, HOKKAIDO UNIV, 2001年02月, ［査読有り］
英語, 研究論文（学術雑誌）, To provide information on the antigenic variation of the hemagglutinins (HA) among equine H 3 influenza viruses, 26 strains isolated from horses in different areas in the world during the 1963 - 1996 period were analyzed using a panel of monoclonal antibodies recognizing at least 7 distinct epitopes on the H 3 HA molecule of the prototype strain A/equine/Miami/ 1 /63 (H 3 N 8). The reactivity patterns of the virus strains with the panel indicate that antigenic drift of the HA has occurred with the year of isolation, but less extensively than that of human H 3 N 2 influenza virus isolates, and different antigenic variants co-circulate. To assess immunogenicity of the viruses, antisera from mice vaccinated with each of the 7 representative inactivated viruses were examined by neutralization and hemagglutination-inhibition tests. These results emphasize the importance of monitoring the antigenic drift in equine influenza virus strains and to introduce current isolates into vaccine. On the basis of the present results, equine influenza vaccine strain A/equine/Tokyo/ 2 /71 (H 3 N 8) was replaced with A/equine/La Plata/ 1 /93 (H 3 N 8) in 1996 in Japan. The present results of the antigenic analysis of the 26 strains supported the results of a phylogenetic analysis(5)), that viruses belonging to each of the Eurasian and American equine influenza lineages have independently evolved. However, the current vaccine in Japan consists of two American H 3 N 8 strains; A/equine/Kentucky/ 1 /81 and A/equine/La Plata/ 1 /93. It is also therefore recommended that a representative Eurasian strain should be included as a replacement of A/equine/Kentucky/ 1/81.

A rapid and highly sensitive method for diagnosis of equine influenza by antigen detection using immuno-PCR
H Ozaki, S Sugita, H Kida
JAPANESE JOURNAL OF VETERINARY RESEARCH, 48, 4, 187, 195, HOKKAIDO UNIV, 2001年02月, ［査読有り］
英語, 研究論文（学術雑誌）, A rapid and highly sensitive method for diagnosis of influenza by detecting viral antigens using immuno-PCR has been developed. The sensitivity of the immuno-PCR for the detection of the nonstructural protein 1 (NS 1) antigenically common among influenza A viruses was 10(1.0 similar to2.0) times higher than that of RT-PCR for the detection of the viral genome. For the detection of the hemagglutinin (HA) subtype-specific antigen, this assay using anti-HA monoclonal antibodies attained a sensitivity of up to 10(7.0 similar to8.0) times higher than those by virus isolation or by RT-PCR.

Serum cystatin C may predict the prognostic stages of patients with IgA nephropathy prior to renal biopsy
Yasuhiko Tomino, Shigenobu Suzuki, Tomohito Gohda, Michimasa Kobayashi, Satoshi Horikoshi, Hirokazu Imai, Takao Saito, Tetsuya Kawamura, Noriaki Yorioka, Takashi Harada, Yuichiro Yasumoto, Hiroshi Kida, Yutaka Kobayashi, Masayuki Endoh, Hidetoshi Sato, Kensuke Saito
Journal of Clinical Laboratory Analysis, 15, 1, 25, 29, 2001年, ［査読有り］
英語, 研究論文（学術雑誌）, The relationship between the levels of serum cystatin C and the prognostic stages of IgA nephropathy was determined in a multicenter trial in Japan. The levels of serum cystatin C in patients with IgA nephropathy were measured using the Dade Behring N Latex Cystain C assay. In 1995, the Joint Committee of the Special Study Group on Progressive Glomerular Diseases, Ministry of Health and Welfare of Japan, and the Japanese Society of Nephropathy reported four prognostic stages. These are: good prognosis group (Group I), relatively good prognosis group (Group II), relatively poor prognosis group (Group III), and poor prognosis group (Group IV), for this disease. Three-hundred and six patients with IgA nephropathy and other glomerular diseases were examined. There were no significant changes in the levels of serum creatinine (Cr) or creatinine clearance (CCr) between Group I and Group II. The mean levels of serum cystatin C in Group II were significantly higher than those in Group I (P <
0.05). The mean levels of serum cystatin C in Group III or IV were significantly higher than those in Group I (P <
0.001, P <
0.005, respectively). These suggest that the measurement of serum cystatin C may predict the prognostic stages of patients with IgA nephropathy prior to renal biopsy. © 2001 Wiley-Liss, Inc.

Global surveillance of animal influenza for the control of future pandemics
H Kida, K Okazaki, A Takada, H Ozaki, M Tashiro, DK Lvov, KF Shortridge, RG Webster
OPTIONS FOR THE CONTROL OF INFLUENZA IV, 1219, 169, 171, ELSEVIER SCIENCE BV, 2001年, ［査読有り］
英語, 研究論文（国際会議プロシーディングス）, Influenza viruses of different subtypes were isolated from fecal samples of ducks in their nesting areas in Siberia in summer from 1996 to 1999. Phylogenetic analysis of the NP genes of the isolates in Siberia and Japan on their flyway of migration to the south indicate that influenza viruses perpetuated in ducks nesting in Siberia should have contributed genes in the emergence of the H5N1 virus in Hong Kong. Inactivated vaccine prepared from an avirulent H5N4 virus isolated from a migratory duck in Japan showed enough potency to induce protective immunity against the pathogenic H5N1 virus in mice. To expand surveillance for influenza viruses in migratory and domestic ducks and geese, chickens, quail, and pigs of the world and be informed about what influenza viruses are dominant in the animal reservoirs, we have started the "Programme of Excellence in Influenza". Influenza virus strains isolated from animal species in the surveillance should be thoroughly characterized, stored, and be provided for the use of diagnosis and vaccine preparation for the control of future pandemics. (C) 2001 Elsevier Science B.V. All rights reserved.

An efficient plasmid-driven system for the generation of influenza virus-like particles for vaccine
T Watanabe, S Watanabe, G Neumann, H Kida, Y Kawaoka
OPTIONS FOR THE CONTROL OF INFLUENZA IV, 1219, 1015, 1017, ELSEVIER SCIENCE BV, 2001年, ［査読有り］
英語, 研究論文（国際会議プロシーディングス）, We established an efficient plasmid-driven system for the generation of infectious influenza VLPs containing a virus-like RNA segment entirely from cDNAs. To generate influenza VLPs, we used the RNA polymerase I system for the intracellular synthesis of influenza virus RNAs. Human embryonic kidney cells (293T) were transfected with plasmids encoding the influenza A virus structural proteins and with a plasmid encoding an influenza virus-like viral RNA (vRNA), which contained an antisense copy of the cDNA for green fluorescence protein (GFP) flanked by an RNA polymerase I promoter and terminator (pPolI-GFP). Influenza virus-like particles containing GFP vRNA that were infectious and expressed GFP in infected cells were generated. We also generated VLPs lacking the NS gene by eliminating the plasmid for the NS RNA segment from the set of plasmids required for infectious influenza virus production. Because of its efficiency, this system would be useful in studies of influenza virus replication and particle formation as well as for production of vaccines. (C) 2001 Elsevier Science B.V. All rights reserved.

Mucosal vaccination against influenza: protection of pigs immunized with inactivated virus and ether-split vaccine.
Lim YK, Takada A, Tanizaki T, Ozaki H, Okazaki K, Kida H
Japanese Journal of Veterinary Research, 48, 197, 203, 2001年, ［査読有り］
英語, 研究論文（学術雑誌）

Primary target cells of virulent strains of type A influenza virus in chicken embryos.
Park CH, Ozaki H, Takada A, Kida H, Ochiai K, Umemura T
Avian Pathology, 30, 269, 272, 2001年, ［査読有り］
英語, 研究論文（学術雑誌）

Sialyl sugar chains as receptors and determinants of host range of influenza A viruses.　　　　　　　　　　　　　　　
Suzuki Y, Ito T, Suzuki T, Miyamoto D, Kazuya I.-P, J. Hidari, Guo C.-T, Kida H, Webster RG, Chambers TM, Kawaoka Y
Options for the control of influenza IV, 521, 525, 2001年, ［査読有り］
英語, 研究論文（学術雑誌）

Functional Importance of the Coiled-Coil of the Ebola Virus Glycoprotein
Watanabe Shinji, Takada Ayato, Watanabe Tokiko, Ito Hiroshi, Kida Hiroshi, Kawaoka Yoshihiro
Journal of Virology, 74, 21, 10194, 10201, American Society for Microbiology, 2000年11月
英語, Ebola virus contains a single glycoprotein (GP) that is responsible for receptor binding and membrane fusion and is proteolytically cleaved into disulfide-linked GP1 and GP2 subunits. The GP2 subunit possesses a coiled-coil motif, which plays an important role in the oligomerization and fusion activity of other viral GPs. To determine the functional significance of the coiled-coil motif of GP2, we examined the effects of peptides corresponding to the coiled-coil motif of GP2 on the infectivity of a mutant vesicular stomatitis virus (lacking the receptor-binding/fusion protein) pseudotyped with the Ebola virus GP. A peptide corresponding to the C-terminal helix reduced the infectivity of the pseudotyped virus. We next introduced alanine substitutions into hydrophobic residues in the coiled-coil motif to identify residues important for GP function. None of the substitutions affected GP oligomerization, but some mutations, two in the N-terminal helix and all in the C-terminal helix, reduced the ability of GP to confer infectivity to the mutant vesicular stomatitis virus without affecting the transport of GP to the cell surface, its incorporation into virions, and the production of virus particles. These results indicate that the coiled-coil motif of GP2 plays an important role in facilitating the entry of Ebola virus into host cells and that peptides corresponding to this region could act as efficient antiviral agents.

Isolation of Pseudomonas aeruginosa from Ushubetsu River water in Hokkaido, Japan
Yoh Aoi, Hidehiko Nakata, Hiroshi Kida
Japanese Journal of Veterinary Research, 48, 1, 29, 34, 2000年05月, ［査読有り］
英語, 研究論文（学術雑誌）, To provide information on the ecology of Pseudomonas aeruginosa in nature, bacteriological surveillance was performed in the defined area in Hokkaido, Japan. P. aeruginosa was isolated from water samples of Ushubetsu River in the down stream from the urban area of Asahikawa. P. aeruginosa was isolated from fecal samples of pigs but not from samples of soil of a tomato field, sand of sandboxes in vest-pocket parks, fresh vegetables, or feces of wild deer. The present results indicate that P. aeruginosa strains isolated from the river water is originated from the environment of human activity and not from wild life or domestic animals.

Influenza virus-induced encephalopathy: Clinicopathologic study of an autopsied case
Mitsuo Takahashi, Tatsuo Yamada, Yoshio Nakashita, Hiroaki Saikusa, Masatune Deguchi, Hiroshi Kida, Masato Tashiro, Tetsuya Toyoda
Pediatrics International, 42, 2, 204, 214, 2000年, ［査読有り］
英語, 研究論文（学術雑誌）, Background: Rapid progressive encephalopathy with a high fever, consciousness loss and recurrent convulsions has been occasionally reported in children during influenza pandemics in Japan since 1995. We examined a 2- year old girl with hemorrhagic shock and encephalopathy syndrome associated with acute influenza A virus infection (A/Nagasaki/76/98
H3N2), to answer several questions for which no histologic or virologic data exist. Methods: A clinicopathologic study using immunohistochemical staining and viral genome detection by reverse transcriptase polymerase chain reaction (RT-PCR) was performed with this autopsied case. Results: The virus antigen was positive in CD8+ T lymphocytes from the lung and spleen. The virus infected a very limited part of the brain, especially Purkinje cells in the cerebellum and many neurons in the pons, without inducing an overt immunologic reaction from the host. The RT-PCR used for detecting the hemagglutinin gene demonstrated positive bands in all frozen tissues and cerebrospinal fluid taken at autopsy and not in samples obtained on admission. Conclusions: The pathologic change induced by the direct viral invasion cannot be responsible for all of the symptoms, especially for the rapid and severe clinical course of the disease within 24-48 h after the initial respiratory symptoms. Together with the rapid production of several inflammatory cytokines, the breakdown of the blood-brain barrier may induce severe brain edema and can be a major pathologic change for the disease. Any therapeutic strategy to control this multistep progression of the disease could be effective.

Hypothermia with acute renal failure in a patient suffering from diabetic nephropathy and malnutrition
M. Yokoyama, Y. Noto, H. Kida
Diabetes and Metabolism, 26, 2, 145, 147, 2000年, ［査読有り］
英語, 研究論文（学術雑誌）, We report a rare case of hypothermia with acute renal failure in a patient suffering from diabetic nephropathy. A 71-year-old male who had been receiving insulin therapy for the treatment of diabetes mellitus complicated with advanced diabetic nephropathy since 1998 was malnourished with an extremely decreased muscle mass. Without any prolonged exposure to excessively low external temperatures or hypothyroidism, pituitary insufficiency, adrenal insufficiency, sepsis, hypoglycemia, and diabetic ketoacidosis, acute hypothermia appeared together with an aggravation of diabetic nephropathy. His skin temperature fell to below measurable levels and his rectal temperature fell to 30.0°C. His consciousness was drowsy and the hypothermia was not accompanied by shivering. Skeletal muscle is known to play an important role as a center of heat production and shivering thermogenesis in skeletal muscle mainly operates on acute cold stress. Therefore, in this case, hypothermia may have occurred because the shivering thermogenesis could not fully act on the acute cold stress due to the dramatically reduced muscle mass. We should always keep in mind that older, malnourished diabetic patients can easily suffer from impairments of the thermoregulatory system.

Distinct expression of CCR1 and CCR5 in glomerular and interstitial lesions of human glomerular diseases
Kengo Furuichi, Takashi Wada, Norihiko Sakai, Yasunori Iwata, Keiichi Yoshimoto, Miho Shimizu, Ken-Ichi Kobayashi, Kazuya Takasawa, Hiroshi Kida, Shin-Ichi Takeda, Naofumi Mukaida, Kouji Matsushima, Hitoshi Yokoyama
American Journal of Nephrology, 20, 4, 291, 299, S. Karger AG, 2000年, ［査読有り］
英語, 研究論文（学術雑誌）, We investigated the presence of CCR1- and CCR5-positive cells immunohistochemically in the kidneys of 38 patients with several renal diseases, including 13 crescentic glomerulonephritis patients. In addition, we determined cell phenotypes of CCR1- and CCR5-positive cells using a dual immunostaining technique. Urinary levels of their ligands, for CCR1 and CCR5
macrophage inflammatory protein (MIP)-1α, MIP-1β and regulated upon activation in normal T cells expressed and secreted (RANTES) were evaluated by enzyme-linked immunosorbent assay. CCR1- and CCR5-positive cells were detected in both glomeruli and interstitium of the diseased kidneys. Using a dual immunostaining technique, these positive cells were CD68-positive macrophages (MΦ) and CD3-positive T cells. The number of CCR1-positive cells in glomeruli was correlated with urinary levels of MIP-1α. The number of CCR1-positive cells in the interstitium was correlated with both urinary MIP- 1α and RANTES levels. CCR1-positive cells in the interstitium remained after glucocorticoid therapy, most of which were MΦ, and were correlated with the intensity of interstitial fibrosis and tubular atrophy. Glomerular CCR5- positive cells were well correlated with extracapillary lesions and urinary MIP-1α levels, while interstitial CCR5-positive cells, mainly CD3-positive T cells, were correlated with interstitial lesions and urinary RANTES levels. Renal CCR5-positive cells were dramatically decreased during convalescence induced by glucocorticoids. These results suggest that chemokine receptor signaling may be pivotal for human renal diseases through the recruitment and activation of MΦ and T cells
CCR5-positive cells may participate in glomerular lesions including extracapillary lesions via MIP-1α and in interstitial lesions via RANTES. CCR1 may be involved in interstitial lesions in resolving phase after glucocorticoid therapy. Copyright (C) 2000 S. Karger AG, Basel.

Up-regulation of monocyte chemoattractant protein-1 in tubulointerstitial lesions of human diabetic nephropathy
Takashi Wada, Kengo Furuichi, Norihiko Sakai, Yasunori Iwata, Keiichi Yoshimoto, Miho Shimizu, Shin-Ichi Takeda, Kazuya Takasawa, Mitsuhiro Yoshimura, Hiroshi Kida, Ken-Ichi Kobayashi, Naofumi Mukaida, Takero Naito, Kouji Matsushima, Hitoshi Yokoyama
Kidney International, 58, 4, 1492, 1499, 2000年, ［査読有り］
英語, 研究論文（学術雑誌）, Background. We previously described that monocyte chemoattractant protein-1 (MCP-1) plays an important role in progressive glomerular and interstitial damage in inflammatory renal diseases. However, the expression of MCP-1 in diabetic nephropathy remains to be investigated. Methods. We examined whether locally expressed MCP-1 participates in human diabetic nephropathy via recruiting and activating monocytes/macrophages (M∅). Urinary and serum MCP-1 levels were measured by enzyme-linked immunosorbent assay in 45 patients with diabetic nephropathy. The presence of MCP-1 in diseased kidneys was determined by immunohistochemical and in situ hybridization analyses. Results. Urinary MCP-1 levels were significantly elevated in patients with diabetic nephrotic syndrome and advanced tubulointerstitial lesions. Moreover, urinary levels of MCP-1 were well correlated with the number of CD68-positive infiltrating cells in the interstitium. In contrast, serum MCP-1 levels remained similar to those of healthy volunteers. Furthermore, we detected the MCP-1-positive cells in the interstitium of diabetic nephropathy via both immunohistochemical and in situ hybridization analyses. Conclusion. These observations suggest that locally produced MCP-1 may be involved in the development of advanced diabetic nephropathy, especially in the formation of tubulointerstitial lesions possibly through M∅ recruitment and activation. Moreover, up-regulation of MCP-1 may be a common pathway involved in the progressive tubulointerstitial damage in diabetic nephropathy as well as inflammatory renal diseases.

Epitope mapping of the influenza A virus RNA polymerase PA using monoclonal antibodies
M Hatta, Y Asano, K Masunaga, T Ito, K Okazaki, T Toyoda, E Kawaoka, A Ishihama, H Kida
ARCHIVES OF VIROLOGY, 145, 5, 957, 964, SPRINGER-VERLAG WIEN, 2000年, ［査読有り］
英語, 研究論文（学術雑誌）, To obtain reagents to functionally map the PA protein, we produced monoclonal antibodies specific to this protein. Twenty-two monoclonal antibodies reacting with PA protein in ELISA were divided into 10 groups on the basis of competitive binding patterns to this protein. Of these, seventeen monoclonal antibodies bound to PA polypeptide spanning amino acids 101-400 and three bound to that of amino acids 518-600, while the other two did not react with any PA polypeptides tested with the exception of the intact PA. Among these monoclonal antibodies, only five reacted with PA in A/PR/8/34 virus-infected cells in indirect immunofluorescence assay. Thus, we obtained monoclonal antibodies that recognize at least 10 distinct regions of the PA molecule. These monoclonal antibodies should be useful in dissecting functions of the PA protein.

Structure of the infected cell protein 0 gene of canine herpesvirus.
Miyoshi M, Takiguchi M, Yasuda J, Hashimoto A, Takada A, Okazaki K, Kida H
Archives of Virology, 145, 8, 1715, 1723, 2000年, ［査読有り］
英語, 研究論文（学術雑誌）

Downregulation of  1 integrins by Ebola virus glycoprotein: Implication for virus entry.
Takada A, Watanabe S, Ito H, Okazaki K, Kida H, Kawaoka Y
Virology, 278, 1, 20, 26, 2000年, ［査読有り］
英語, 研究論文（学術雑誌）

Mapping of functional domains on the influenza A virus RNA polymerase PB2 molecule using monoclonal antibodies
Hatta M, Asano Y, Masunaga K, Ito T, Okazaki K, Toyoda T, Kawaoka Y, Ishihama A, Kida H
Arch. Virol., 145, 9, 1947, 1961, 2000年, ［査読有り］
英語, 研究論文（学術雑誌）

Avian influenza virus intranasally inoculated infects the central nervous system of mice through the general visceral afferent nerve
Sinya K, Shimada A, Ito T, Umemura T, Tanaka H, Takada A, Kida H, Morita T, Otsuki K
Arch. Virol., 145, 1, 187, 195, 2000年, ［査読有り］
英語, 研究論文（学術雑誌）

Epitope mapping of the influenza A virus RNA polymerase PA using monoclonal antibodies.
Hatta M, Asano Y, Masunaga K, Ito T, Okazaki K, Toyoda T, Kawaoka Y, Ishihama A, Kida H
Arch. Virol., 145, 5, 895, 903, 2000年, ［査読有り］
英語, 研究論文（学術雑誌）

Precursor genes of future pandemic influenza viruses are perpetuated in ducks nesting in Siberia.
Okazaki K, Takada A, Ito T, Imai M, Takakuwa H, Hatta M, Ozaki H, Tanizaki T, Nagano T, Ninomiya A, Demenev VA, Tyaptirganov MM, Yamnikova SS, Lvov DK, Kida H
Arch.Virol., 145, 5, 885, 893, 2000年, ［査読有り］
英語, 研究論文（学術雑誌）

Recognition of N-glycolyneuraminic acid linked to galactose by α2,3 linkage associated with intestinal replication of influenza A virus in ducks
Ito T, Suzuki Y, Suzuki T, Takada A, Horimoto T, Wells K, Kida H, Otsuki K, Kawaoka Y
J. Virol., 74, 19, 9300, 9305, 2000年, ［査読有り］
英語, 研究論文（学術雑誌）

Avirulent Avian Influenza Virus as a Vaccine Strain against a Potential Human Pandemic
Takada Ayato, Kuboki Noritaka, Okazaki Katsunori, Ninomiya Ai, Tanaka Hiroko, Ozaki Hiroichi, Itamura Shigeyuki, Nishimura Hidekazu, Enami Masayoshi, Tashiro Masato, Shortridge Kennedy F, Kida Hiroshi
Journal of Virology, 73, 10, 8303, 8307, American Society for Microbiology, 1999年10月
英語, In the influenza H5N1 virus incident in Hong Kong in 1997, viruses that are closely related to H5N1 viruses initially isolated in a severe outbreak of avian influenza in chickens were isolated from humans, signaling the possibility of an incipient pandemic. However, it was not possible to prepare a vaccine against the virus in the conventional embryonated egg system because of the lethality of the virus for chicken embryos and the high level of biosafety therefore required for vaccine production. Alternative approaches, including an avirulent H5N4 virus isolated from a migratory duck as a surrogate virus, H5N1 virus as a reassortant with avian virus H3N1 and an avirulent recombinant H5N1 virus generated by reverse genetics, have been explored. All vaccines were formalin inactivated. Intraperitoneal immunization of mice with each of vaccines elicited the production of hemagglutination-inhibiting and virus-neutralizing antibodies, while intranasal vaccination without adjuvant induced both mucosal and systemic antibody responses that protected the mice from lethal H5N1 virus challenge. Surveillance of birds and animals, particularly aquatic birds, for viruses to provide vaccine strains, especially surrogate viruses, for a future pandemic is stressed.

Resistance to pseudorabies virus infection in transgenic mice expressing the chimeric transgene that represses the immediate-early gene transcription.
Ono E, Tasaki T, Kobayashi T, Taharaguchi S, Nikami H, Miyoshi I, Kasai N, Arikawa J, Kida H, Shimizu Y
Virology, 262, 1, 72, 78, 1, 1999年09月, ［査読有り］

Replacement of Internal Protein Genes, with the Exception of the Matrix, in Equine 1 Viruses by Equine 2 Influenza Virus Genes during Evolution in Nature
Toshihiro Ito, Yoshihiro Kawaoka, Maki Ohira, Hiroki Takakuwa, Jiro Yasuda, Hiroshi Kida, Koichi Otsuki
Journal of Veterinary Medical Science, 61, 8, 987, 989, 1999年08月, ［査読有り］
英語, 研究論文（学術雑誌）, To establish the evolutionary association between the equine 1 H7 HA and M genes, phylogenetic analyses of the six internal gene segments of equine 1 influenza viruses (H7N7 subtype) were performed using partial nucleotide sequences. The results demonstrated that five internal genes (PB1, PB2, PA, NP and NS) of equine 1 viruses isolated after 1964 were replaced by those of equine 2 H3N8 viruses. However, the M gene was maintained during the evolution of these equine 1 viruses. These findings suggest a functional association between equine H7 HA and M gene products, most likely M2 protein.

Differences in Receptor Specificity between Newcastle Disease Viruses Originating from Chickens and Waterfowl
Toshihiro Ito, Yoshihiro Kawaoka, Chinatsu Kameda, Jiro Yasuda, Hiroshi Kida, Koichi Otsuki
Journal of Veterinary Medical Science, 61, 8, 951, 953, 1999年08月, ［査読有り］
英語, 研究論文（学術雑誌）, We compared the receptor specificity of Newcastle disease viruses from a variety of avian species, including chickens and wild waterfowl, using hemagglutination tests with erythrocytes from different animal species. All isolates from wild waterfowl agglutinated horse erythrocytes, while the chicken isolates did not. The results showed that the receptor specificity of Newcastle disease viruses is different, depending on the avian species from which the viruses are isolated.

Detection of Canine Herpesvirus DNA in the Ganglionic Neurons and the Lymph Node Lymphocytes of Latently Infected Dogs
Masahiro Miyoshi, Yuki Ishii, Mitsuyoshi Takiguchi, Ayato Takada, Jun Yasuda, Akira Hashimoto, Katsunori Okazaki, Hiroshi Kida
Journal of Veterinary Medical Science, 61, 4, 375, 379, 1999年04月, ［査読有り］
英語, 研究論文（学術雑誌）, To determine the site of latent infection of canine herpesvirus (CHV), tissues from dogs convalescent from acute infection with CHV were examined for the presence of viral genome DNA by the nested polymerase chain reaction. CHV DNA was detected in the trigeminal ganglia and the retropharyngeal lymph nodes. In situ hybridization study of the tissues revealed that CHV genome persisted in the nuclei of ganglionic neurons and lymphocytes.

Antigenic and genetic analyses of H5 influenza viruses isolated from ducks in Asia
IMAI Masaki, TAKADA Ayato, OKAZAKI Katsunori, KIDA Hiroshi
Japanese journal of veterinary research, 46, 4, 171, 177, 北海道大学, 1999年02月26日
英語, The hemagglutinin (HA) of six H5 influenza virus strains isolated from ducks in Japan and China in 1976 to 1996 were analyzed antigenically and genetically. Antigenic analysis using a panel of monoclonal antibodies revealed that the HA of H5 influenza viruses isolated from ducks are antigenically closely related to each other. Phylogenetic analysis indicates that the isolates from ducks in Hokkaido were derived from an ancestor common with the highly pathogenic isolates from chickens and humans in Hong Kong in 1997.

The role of monocyte chemotactic and activating factor(MCAF)/monocyte chemoattractant protein (MCP)-l in subgroups of rapidly progressive glomerulonephritis
Norihiko Sakai, Takashi Wada, Kengo Furuichi, Chikako Takaeda, Miho Shimizu, Yasunori Iwata, Kei-Ichi Yoshimoto, Kazuaki Shimizu, Ken-Ichi Kobayashi, Shin-Ichi Takeda, Hiroshi Kida, Naofumi Mukaida, Kouji Matsushima, Hitoshi Yokoyama
Japanese Journal of Nephrology, 41, 7, 704, 711, 1999年, ［査読有り］
英語, 研究論文（学術雑誌）, To elucidate the role of monocyte chemotactic and activating factor(MCAF)/monocyte chemoattractant protein (MCP)-l in the pathogenesis of rapidly progressive glomerulonephritis(RPGN), we determined the urinary levels of MCAF/MCP-1 in 20 healthy subjects, 30 patients showing RPGN with crescents, and 39 patients with various types of renal diseases without crescents. We divided RPGN into two subgroups, the acute type and the insidious type, with regard to the declination rate of reciprocals of serum creatinine with time as previously reported. In addition, we divided the patients with RPGN into anti-neutrophil cytoplasmic antibody (ANCA)-related diseases and immune complex (1C)-mediated diseases with regard to etiology. Urinary levels of MCAF/MCP-1 were significantly higher in patients with RPGN as compared with those of other renal diseases and healthy volunteers (21.8 ±4.5 vs. 11.6±3.5, 1.0±0.1 pg/m/ creatinine, respectively, p<
0.01, mean±SEM). There was no difference in the urinary levels of MCAF/MCP-1 between the acute and insidious types of RPGN patients. In addition, there was no difference in the urinary levels of MCAF/MCP-1 between the patients with ANCA-related and ICmediated diseases. Urinary levels of MCAF/MCP-1 in patients with RPGN were correlated well with the percentage of both total crescents and fibrocellular/fibrous crescents and the number of CD68-positive infiltrating cells in the interstitium. Immunohistochemical examinations revealed that MCAF/MCP-1positive cells were detected in tubular epithelial and endothelial cells and mononuclear infiltrated cells in the interstitium. Moreover, elevated urinary MCAF/MCP-1 levels in patients with RPGN, regardless of subgroups, were dramatically dereased during methylprednisolone pulse therapy-induced convalescence. These results suggest that MCAF/MCP-1 may be involved in the pathogenesis of RPGN via macrophage recruitment and activation.

The short- and long-term outcomes of membranous nephropathy treated with intravenous immune globulin therapy
Hitoshi Yokoyama, Satoshi Goshima, Takashi Wada, Masayoshi Takaeda, Kengo Furuichi, Ken-Ichi Kobayashi, Hiroshi Kida
Nephrology Dialysis Transplantation, 14, 10, 2379, 2386, 1999年, ［査読有り］
英語, 研究論文（学術雑誌）, Background. A considerable diversity in prognosis is seen with membranous nephropathy (MN). A recent report showed beneficial effects of immune globulin (Glb) therapy in Heymann nephritis, a rat model of MN. However, the early and late clinical effects of Glb in human MN have remained unclear. Methods. We studied retrospectively 86 patients with primary MN from 1965 to 1988 who were followed for at least 5 years, or until renal or actual death. Thirty patients were non-randomly treated with 1-3 courses of intravenous immune globulin, 5-10 g/day (100-150 mg/kg/day) for 6 consecutive days. Based on electron microscopic (EM) findings, the patients were divided into two subtypes, i.e. homogeneous type with synchronous electron-dense deposits, and heterogeneous type with various stages of dense deposits, due to their different clinical outcomes. Results. There was no difference in the initial clinicopathological states between Glb (n = 30) and non-Glb group (n = 56) (70 vs 68% in nephrotic state
37 vs 39% in female, 50 vs 52% in homogeneous type, 50 vs 48% in heterogeneous type respectively). For the homogeneous type, at 6 months post-treatment, Glb therapy had induced earlier remission as compared to non-Glb treatments with corticosteroid alone or together with cyclophosphamide (57 vs 10% respectively, P = 0.006). However, there was no significant difference in the early therapeutic effect for the heterogeneous type (13% for Glb vs 5% for non-Glb in remission after 6 months), or in the final outcome for all groups (18% for Glb vs 10% for non-Glb in renal death after 15 years). No adverse effects were recorded during or after Glb therapy. Conclusions. Our results suggest that short-term relatively low-dose intravenous Glb therapy has a beneficial effect in the earlier induction of remission in a subgroup of MN, the homogeneous type with EM findings of synchronous electron-dense deposits, but does not alter the long-term outcome of human MN.

Structure of the immediate early gene of canine herpesvirus.
Miyoshi M, Takiguchi M, Yasuda J, Hashimoto A, Takada A, Okazaki K, Kida H
Archives of Virology, 144, 2, 407, 420, 1999年, ［査読有り］
英語, 研究論文（学術雑誌）

Induction of antibodies in mice by a recombinant baculovirus expressing pseudorabies virus glycoprotein B in mammalian cells.
Aoki H, Sakoda Y, Jukuroki K, Takada A, Kida H, Fukusho A
Veterinary Microbiology, 68, 197, 207, 1999年, ［査読有り］
英語, 研究論文（学術雑誌）

Protective effects of intranasal vaccination with plasmid encoding pseudorabies virus glycoprotein B in mice.　　　　　　　　　　　　　　　
Takada A, Okazaki K, Kida H
Japanese Journal of Veterinary Research, 47, 25, 33, 1999年, ［査読有り］
英語, 研究論文（学術雑誌）

Substitution of amino acid residue in influenza A virus hemagglutinin affects recognition of sialyl-oligosaccharides containing N-glycolylneuraminic acid.
Masuda H, Suzuki T, Sugiyama Y, Horiike G, Murakami K, Miyamoto D, Hidari I-PJK, Ito T, Kida H, Kiso M, Fukunaga K, Ohuchi T, Toyoda T, Ishihama A, Kawaoka Y, Suzuki Y
FEBS Lett., 464, 1月2日, 71, 74, 1999年, ［査読有り］
英語, 研究論文（学術雑誌）

Replacement of internal protein genes, with the exception on the matrix, ix equine 1 viruses by equine 2 influenza virus genes during evolution in nature
Ito T, Kawaoka Y, Ohira M, Takakuwa H, Yasuda J, Kida H, Otsuki K
J. Vet. Med. Sci., 61, 8, 987, 989, 1999年, ［査読有り］
英語, 研究論文（学術雑誌）

Differences in receptor specificity between newcastle disease viruses originating from chickens and waterfowl
Ito T, Kawaoka Y, Kameda C, Yasuda J, Kida H, Otsuki K
J. Vet. Med. Sci, 61, 8, 951, 953, 1999年, ［査読有り］
英語, 研究論文（学術雑誌）

Pseudorabies virus (PRV) early protein 0 activates PRV gene transcription in combination with the immediate-early protein IE180 and enhances the infectivity of PRV genomic DNA.
Ono E, Watanabe S, Nikami H, Tasaki T, Kida H
Veterinary microbiology, 63, 2-4, 99, 107, 1998年10月, ［査読有り］

Molecular basis for the generation in pigs of influenza A viruses with pandemic potential
T Ito, JNSS Couceiro, S Kelm, LG Baum, S Krauss, MR Castrucci, Donatelli, I, H Kida, JC Paulson, RG Webster, Y Kawaoka
JOURNAL OF VIROLOGY, 72, 9, 7367, 7373, AMER SOC MICROBIOLOGY, 1998年09月, ［査読有り］
英語, 研究論文（学術雑誌）, Genetic and biologic observations suggest that pigs may serve as "mixing vessels" for the generation of human-avian influenza A virus reassortants, similar to those responsible for the 1957 and 1968 pandemics. Here we demonstrate a structural basis for this hypothesis. Cell surface receptors for both human and avian influenza viruses were identified in the pig trachea, providing a milieu conducive to viral replication and genetic reassortment. Surprisingly, with continued replication, some avian-like swine viruses acquired the ability to recognize human virus receptors, raising the possibility of their direct transmission to human populations. These findings help to explain the emergence of pandemic influenza viruses and support the need for continued surveillance of swine for viruses carrying avian virus genes.

Promoter activity of sequence located upstream of the pseudorabies virus early protein 0 gene.
Watanabe S, Ono E, Nikami H, Kida H
Veterinary microbiology, 61, 1-2, 7, 19, 1998年03月, ［査読有り］

Suppression of pseudorabies virus replication by a mutant form of immediate-early protein IE180 repressing the viral gene transcription.
Ono E, Taharaguchi S, Watanabe S, Nikami H, Shimizu Y, Kida H
Veterinary microbiology, 60, 2-4, 107, 117, 1998年02月, ［査読有り］

Fusion of influenza virus with the endosomal membrane is inhibited by monoclonal antibodies to defined epitopes on the hemagglutinin.
Imai M, Sugimoto K, Okazaki K, Kida H
Virus research, 53, 2, 129, 139, 2, 1998年02月, ［査読有り］

Characteristics of Renal Survival Rate in Crescentic Glomerulonephritis -Analysis Using Weibull'S Distribution Function
Hiroshi Kida, Mitsuhiro Yoshimura, Kazuya Takasawa, Tamayo Kato, Kishichiro Watanabe, Keiko Kawabata
IRYO - Japanese Journal of National Medical Services, 52, 10, 596, 600, 1998年, ［査読有り］
英語, 研究論文（学術雑誌）, In a present situation of “try and error” on treating crescentic glomerulonephritis, it is essential to firmly grasp an actual mode of rapid decrease in renal survival rate observed in its early phase. Thus, Weibull's distribution function was applied for analysis of cumulative renal death rate, being calculated as 1-renal survival rate. Data analysed were obtained from 75 patients, in whom renal histological examinations revealed crescent formation in more than 50% of glomeruli observed, and analyses provided following results : l) Cumulative renal death rate showed biphasic rectilinear regression, more steep in the early and less steep in the late phase on Weibull's chart and its shifting time-point was calculated to be 8.8 months after a calculated date of disease onset. 2) The steep increase depended mainly on individual death rate. 3) Similar steep increase was observed in a pauci-immune type but not in an immune complex type and in an acute type but not in an insidious type. Based on these results, those patients presenting the acute type superimposed on the pauci-immune type were marked off as a high risk group. © 1998, Japanese Society of National Medical Services. All rights reserved.

Methylprednisolone pulse therapy in two clinical types of crescentic glomerulonephritis
Shin-Ichi Takeda, Hiroshi Kida, Hitoshi Yokoyama, Eisuke Takazakura, Ken-Ichi Kobayashi
Internal Medicine, 37, 7, 585, 591, Japanese Society of Internal Medicine, 1998年, ［査読有り］
英語, 研究論文（学術雑誌）, To clarify the effectiveness of methylprednisolone pulse therapy on crescentic glomerulonephritis (CresGN), data of 46 patients was retrospectively studied. According to the slope of reciprocals of serum creatinine (s-Cr) the patients were divided into acute and insidious types, and each type was further divided into pulse (P) and conventional (C) therapy groups. In group C, s-Cr improved more frequently in the acute type (8/10) than in the insidious type (1/9, p<
0.05), and the renal survival rate was somewhat higher in the former (p=0.09). In the acute type there was no difference in the improvement rate of s-Cr between the two therapy groups, whereas in the insidious type, the improvement rate was higher in group P (9/15) than in group C (1/9, p<
0.05) and the renal survival rate was higher in the former (p<
0.01). These results suggest that methylprednisolone pulse therapy may be highly effective for the insidious type of CresGN.

Molecular basis for the generation of pandemic influenza viruses in pigs.　　　　　　　　　　　　　　　
Ito T, Nelson J, Couseiro SS, Kelm S, Baum LG, Castrucci MR, Donatelli I, Kida H, Paulson JC, Webster RG, Kawaoka Y
J. Virol., 72, 9, 7367, 7373, 1998年, ［査読有り］
英語, 研究論文（学術雑誌）

An attenuation mechanism of Newcastle disease vaccine strain TCND.
Takakuwa H, Ito T, Takada A, Okazaki K, Kida H
Arch. Virol, 143, 6, 1129, 1143, 1998年, ［査読有り］
英語, 研究論文（学術雑誌）

Continued circulation of reassortant H1N2 influenza viruses in pigs in Japan.
Ito T, Kawaoka Y, Vines A, Ishikawa H, Asai T, Kida H
Arch. Virol., 143, 9, 1773, 1782, 1998年, ［査読有り］
英語, 研究論文（学術雑誌）

Potential virulent Newcastle disease viruses are maintained in migratory waterfowl populations.
Takakuwa H, Ito T, Takada A, Okazaki K, Kida H
Jap. J. Vet. Res., 45, 4, 207, 215, 1998年, ［査読有り］
英語, 研究論文（学術雑誌）

Effects of methylprednisolone pulse therapy on the insidious type of crescentic glomerulonephritis
Shin-Ichi Takeda, Hiroshi Kida, Eisuke Takazakura, Hitoshi Yokoyama, Ken-Ichi Kobayashi
Japanese Journal of Nephrology, 39, 5, 490, 496, 1997年, ［査読有り］
日本語, 研究論文（学術雑誌）, In an attempt to clarify the effects of methylprednisolone pulse therapy on the insidious (subacute) type of crescentic glomerulonephritis with slow, but steady deterioration of renal function and poor response to treatment, we analyzed the clinical course of 24 patients (male : female=15 : 9) with a mean age of 48.5 years. They fulfilled the following criteria : 1) crescents were observed in more than 50% of the glomeruli, 2) the increment of serum creatinine (Cr) could be determined sequentially on three or more occasions before treatment, and reciprocals of serum Cr declined with slopes of less than 1.0 x 10-2 dl/mg/day, 3) corticosteroids and/or immunosuppressants were administered. The patients were divided into two groups : pulse therapy group (P) (15 patients), to which methylprednisolone 500 or 1,000 mg a day was administered intravenously for three consecutive days, and a conventional therapy group (C) (9 patients). There were no differences between groups P and C in clinical parameters, including sex, age, underlying diseases, urinary protein, blood pressure, serum Cr and slope of 1/Cr before treatment, and pathological findings, including percentages of glomeruli with crescents and degree of interstitial lesion. However, improvement of serum Cr, which was defined as a decline to the normal range or less than half of the pretreatment level, was observed in 9 (60%) in group P vs. only 1 (11%) in group C (p<
0.05). Re-biopsies were performed after treatment in 6 patients of group P with an improvement of serum Cr, and showed a decrease in the rate of crescent formation and almost complete loss of cellular crescents. A 1, 2 and 3 years of follow-up, the renal survival rates were 86, 70 and 53%, respectively, in group P vs. 67, 14 and 14% respectively, in group C (p<
0.05). No serious side effects were observed in group P. These results suggest that methylprednisolone pulse therapy may be very effective for the insidious type of crescentic glomerulonephritis.

Spinal cord compression by heterotopic ossification associated with pseudohypoparathyroidism
Y. Yamamoto, Y. Noto, M. Saito, H. Ichizen, H. Kida
Journal of International Medical Research, 25, 6, 364, 368, SAGE Publications Ltd, 1997年, ［査読有り］
英語, 研究論文（学術雑誌）, This report describes a 37-year-old man presenting with a gait disturbance due to spastic paraparesis. Physical findings showed typical features of Albright's hereditary osteodystrophy, including short stature, obesity, brachydactyly and dental hypoplasia. He was diagnosed as having pseudohypoparathyroidism type Ia, on the basis of his hypocalcaemia, hyperphosphataemia, increased plasma level of parathyroid hormone (PTH), and the unresponsiveness to exogenous PTH loading of his urinary excretion of both nephrogenous cyclic adenosine monophosphate and phosphate. Magnetic resonance imaging and myelographic computed tomographic scans clearly demonstrated severe compression of the spinal cord at T 9/10 by tumour-like ossifications of the paravertebral ligaments. Neurosurgical decompression therapy was, therefore, performed to alleviate his spastic paraparesis. This was a rare case of pseudohypoparathyroidism complicated with spinal cord compression caused by ectopic ossification of the ligaments.

Adaptation of equine herpesvirus 1 to unnatural host led to mutation of the gC resulting in increased susceptibility of the virus to heparin.
Sugahara Y, Matsumura T, Kono Y, Honda E, Kida H, Okazaki K
Archives of virology, 142, 9, 1849, 1856, 9, 1997年, ［査読有り］

Swine influenza virus strains recognize sialylsugar chains containing the molecular species of sialic acid predominantly present in the swine tracheal epitherium.
Suzuki T, Horiike G, Yamazaki Y, Kawabe K, Masuda H, Miyamoto D, Matsuda M, Nishimura S, Yamagata T, Ito T, Kida H, Kawaoka Y, Suzuki Y
FEBS Letter, 404, 2月3日, 192, 196, 1997年, ［査読有り］
英語, 研究論文（学術雑誌）

Differences in sialic acid-galactose linkages in the amnion and allantois of chicken eggs influence human influenza virus receptor specificity and variant selection.　　　　　　　　　　　　　　　
Ito T, Suzuki Y, Takada A, Kawamoto A, Otsuki K, Masuda H, Yamada M, Suzuki T, Kida H, Kawaoka Y
Journal of Virology, 71, 3357, 3362, 1997年, ［査読有り］
英語, 研究論文（学術雑誌）

Receptor specificity of influenza A viruses correlates with the agglutination of erythrocytes from different animal species.
Ito T, Suzuki Y, Mitnaul L, Vines A, Kida H, Kawaoka Y
Virology, 227, 2, 493, 499, 1997年, ［査読有り］
英語, 研究論文（学術雑誌）

Bovine clostridial infections in Zambia (1985-1994)
HM Munangandu, PM Muyoyeta, AS Mweene, H Kida
JAPANESE JOURNAL OF VETERINARY RESEARCH, 44, 3, 175, 178, HOKKAIDO UNIV, 1996年12月, ［査読有り］
英語, 研究論文（学術雑誌）, Retrospective surveillance study of clostridial infections of cattle in Zambia, for the period 1985 to 1994, showed that out of the 318 cases observed, 62.8% and 24.2% were from Western and Southern provinces, respectively. Of the 6 clostridia specie's identified, Clostridium septicum (38.1%) followed by C. chauvoei (36.2%) and C. perfringens (13.2%) were dominant. Although the highest incidence for clostridial infections was in 1989 (75 cases) and 1990 (77 cases), the number of C. perfringens cases seemed to increase. More cases were found in the dry season until the onset of the rains, that is, the period August to December.

Negative regulation of immediate-early gene expression of pseudorabies virus by interferon-alpha
N Tonomura, E Ono, Y Shimizu, H Kida
VETERINARY MICROBIOLOGY, 53, 3-4, 271, 281, ELSEVIER SCIENCE BV, 1996年12月, ［査読有り］
英語, 研究論文（学術雑誌）, Pseudorabies virus (PrV) replication in Vero cells was suppressed by treatment with human natural interferon-alpha (IFN-alpha). Messenger RNA transcribed from the PrV immediate-early (IE) gene was reduced in the IFN-alpha-treated cells, Transient expression assays showed that transcription from the PrV IE promoter was selectively inhibited in the IFN-alpha-treated cells. Analysis of deletion mutants of the PrV LE promoter sequence suggested that at least one element between the transcription initiation site (+1) and -90 in the PrV IE promoter was concerned with the negative regulation.

Detection of viral genome in non-neural tissues of cattle experimentally infected with bovine herpesvirus 1
AS Mweene, K Okazaki, H Kida
JAPANESE JOURNAL OF VETERINARY RESEARCH, 44, 3, 165, 174, HOKKAIDO UNIV, 1996年12月, ［査読有り］
英語, 研究論文（学術雑誌）, To obtain further information on the latent infection with bovine herpesvirus 1 (BHV-1) in the natural host, we examined the nasal secretions and various tissues of experimentally infected calves using virus isolation, polymerase chain reaction (PCR), and immuno-PCR/antigen (Immuno-PCR/Ag) assays. In the nasal secretions, viral DNA was detected in samples with virus isolation titers of 10(4.3) TCID50 or more by PCR. On the same samples, Immuno-PCR/Ag remained positive up to day 19 p.i. the last day of test. BHV-1 DNA was detected from the following tissues in all the calves at day 22 p.i.: trigeminal ganglia, ovaries, lungs, nasal and tracheal mucosae, spleen, prescapular and precrural lymph nodes, and peripheral blood leukocytes (PBL), but not skeletal muscles. Virus was not recovered from any of these tissues. The present findings suggest a possible role of leukocytes in BHV-1 latent infection.

Survey of the hemagglutinin (HA) cleavage site sequence of H5 and H7 avian influenza viruses: Amino acid sequence at the HA cleavage site as a marker of pathogenicity potential
DA Senne, B Panigrahy, Y Kawaoka, JE Pearson, J Suss, M Lipkind, H Kida, RG Webster
AVIAN DISEASES, 40, 2, 425, 437, AMER ASSOC AVIAN PATHOLOGISTS, 1996年04月, ［査読有り］
英語, 研究論文（学術雑誌）, The deduced amino acid sequence at the hemagglutinin (HA) cleavage site of 76 avian influenza (AI) viruses, subtypes H5 and H7, was determined by reverse transcription-polymerase chain reaction and cycle sequencing techniques to assess pathogenicity Eighteen of the 76 viruses were isolated in 1993 and 1994 from various sources in the United States. In addition, 34 H5 (4 highly pathogenic [HP] and 30 non-highly pathogenic [non-HP]) and 24 H7 (3 HP and 21 non-HP) repository viruses, isolated between 1927 and 1992, were sequenced and the sequences compared to chose in recent isolates. All repository HP H5 and H7 viruses studied had multiple basic amino acids adjacent to the HA cleavage site and most had basic amino acids in excess of the proposed minimum motif B-X-B-R (B = basic amino acids arginine or lysine, X = nonbasic amino acid, R = arginine) chat has been associated with high pathogenicity. Of the non-HP viruses studied, 35 of 38 for H5 and 30 of 31 for H7 conformed to the motif B-X-X-R and B-X-R, respectively. Two non-HP H5 viruses had the mot if X-X-X-R ar the cleavage sire and a third had the motif B-X-X-K (K = basic amino acid lysine). One non-KP H7 (A/Pekin robin/CA/30412-5/94) had four basic amino acids (K-R-R-R) adjacent to the cleavage sire. Although the Pekin robin isolate did nor produce disease in chickens under the conditions of the study it did have che amino acid sequence compatible with that in HP AI viruses and, therefore, is considered potentially HP This is the first account of an H7 virus that is non-HP in chickens but meets the molecular criterion to be classified as HP.

Mapping of transregulatory domains of pseudorabies virus early protein 0 and identification of its dominant-negative mutant
S Watanabe, E Ono, Y Shimizu, H Kida
ARCHIVES OF VIROLOGY, 141, 6, 1001, 1009, SPRINGER-VERLAG WIEN, 1996年, ［査読有り］
英語, 研究論文（学術雑誌）, Pseudorabies virus (PRV) early protein 0 (EP0) is a transactivator containing the RING finger domain. Analysis of transactivating activity of truncated forms of the EP0 molecule consisting of 410 amino acids revealed that amino-terminal region containing the RING finger domain, amino acids 1 to 84, and the region between amino acids 114 to 242 containing acidic amino acid sequences were required for the transactivation. On the other hand, the mutant consisting of amino acids 1 to 113 exhibited a dominant-negative property.

Protective immune response of chickens against Newcastle disease, induced by the intranasal vaccination with inactivated virus.
Takada A, Kida H
Veterinary Microbiology, 50, 17, 25, 1996年, ［査読有り］
英語, 研究論文（学術雑誌）

Avian influenza viruses as the origin of pandemic strains : perpetuation in nature and potential for transmission to pigs.　　　　　　　　　　　　　　　
Kida H, Ito T, Okazaki K, Kawaoka Y, Webster RG
Options for the control of influenza III, 529, 536, 1996年, ［査読有り］
英語, 研究論文（学術雑誌）

Receptors of influenza A viruses : Implications for the role of pigs for the generation of pandemic human influenza viruses .　　　　　　　　　　　　　　　
Ito T, Kida H, Kawaoka Y
Options for the control of influenza III, 516, 519, 1996年, ［査読有り］
英語, 研究論文（学術雑誌）

Development of immuno-PCR for diagnosis of bovine herpes virus 1 infection.　　　　　　　　　　　　　　　
Mweene AS, Ito T, Okazaki K, Ono E, Shimizu Y, Kida, H
Journal of Clinical Microbiology, 34, 748, 750, 1996年, ［査読有り］
英語, 研究論文（学術雑誌）

Pseudorabies virus early protein 0 transactivates the viral gene promoters.
Watanabe S, Ono E, Shimizu Y, Kida H
The Journal of general virology, 76 ( Pt 11), 2881, 2885, 1995年11月, ［査読有り］

Inhibition of pseudorabies virus replication by a chimeric trans-gene product repressing transcription of the immediate-early gene.
Ono E, Sakoda Y, Taharaguchi S, Watanabe S, Tonomura N, Kida H, Shimizu Y
Virology, 210, 1, 128, 140, 1995年06月, ［査読有り］

MAPPING OF A FUNCTIONAL REGION CONFERRING NUCLEAR-LOCALIZATION OF PSEUDORABIES VIRUS IMMEDIATE-EARLY PROTEIN
S TAHARAGUCHI, E ONO, S YAMADA, Y SHIMIZU, H KIDA
ARCHIVES OF VIROLOGY, 140, 10, 1737, 1746, SPRINGER-VERLAG WIEN, 1995年, ［査読有り］
英語, 研究論文（学術雑誌）, The immediate-early protein (IE180) of pseudorabies virus (PrV) is localized predominantly in the nuclei of infected cells. To define the nuclear localization signals within IE180, we prepared truncated mutants of IE180 and analyzed their localization in the transfected cells by indirect immunofluorescence. Analysis of mutants truncated from the carboxy-terminal end of the 1460-amino acid polypeptide showed that two regions including a short sequence of basic amino acid residues were associated with the nuclear localization of IE180. To assess whether these regions substantially function as signals for nuclear localization of the IE180 molecule, we then constructed two deletion mutants lacking each region. A mutant lacking amino acids 333 to 575 was detected in the nuclei of the transfected cells, whereas the other mutant lacking amino acids 900 to 950 was detected mainly in the cytoplasm. These results suggest that the region of amino acids 900 to 950 is responsible for nuclear localization of IE180.

Induction of protective antibody responses against pseudorabies virus by intranasal vaccination with glycoprotein B in mice.
Takada A, Kida H
Archives of Virology, 140, 9, 1629, 1635, 1995年, ［査読有り］
英語, 研究論文（学術雑誌）

Perpetuation of influenza A viruses in Alaskan waterfowl reservoirs.
Ito T, Okazaki K, Kawaoka Y, Takada A, Webster RG, Kida H
Archeives of Virology, 140, 7, 1163, 1172, 1995年, ［査読有り］
英語, 研究論文（学術雑誌）

Brain lesions in young broiler chickens naturally infected with a mesogenic strain of Newcastle disease virus.
Bhaiyat MI, Ochiai K, Itakura C, Islam MA, Kida H
Avian pathology : journal of the W.V.P.A, 23, 693, 708, 4, 1994年12月, ［査読有り］

Mapping of transcriptional regulatory domains of pseudorabies virus immediate-early protein
S. Taharaguchi, H. Inoue, E. Ono, H. Kida, S. Yamada, Y. Shimizu
Archives of Virology, 137, 3-4, 289, 302, Springer-Verlag, 1994年09月, ［査読有り］
英語, 研究論文（学術雑誌）, The 180 kilodalton immediate-early protein (IE180) of pseudorabies virus functions as a strong transactivator of several different promoters and also as a repressor of its own transcription. To map the functional domains of IE180, we prepared various truncated mutants and analyzed their transcriptional regulatory activities using the chloramphenicol acetyl transferase (CAT) assay. Analysis of mutants truncated from the carboxy-terminal end of the 1460-amino acid polypeptide showed that a polypeptide possessing amino acids 1 to 1081 retained significant functions of transactivation and autoregulation potential. On the other hand, removing amino acids 1 to 131 resulted in a complete loss of transactivation potential, indicating that the domain responsible for transactivation is located in the amino-terminal end of IE180. Additional amino-terminal trunction up to amino acid 453 did not affect the autoregulation activity, indicating that the region between amino acids 454 and 1081 has autoregulation potential. © 1994 Springer-Verlag.

A NEUTRAL SUGAR IS RESPONSIBLE FOR SEROVAR SPECIFICITY OF THE ANTIGENIC DETERMINANT OF LEPTOSPIRA-INTERROGANS SEROVAR CANICOLA
H KASHIWASE, E ONO, R YANAGAWA, Y SHIMIZU, H KIDA
JAPANESE JOURNAL OF VETERINARY RESEARCH, 42, 2, 103, 108, HOKKAIDO UNIV, 1994年08月, ［査読有り］
英語, 研究論文（学術雑誌）, To provide information on the chemical structures of antigenic determinants of leptospira, glycolipids of Leptospira interrogans serovar canicola strain Hond Utrecht IV (Ut-IV) and its antigenic variant selected in the presence of a serovar-specific monoclonal antibody were compared physicochemically. Gas-liquid chromatography-mass spectrometry analysis revealed that the glycolipid of Ut-IV contained 6 neutral sugar species; rhamnose, mannose, galactose, glucose, and unknown sugars III and IV, in addition to unknown sugars I and II that had been previously reported. On the other hand, the glycolipid of the variant lacked unknown sugar III, suggesting that this sugar is responsible for the serovar-specific antigenic determinant.

Magnetic resonance imaging findings in Japanese encephalitis: White matter lesions
H. Shoji, H. Kida, H. Hino, S. Matsuura, K. Kojima, T. Abe, H. Utsunomiya, Y. Okada, Y. Nakamura, T. Okudera
Journal of Neuroimaging, 4, 4, 206, 211, Blackwell Publishing Inc., 1994年, ［査読有り］
英語, 研究論文（学術雑誌）, Ten patients with Japanese encephalitis diagnosed by serological criteria underwent magnetic resonance imaging (MRI) in axial and coronal sections. In 6, a second MRI study was done. The MRI findings were compared with the clinical outcome. Four patients died within several months of onset, 2 had sequelae such as hemiparesis and dementia, and the remaining 4 had no sequelae. In 9 of 10 patients, either diffuse or patchy white matter lesions were observed bilaterally, together with abnormalities in areas such as the thalamus, basal ganglia, and brainstem. For 3 patients who died or remained demented, the second MRI revealed extensive, diffuse white matter abnormalities. This study indicates that Japanese encephalitis can produce white matter involvement, although gray matter structures such as the thalamus, basal ganglia, and brainstem are more severely affected. The severity of these MRI lesions correlated with the clinical outcome.

A potential peptide vaccine against two different strains of influenza virus isolated at intervals of about 10 years.
Naruse H, Ogasawara K, Kaneda R, Hatakeyama S, Ito T, Kida H, Miyazaki T, Good RA, Onoe K
Proceeding of National Academy of Science, USA, 91, 20, 9588, 9592, 1994年, ［査読有り］
英語, 研究論文（学術雑誌）

Potential for transmission of avian influenza viruses to pigs.
Kida H, Ito T, Yasuda J, Shimizu Y, Itakura C, Shortridge KF, Kawaoka Y, Webster RG
Journal of General Virology, 75, 1, 2183, 2188, 1994年, ［査読有り］
英語, 研究論文（学術雑誌）

Acquisition of pathogenicity of a newcastle disease virus isolated from a Japanese quail by intracerebral passage in chickens.　　　　　　　　　　　　　　　
Islam MA, Ito T, Takakuwa H, Takada A, Itakura C, Kida H
Japanese Journal of Veterinary Research, 42, 3月4日, 147, 156, 1994年, ［査読有り］
英語, 研究論文（学術雑誌）

Antigenic drift of equine H3N8 influenza viruses.　　　　　　　　　　　　　　　
Kida H, Shimizu A, Ito T, Shimizu Y
Equine Infectious Disease VII, 307, 1994年, ［査読有り］
英語, 研究論文（学術雑誌）

Shortening of life expectancy in patients with membranous nephropathy — based on 20 years follow up study —
Makoto Ogi, Hiroshi Kida, Mitsuhiro Yoshimura, Yafumi Saito, Yoshinori Kibe, Gorou Sugioka, Kenzo Ikeda, Teruo Asamoto, Hitoshi Yokoyama, Naohisa Tomosugi
the japanese journal of nephrology, 35, 10, 1147, 1153, 1993年, ［査読有り］
英語, 研究論文（学術雑誌）, It is not certain whether the life expectancy of patients with membranous nephropathy is shorter than that of an age-matched healthy population. Forty-one patients (21 males, 20 females) aged between 16 and 70 years (average age: 33. 3 years) were followed for 20 years. The patients were divided into two groups: group I (n = 18), consisting of patients in whom nephrotic syndrome persisted for more than two years or until death, and group n (n = 23), consisting of patients except for group I. The non-survival criteria are death or renal death. Twelve patients (29. 3%) died during the study period. Eight patients belonged to group I and 4 to group n. The causes of death in group I patients were end-stage renal failure in 3 cases, ischemic heart disease in 1 case, subarachnoid hemorrhage in 1 case, malignancy in 2 cases, suicide in 1 case, and those in the group II patients were pneumonia, malignancy, cerebral softening, and diabetes mellitus, respectively. Eight patients who died in group I had a significantly longer difference between their actual life span (ALS) and life expectancy (LE) and a significantly smaller ratio of ALS to LE than the patients who died in group II (ALS—LE:—29. 9±4. 5 years in group I vs. -9. 0±6. 8 years in group II, p<
0. 05, ALS˟ 100/LE: 22. 5±8. 0% in group I vs. 80. 9±25. 2% in group II, p<
0. 05). In group I, the ratio of observed to expected death was 4. 76 (95% confidence interval, 2. 05 to 9. 37) and significantly higher than that of the control population. In group II, however, the ratio was 1. 09 (95% confidence interval, 0. 30 to 2. 80), and the difference from the control population was not statistically significant. These results suggest that longstanding nephrotic syndrome is associated with a shortened life expectancy in patients with membranous nephropathy. © 1993, Japanese Society of Nephrology. All rights reserved.

Adrenocortical carcinoma with a giant pericardial mass
Akio Nakata, Shinji Yagi, Koichi Oyama, Hiroshi Kida, Goro Sugioka
Internal Medicine, 32, 5, 438, 440, 1993年, ［査読有り］
英語, 研究論文（学術雑誌）, A patient with a history of adrenocortical carcinoma presented with massive pericardial effusion and a giant pericardial mass. Death resulted from cardiac tamponade. At autopsy a large necrotic mass was histologically similar to the original adrenocortical carcinoma. We present this case and review the literature on metastatic pericardial disease. (Internal Medicine: 32: 438-440, 1993). © 1993, The Japanese Society of Internal Medicine. All rights reserved.

A STRATEGY FOR MAKING SYNTHETIC PEPTIDE VACCINES
K OGASAWARA, H NARUSE, Y ITOH, T GOTOHDA, J ARIKAWA, H KIDA, RA GOOD, K ONOE
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 89, 19, 8995, 8999, NATL ACAD PRESS, 1992年10月, ［査読有り］
英語, 研究論文（学術雑誌）, We have determined the H-2 class II allele-specific amino acid motif of the agretope (the site of contact between the peptide antigen and the major histocompatibility complex) for a synthetic peptide composed of residues 43-58 of pigeon cytochrome c (p43-58). Residues 46 and 54 functioned as the agretope, and residues 50 and 52 functioned as the epitope (the site for contact between the peptide antigen and the T-cell antigen receptor). In general, agretopes and epitopes function independently. Thus, substitution of amino acids in the epitope does not significantly affect binding of the peptide antigen to a class II molecule. On the basis of these findings, synthetic peptide vaccines against influenza Aichi (H3N2) virus were prepared by introducing seven residues of the influenza virus hemagglutinin into the frame component residues 43-46 and 54-58 of p43-58 analogues including the agretopes for A(k) or A(b) previously determined on the p43-58 segment. These peptide vaccines induced both helper T-cell responses and production of antibodies that were specific for influenza Aichi hemagglutinin but not for the major histocompatibility complex binding frame in mice bearing A(k) or A(b). The antibodies produced neutralize the infectivity of influenza Aichi in vitro. The present findings should provide a basis for preparing potent peptide vaccines that function without producing side effects.

Effect of Sendai Virus Exposure on the Development of Frozen-Thawed Mouse Embryos
Carmencita Lavilla-Apelo, Antonio Arenas Rayos, Hiroshi Kida, Hiroshi Kanagawa
Journal of Reproduction and Development, 38, 3, 225, 228, 1992年, ［査読有り］
英語, 研究論文（学術雑誌）, Frozen-thawed mouse preimplantation embryos were exposed to Sendai virus at the blastocyst and morula stages to determine if these embryos were susceptible to virus infection. It was also examined whether the frozen-thawed embryos would further develop into expanded blastocysts after in vitro culture. Approximately 50% of the frozen-thawed embryos at the blastocyst stage were susceptible to Sendai virus infection. On the other hand, frozen-thawed embryos exposed to Sendai virus at the morula stage were resistant to the infection. It was noted that most of the frozen-thawed embryos exposed to Sendai virus developed into expanded blastocysts. Significant differences in the development to expanded blastocysts were observed in the Sendai virus-exposed frozen-thawed blastocysts (57.0%) compared with the frozen-thawed control (82.1%) and unfrozen control (93.7%) blastocysts. However, development to expanded blastocysts of Sendai virus exposed frozen-thawed morulae (73.0%) was similar to the frozen-thawed control (81.0%) but significantly different with the unfrozen control (93.2%) morulae. These findings suggest that some changes in the structure of the zona pellucida may occur during freezing procedures making the frozen-thawed embryos susceptible to virus infection. © 1992, THE SOCIETY FOR REPRODUCTION AND DEVELOPMENT. All rights reserved.

Effectiveness of steroid pulse therapy on progressive IgA nephropathy
Mitsuhiro Yoshimura, Hiroshi Kida, Yafumi Saito, Kihitiro Watanabe, Goro Sugioka
japanese journal of national medical services, 46, 8, 657, 663, 1992年, ［査読有り］
英語, 研究論文（学術雑誌）

A Patient with Asymptomatic Primary Biliary Cirrhosis Associated with Eosinophilic Infiltration and Peripheral Eosinophilia Improved By the Administration of Ursodeoxycholic Acid
Masakazu Yamazaki, Hideo Morimoto, Tokio Wakabayashi, Kunihiko Suzuki, Hiroshi Kida, Goto Sugioka
Kanzo, 33, 4, 348, 352, 1992年, ［査読有り］
英語, 研究論文（学術雑誌）, A 49-year-old male was admitted complaining of persistent cough, sputum and fever. The chest film examined on admission showed an infiltrative shadow in the right middle lung field. Blood tests revealed elevations of the ductal enzymes and a peripheral eosinophilia. However, the pulmonary infiltration with eosinophilia syndrome and drug-induced liver injury were negated by the absence of eosinophils in the lavaged fluids and in the lung specimens obtained by the transbronchial approach and by the negative lymphocyte-stimulation test against drugs prescribed for the respiratory symptoms. Further examinations revealed a positive anti-mitochondrial antibody and an increase in its M2 fraction measured at 46.7 U/ml. The diagnosis of primary biliary cirrhosis was suspected and was ascertained as Scheuer's stage I by the wedge liver biopsy, showing chronic non-suppurative destructive cholangitis (CNSDC) in the enlarged portal areas, which were markedly infiltrated with lymphocytes and plasma cells together with eosinophils. The eosinophilic infiltration was predominantly observed around the injured bile ducts. After the diagnosis was ascertained, ursodeoxycholic acid was given at a dose of 600 mg a day. Simultaneous improvements in the ductal enzymes and peripheral eosinophilia were observed one month later. The repeat biopsy carried out with an interval of 7 weeks after initiation of the treatment revealed only minor cellular infiltration without eosinophils and absent CNSDC. © 1992, The Japan Society of Hepatology. All rights reserved.

Steroid pulse therapy in rapidly progressive glomerulonephritis
Makoto Ogi, Mitsuhiro Yoshimura, Kenzo Ikeda, Yafumi Saito, Hiroshi Kida, Gorou Sugioka, Kihichirou Watanabe
The Japanese Journal of Nephrology, 34, 6, 669, 675, 1992年, ［査読有り］
英語, 研究論文（学術雑誌）, In an attempt to clarify the indication and efficacy of the methylprednisolone pulse therapy (1000mgX3times) for rapidly progressive glomerulonephritis (RPGN), 3 patients with the disease were carefully followed and the clinical course during and after the treatment were precisely analysed. According to the declination rate of reciprocals of serum creatinine(1/Cr), one patient were divided into the acute type (-1.00X 10”2dl/mg/day or less) and the others into the subacute type(more than -1.00 X 10∼2 dl/mg/day). In the patient of acute type, renal biopsy revealed cellular crescent formation in 93.8% of glomeruli observed. One course of the pulse therapy resulted in a decrease in Cr from 3.0mg/dl to 1.3mg/ dl and transformation of cellular crescents to fibrocellular or fibrous crescents. In the other two patients of subacute type, crescents were observed in 72.7% and 72.0% of glomeruli observed, and 87.5% and 38.9% of them were composed of cellular crescents respectively. Initial courses of the pulse therapy resulted in decreases of Cr from 3.5mg/dl to 2.4mg/dl and from 3.0mg/dl to 1.4mg/dl respectively. Additional courses, given because of insufficient reduction of Cr in the former, induced a further lowering to 1.3mg/dl, and because of re-elevation of Cr to 2.2mg/dl and remaining of cellular crescents in 20% in the latter, induced a decrease of Cr to 1.5mg/dl and disappearance of cellular crescents. © 1992, Japanese Society of Nephrology. All rights reserved.

THE EFFECTIVENESS OF TRYPSIN TREATMENT TO REMOVE SENDAI VIRUS ADHERING TO THE ZONA-PELLUCIDA OF MOUSE PREIMPLANTATION EMBRYOS
C LAVILLAAPELO, H KIDA, H KANAGAWA
JAPANESE JOURNAL OF VETERINARY RESEARCH, 39, 2-4, 133, 141, HOKKAIDO UNIV, 1991年12月, ［査読有り］
英語, 研究論文（学術雑誌）, The standard washing and trypsin treatment procedures to remove viruses adhering to the zona pellucida (ZP) were evaluated. Mouse embryos at the early blastocyst stage were exposed to Sendai virus, and then washed or treated with trypsin. Even after washing or trypsin treatment, Sendai virus was detected in the twelfth and final wash. The virus was still shown to adhere to the ZP by immunofluorescence assay. The embryos developed into expanded blastocysts following 24 hours of in vitro culture. Viral antigen was clearly demonstrated in the cells forming the expanded blastocysts, indicating that viral replication occurred in these cells. The present results suggest that the standard washing or trypsin treatment are not sufficient to remove Sendai virus adhering to the ZP of mouse embryos.

THE INFECTION OF MOUSE PREIMPLANTATION EMBRYOS TO SENDAI VIRUS (PARAINFLUENZA-I)
C LAVILLAAPELO, K OHTA, H KIDA, H KANAGAWA
THERIOGENOLOGY, 36, 1, 87, 94, BUTTERWORTH-HEINEMANN, 1991年07月, ［査読有り］
英語, 研究論文（学術雑誌）, To provide information on the susceptibility of mouse embryos to Sendai virus, it was investigated if viral replication occurs in the preimplantation embryo at different stages of development, with or without the zona pellucida (ZP). Mice were induced to superovulate, and embryos were collected on Days 2, 3 and 4 after mating. The ZP was removed by digestion with 0.5% pronase. Embryos were exposed to Sendai virus, washed, and allowed to develop in fresh culture medium. The presence of viral antigen in the embryonic cells was examined by the fluorescent antibody test (FAT). Specific immunofluorescence was demonstrated in the ZP-free morula and ZP-intact blastocyst. However, viral antigen was not detected in the ZP-intact two-cell, four-cell, eight-cell or morula stage embryos. Infected embryos developed normally to expanded blastocysts. These findings show that mouse embryonic cells are permissive hosts to Sendai virus replication and that the ZP played the role of a barrier against the virus.

Etiology of Rabbit Haemorrhagic Disease Spontaneously Occurring in Korea
J. H. Park, H. Kida, K. Ueda, K. Ochiai, M. Goryo, C. Itakura
Journal of Veterinary Medicine, Series B, 38, 1-10, 749, 754, 1991年, ［査読有り］
英語, 研究論文（学術雑誌）, Causative agent of rabbit haemorrhagic disease (RHD) was purified by CsCl density gradient centrifugation from the liver homogenate of rabbits infected with RHD virus which originated from Korea. The viral particles were 35–40 nm in diameter, and had hollow depressions on their surface. Protein A‐gold immunoelectron microscopy clearly showed that the convalescent antisera of diseased rabbits reacted specifically with the virus particles. SDS‐PAGE and Western blot analyses demonstrated that the structural protein of the virus was composed of a single major polypeptide of 63 kD. These findings indicate that the causative agent of RHD, tentatively named as picornavirus in Korea, belongs to calicivirus. © 1991 Blackwell Verlag GmbH

Influences of pregnancy on the natural course of chronic glomerulonephritis with impaired renal function
Shin-Ichi Takeda, Hiroshi Kida, Kazuya Takasawa, Kenzo Ikeda, MlTSUHIRO Yoshimura, HlTOSHI Yokoyama, Naohisa Tomosugi, Ken-ichi Kobayashi
The Japanese Journal of Nephrology, 33, 8, 803, 810, 1991年, ［査読有り］
英語, 研究論文（学術雑誌）, In an attempt to clarify the influence of pregnancy on the natural course of the chronic glomerulonephritis with impaired renal function (glomerular filtration rate : GFR&
iE
70ml/min), the courses of 14 pregnancies occurring in 10 patients (seven with IgA nephropathy, one with membranoproliferative glomerulonephritis, one with membranous nephropathy and one with hereditary nephropathy) were studied. In 8 patients GFR measured before pregnancies ranged from 46 to 70 ml/min and in the other two creatinine clearance estimated in the first trimester of pregnancies was 62 and 49 ml/min, respectively. The pregnancies resulted in 10 live births, one spontaneous abortion, one artificial abortion and 2 neonatal deaths. In 2 out of 10 live births fetal weight was less than 2500 g. In 3 of 11 pregnancies there was neither increase in urinary protein nor elevation of blood pressure during pregnancies, while seven (64:%) had increased proteinuria during the third trimester, and 4 of them were also complicated with hypertension. In 6 of 10 patients, there was no decrease in GFR during pregnancies. In three patients GFR was decreased from 70 to 36 ml/min, 70 to 58 ml/min and 62 to 48 ml/min, respectively. However, these reductions were considered to go with the natural course of respective patients because the reduction slopes were almost the same or rather mild in comparison with those estimated before or after pregnancies. The other patient also had a transient increase in serum creatinine level during two pregnancies, but the reciprocals of serum creatinine concentration before and after the pregnanciesdeclined linearly with time. These data suggest that pregnancy might have little influence on the natural course of the chronic glomerulonephritis even if complicated with renal functional impairment defined as GFR of 70 ml/min or less. © 1991, Japanese Society of Nephrology. All rights reserved.

Pathogenesis of diabetic nephropathy in non-insulin-dependent diabetes mellitus
Hiroshi Kida, Mituhiro Yoshimura, Kenzo Ikeda, Yafumi Saitou, Yutaka Noto
Journal of Diabetic Complications, 5, 2-3, 82, 83, 1991年, ［査読有り］
英語, 研究論文（学術雑誌）, To elucidate the characteristics of diabetic nodular lesions and the process of progression of diabetic glomerulosclerosis, kidney specimens obtained from 185 patients with non-insulin-dependent diabetes mellitus (NIDDM) were observed using light, electron, and immunofluorescence microscopes. The results suggest the following. First, there are two distinct subtypes of nodular lesions: One is formed by the progression and expansion of diffuse lesions
the other, showing a concentrically layered structure, is probably formed in the process of reconstruction of mesangiolysis. Second, there are three phases in the process of progression of diabetic glomerulosclerosis: In the first phase, arteriolosclerosis and diffuse lesions appear
in the second phase, mesangiolysis and nodular lesions develop in association with moderately advanced arteriolosclerosis
and in the third phase, exudative lesions and hyalinized glomeruli appear in association with advanced arteriolosclerosis together with advanced interstitial lesions. In the progression of the phases and in the development of mesangiolysis and layered nodular lesions, disturbed blood flow into glomeruli in consequence of diabetic arteriolosclerosis could be essential. © 1991.

A case of caroli's disease. the evaluation of MRI findings
Tokio Wakabayashi, Hideo Morimoto, Kunihiko Suzuki, Goroh Sugioka, Sakae Taira, Hiroshi Kida, Kishichiroh Watanabe
Kanzo, 32, 9, 884, 889, 1991年, ［査読有り］
英語, 研究論文（学術雑誌）, Although specific and pathogenetic findings of Caroli's disease on ultrasonography (US) and computed tomography (CT) have been already reported, magnetic resonance (MR) image of the disease has not been yet detailed, to our knowledge. Herein, we described characteristic MR findings found in a patient with the disease associated with congenital hepatic fibrosis, and compared with US and CT findings. Ultrasonography of the liver showed saccularly dilated intrahepatic bile ducts and intraluminal cord-like protrusions of the ductal walls, and contrast-enhanced CT demonstrated several tiny dots with strong contrast-enhancement within dilated bile ducts. By T2-weighted MR images, tiny dots without signal intensity were demonstrated. These were thought to be findings indicating portal radicles surrounded by dilated intrahepatic bile ducts, corresponding to the central dot sign by CT reported by Choi, et al (1990), and might be highly pathognomonic for the disease. We like to call the findings as the central flow void sign, and the sign could be very useful for the diagnosis of Caroli's disease. © 1991, The Japan Society of Hepatology. All rights reserved.

Molecular evidence for a role of domestic ducks in the introduction of avian H3 influenza viruses to pigs in southern China, where the A/Hong Kong/68 (H3N2) strain emerged.
Yasuda, J, Shortridge, K.F, Shimizu, Y, Kida, H
Journal of General Virology, 72, 8, 2007, 2010, 1991年01月, ［査読有り］
英語, 研究論文（学術雑誌）, The haemagglutinins (HAs) of five H3 influenza A viruses isolated from domestic ducks and one from a goose in southern China were analysed antigenically and genetically. The patterns of reactivity of two of the duck viruses and the goose virus with a panel of monoclonal antibodies to 10 different epitopes on the H3 HA were similar to those of influenza viruses isolated from wild ducks and pigs, as well as those of the earliest human H3 viruses. The other three isolates from domestic ducks were different from each other and from these viruses antigenically. Sequence analysis revealed that the HA genes of the two duck viruses and the goose virus were closely related to those of isolates from wild ducks and pigs; the identities between the deduced amino acid sequence of the HA of one of the isolates from domestic ducks and those of isolates from a wild duck and a pig were 98.7% and 99.5%, respectively. The antigenic and genetic similarity between these H3 HAs suggests that in southern China, the hypothetical influenza epicentre, domestic ducks may have played a role in the introduction of avian influenza viruses to pigs from feral ducks. The findings also support the hypothesis that the pig was a 'mixing vessel', producing a new human pandemic strain, A/Hong Kong/68 (H3N2), by genetic reassortment.

Acute toxoplasma myositis: an immunohistochemical and ultrastructural study
S. Matsubara, M. Takamori, H. Adachi, H. Kida
Acta Neuropathologica, 81, 2, 223, 227, Springer-Verlag, 1990年12月, ［査読有り］
英語, 研究論文（学術雑誌）, The occurrence is reported of acute myositis in a man with meningoencephalitis due to toxoplasmosis. The ultrastructure and immunohistochemistry of a muscle biopsy of the patient were investigated. Toxoplasma organisms were not found in the muscle biopsy. The perivascular inflammatory cells in the muscle were mainly CD4+ T cells and the inflammatory cells in and around the muscle fibres were chiefly macrophages. Expression of major histocompatibility complex class I and II antigens was observed in the infiltrating cells and endothelial cells of the blood vessels. A small proportion of the infiltrating cells expressed interferon-γ. A possible role of the immune mechanism in the evolution of myositis is discussed. © 1990 Springer-Verlag.

Primary glomerular fibrosis: a new nephropathy caused by diffuse intra-glomerular increase in atypical type III collagen fibers
K. Ikeda, H. Yokoyama, N. Tomosughi, H. Kida, A. Ooshima, K. Kobayashi
Clinical Nephrology, 33, 4, 155, 159, 1990年, ［査読有り］
英語, 研究論文（学術雑誌）, A 38-year-old Japanese man suffering from nephrotic syndrome and slowly progressive impairment of renal function underwent renal biopsy. Light microscopy revealed diffuse lobular accentuation of glomerular tufts, occupied by homogeneous material which was eosinophilic, lightly stained with pediodic acid silver methenamine and deeply with aniline blue. Further examinations using an electron microscope after treatment with phosphotungstic acid revealed that this material exhibited a diffuse increase in fibers having a periodicity of 65 ± 4 nm (mean ± SD) like type III collagen fibers and was reactive to the anti-type III collagen monoclonal antibody. In addition, the serum level of procollagen III peptides was markedly elevated to 221 ng/dl (normal range 2-13 ng/dl). However, the fibers were peculiarly curved and frayed as reported in the nail-patella syndrome, but were not observed in the lamina densa of the glomerular basement membrane (GBM) as seen in this syndrome. No family history or physical findings of bony and nail dystrophy, no laboratory data or pathological findings compatible with already described renal diseases including mesangiocapillary glomerulonephritis, light chain disease, immunotactoid nephropathy, amyloidosis, and non-amyloidotic fibrillary glomerulonephritis were observed. These data strongly suggest that the nephropathy is caused by a primary increase in atypical type III collagen fibers in glomeruli (primary glomerular fibrosis), which to our knowledge has not yet been described in the English literature.

A Case of Diffuse Panbronchiolitis(DPB) with Benign Monoclonal IgA Gammopathy and IgA Nephropathy with Monoclonal IgA Deposition
Takero Naito, Hitoshi Yokoyama, Yoshitaka Koshino, Naohisa Tomosugi, Nobu Hattori, Hiroshi Kida, Kunihiko Kobayashi
Japanese Journal of Medicine, 28, 4, 503, 505, 1989年, ［査読有り］
英語, 研究論文（学術雑誌）, A case of diffuse panbronchiolitis (DPB), associated with benign monoclonal IgA gammopathy and IgA nephropathy is described. The IgA deposition in the glomeruli of the patient was identified as consisting mainly of the monoclonal IgA having the same idiotype as that of serum monoclonal IgA. Recurrent infections of Hemophilus influenzae and Pseudomonas aeruginosa in the respiratory tract might have enhanced IgA-mediated immunity at the mucosal surface of the bronchiole, and ultimately induced monoclonal IgA gammopathy and IgA nephropathy. © 1989, The Japanese Society of Internal Medicine. All rights reserved.

Influence of pregnancy on IgA nephropathy
Shin-ichi Takeda, Hiroshi Kida, Kazuya Takasawa, Kenzo Ikeda, Masahiro Katagiri, Mitsuhiro Yoshimura, Hitoshi Yokoyama, Naohisa Tomosugi, Nobu Hattori
The Japanese Journal of Nephrology, 31, 1, 67, 75, 1989年, ［査読有り］
英語, 研究論文（学術雑誌）, In an attempt to clarify the influence of pregnancy on the natural course of IgA nephropathy, the courses of 79 pregnancies occurring in 47 patients with the disease were studied. These resulted in 3 artificial and 10 spontaneous abortions, and two pre-term and 64 full-term deliveries. Fifty four maternity passbooks were analyzed. In 22 pregnancies (40. 7%) proteinuria was increased during the third trimester, and in 13 (76. 5%) of 17 pregnancies receiving postpartum urinalysis, urinary protein was decreased to the level of the first trimester within one month after delivery. In two of the remaining four patients with a persistent increase in proteinuria, renal biopsy was carried out two months after delivery, revealing focal glomerular sclerotic lesions, in addition to mild mesangial proliferation compatible with IgA nephropathy. These findings indicated that increased urinary protein observed in the two pregnancies might be attributed to a complication of pre-eclamptic focal glomerular sclerosis rather than exacerbation of underlying IgA nephropathy. The glomerular filtration rate (GFR), examined in 27 patients both before and after pregnancy, was decreased in only two patients (7.4%), but these reductions appeared to go with the individual natural course. In 6 (15. 0%) of 40 pregnancies, proteinuria was increased within one month after delivery, and one of them was diagnosed both clinically and pathologically as the acute exacerbation of IgA nephropathy. These data suggest that patients with IgA nephropathy might show transient acute exacerbation just after delivery rather than during pregnancy, and that even if such exacerbations occurred, pregnancy might have little influence on the natural course of the disease. © 1989, Japanese Society of Nephrology. All rights reserved.

Origin of the hemagglutinin gene of A/Hong Kong/68 (H3N2) influenza virus.　　　　　　　　　　　　　　　
Kida H, Webster RG, Kawaoka Y, Shortridge KF, Naeve CV, Ito T, Itakura C, Mori F, Shimizu Y
Current Topics in Medical Virology, 365, 376, 1989年, ［査読有り］
英語, 研究論文（学術雑誌）

Clinicopathological characteristics of IgA nephropathy with acute onset: Comparative study with IgA nephropathy with insidious onset and purpura nephritis
Masahiro Katagiri, Hiroshi Kida, Takerou Naitou, Kazuya Takasawa, Shin-Ichi Takeda, Mitsuhiro Yoshimura, Hitoshi Yokoyama, Yoshitaka Koshino, Toshio Abe, Nobu Hattori
The Japanese Journal of Nephrology, 30, 1, 1, 7, 1988年02月01日, ［査読有り］
英語, 研究論文（学術雑誌）

Characteristics of Glomerular Lesions in Hepatitis B Virus Infection
Shin-ichi Takeda, Hiroshi Kida, Masahiro Katagiri, Hitoshi Yokoyarna, Toshio Abe, Nobu Hattori
American Journal of Kidney Diseases, 11, 1, 57, 62, 1988年, ［査読有り］
英語, 研究論文（学術雑誌）, In an attempt to clarify a participation of hepatitis B virus (HBV) in the development of hepatic glomerulopathy in adults, kidney specimens obtained from 151 patients with liver diseases were studied. Although mesangial proliferation was more severe in patients with chronic hepatitis or liver cirrhosis than in those with acute hepatitis, no significant difference was observed between 82 serum hepatitis B surface antigen (HBsAg)-positive (HBV-related group) and 69 negative patients (HBV-nonrelated group). However, double contours of the glomerular capillary walls were observed more often in the former group (18/82, P <
.01), especially in the HBeAg-positive subgroup (8/24, P <
.001), than in the latter (3/69). In addition, glomerular capillary spike formation or a bubblelike appearance was observed in seven patients of the former group. Of these, all five patients examined were HBeAgpositive in their serum. By electron microscopic studies, subendothelial dense deposits and mesangial interpositions were observed more frequently in the HBV related group, and subepithelial deposits were found only in the HBeAg-positive subgroup. The immunofluorescence study revealed IgA-dominant mesangial deposition in both HBV-related and nonrelated groups. As for the capillary wall deposits, however, IgG was dominant in 13 of the HBV-related group, but only one of the nonrelated group (P <
.01). Furthermore, one patient in the HBV-related group showed capillary wall-dominant HBeAg combined with IgG deposition. These results suggest that HBV-related glomerulopathy in adults may be characterized by capillary wall lesions, possibly attributed to deposition of immune complexes composed of HBV-related antigens, especially HBeAg and IgG class antibodies, and the glomerulopathy should be discriminated from so-called hepatic glomerulosclerosis. © 1988, National Kidney Foundation, Inc.. All rights reserved.

Identification of amino acids relevant to three antigenic determinants on the fusion protein of Newcastle disease virus that are involved in fusion inhibition and neutralization
T. Toyoda, B. Gotoh, T. Sakaguchi, H. Kida, Y. Nagai
Journal of Virology, 62, 11, 4427, 4430, 1988年, ［査読有り］
英語, 研究論文（学術雑誌）, Nucleotide sequence analysis of F protein antigenic variants of Newcastle disease virus mapped three distinct antigenic determinants to positions 343, 72, and 161 on the protein. The high fusion-inhibiting and neutralizing capacities of all of the monoclonal antibodies used for selection suggested close functional and structural relationships of the three positions with the fusion-inducing N-terminal region of the F1 subunit. The former two positions were located at the cysteine cluster domain near the C terminus of the F1 subunit and at the major hydrophilic domain in the F2 subunit, respectively, and both domains appeared to represent the major antigenic determinants of paramyxovirus F protein.

Mesangiolysis in diabetic glomeruli: Its role in the formation of nodular lesions
Y. Saito, H. Kida, S. I. Takeda, M. Yoshimura, H. Yokoyama, Y. Koshino, N. Hattori
Kidney International, 34, 3, 389, 396, 1988年, ［査読有り］
英語, 研究論文（学術雑誌）, In order to elucidate a participation of the mesangiolysis in the formation of diabetic nodular lesions, 355 kidney specimens obtained from 327 patients with primary diabetes mellitus were studied. Mesangiolyses begun by focal and segmental disintegration of the pivotal structure of the mesangium ('torn off phase'), resulting in cystic or aneurysmal dilatation of the involved tuft, were found in 56 specimens (16%). The dilated tufts were filled with lysed mesangial matrix, which showed a reticular or fibrillar arrangement ('structureless phase'), being followed by a concentrical re-arrangement ('reconstructive phase') and by the ultimate formation of diabetic nodules. The mesangiolysis of various phases was frequently found concomitant with severe diabetic arteriolosclerosis and, in the reconstructive phase, the lysed mesangial matrix near the recanalized capillary along the inner aspect of glomerular basement membrane was observed to be re-arranged in a layered structure. These results suggest the hypothesis that: 1) the mesangiolysis is the initial lesion occurring in glomeruli in the process of diabetic nodule formation, and disturbed blood flow into glomeruli, caused by diabetic arteriolosclerosis, may be implicated in the developoment of mesangiolysis
and 2) consequent compression of the lysed mesangial matrix by recanalized capillaries forms layered structures and ultimate completed diabetic nodules.

ORIGIN OF THE HEMAGGLUTININ GENE OF H3N2 INFLUENZA-VIRUSES FROM PIGS IN CHINA
H KIDA, KF SHORTRIDGE, RG WEBSTER
VIROLOGY, 162, 1, 160, 166, ACADEMIC PRESS INC JNL-COMP SUBSCRIPTIONS, 1988年01月, ［査読有り］
英語, 研究論文（学術雑誌）

ANTIGENIC AND GENETIC CONSERVATION OF H-3 INFLUENZA-VIRUS IN WILD DUCKS
H KIDA, Y KAWAOKA, CW NAEVE, RG WEBSTER
VIROLOGY, 159, 1, 109, 119, ACADEMIC PRESS INC JNL-COMP SUBSCRIPTIONS, 1987年07月, ［査読有り］
英語, 研究論文（学術雑誌）

Impaired immunoglobulin G production in minimal change nephrotic syndrome in adults
H. Yokoyama, H. Kida, T. Abe, Y. Koshino, M. Yoshimura, N. Hattori
Clinical and Experimental Immunology, 70, 1, 110, 115, 1987年, ［査読有り］
英語, 研究論文（学術雑誌）, Serum IgG, T and B cell subsets, cytoplasmic IgG positive cells (cBγ) and IgG in the medium (cIgG) of a 5 day culture of peripheral lymphocytes in both stimulated and non-stimulated (spontaneous) conditions with pokeweed mitogen (PWM) were studied in 30 adult patients with minimal change nephrotic syndrome (MCNS). In the nephrotic phase (11 patients), surface IgG positive B cells (sBγ) and spontaneous cBγ increased (P <
0.05), whereas PWM-stimulated cIgG did not increase, and serum IgG decreased significantly (P <
0.05). The cBγ/sBγ ratio calculated as an index of IgG synthesis in B cells increased spontaneously (P <
0.05), but did not increase under PWM-stimulation. The cIgG/cBγ ratio as an index of IgG secretion from each matured B cell, reduced in both spontaneous and stimulated conditions (P <
0.05, P <
0.01, respectively). In the phase of unstable remission maintained by steroid therapy (10 patients), these parameters tended to normalize and the OKT4/OKT8 ratio decreased (P <
0.05), while the ratio remained unchanged in the nephrotic phase. However, after discontinuation of steroid (nine patients), spontaneous cBγ and the spontaneous cBγ/sBγ ratio were again increased, and the cIgG/cBγ ratio decreased (P <
0.05) as observed in the nephrotic phase. These results suggest that B cells in patients with MCNS both in the nephrotic state and stable remission after discontinuation of steroid are activated spontaneously, but the secretory process of IgG from the matured cells is impaired, and that steroid improves these abnormalities.

Participation of renal tubular epithelial antigen and HLA-DR antigen in pathogenesis of membranous nephropathy
Hitoshi Yokoyama, Hiroshi Kida, Takero Naito, Kenzo Ikeda, Shin-Ichi Takeda, Mitsuhiro Yoshimura, Yoshitaka Koshino, Naohisa Tomosugi, Toshio Abe, Nobu Hattori
Japanese Journal of Nephrology, 29, 8, 995, 1002, 1987年, ［査読有り］
英語, 研究論文（学術雑誌）, In order to clarify the relationship between the tubulo-interstitial lesions and the subepithelial deposits along the glomerular capillary wall in membranous nephropathy (MN), participation of renal tubular epithelial antigen (RTE) and HLA-DR related antigen (Ial) in pathogenesis of the disease was studied. Twenty-nine biopsied and one autopsied kidney specimens obtained from 30 patients with MN (14 males and 16 females) were examined with the indirect immunofluorescence technique using monospecific rabbit anti-serum against human RTE and monoclonal anti-human Ial murine antibody. Fine granular deposits of RTE along the glomerular capillary wall was noted in five patients (17%), two with gouty kidney, one with malignant rheumatoid arthritis and two with idiopathic MN. These positive patients had moderate to severe tubulo-interstitial lesions. In 17 patiens (57%), Ial antigen was diffusely expressed in tubular epithelial ceils and the grade showed a positive correlation with the grade of tubulo-interstitial lesions (rs =0.579, p<
0.001). In addition, granular expression of Ial antigen in tubular epithelial cells was also observed in all RTE positive, but not in negative patients. Based on these results, it has been suggested that RTE may be involved in the pathogenesis in some patients with MN accompanied with tubulo-interstitial lesions and consequent aberrant Ial expression in tubular epithelial cells may play an important role in recognition of RTE as non-Self. © 1987, Japanese Society of Nephrology. All rights reserved.

Glomerular capillary wall immune deposits in IgA nephropathy and purpura nephritis
Mitsuhiro Yoshimura, Hiroshi Kida, Takerou Naitou, Satosi Goshima, Kazuya Takasawa, Shin-Ichi Takeda, Hitoshi Yokoyama, Yoshitaka Koshino, Toshio Abe, Nobu Hattori
Japanese Journal of Nephrology, 29, 8, 1077, 1085, 1987年, ［査読有り］
英語, 研究論文（学術雑誌）, In an attempt to clarify the participation of immune deposits in progression of glomerular lesions, relationship between electrondense deposits, especially subepithelial deposits observed by electron microscopy, and light microscopic and clinical findings in 131 patients with IgA nephropathy and 30 with purpura nephritis were studied. Capillary dense deposits together with mesangial deposits were observed in 16 (53%) patients with purpura nephritis in comparison with 46 (35%) with IgA nephropathy, especially subepithelial deposits being more frequently observed in purpura nephritis (15
50%) than in IgA rephropathy (23
18%) (p<
0.005). In addition, patients with subepithelial dense deposits showed heavier proteinuria with a mean of 1.9±0.2g/day in IgA nephropathy and of 2.0 ± 0.3 g/day in purpura nephritis, and higher incidence of cellular crescent (9
39 % and 12
80%, respectively). Furthermore, 5 out of 9 patients with IgA nephropathy, who were in an acute and active phase manifested by an abrupt appearance or increase in urinary protein and gross hematuria, showed subepithelial deposits. In purpura nephritis, 9 of 10 similar patients, all 8 with nephrotic syndrome and 3 with azotemia (Cr>
l,5mg/dl) disclosed subepithelial deposits. Although both daily urinary protein excretion and incidence of cellular crescents were higher in patients with purpura nephritis than those with IgA nephropathy, exacerbations scarcely occurred and prognosis was favourable in the former as compared to the latter, which showed a slowly progressive clinical course. These results suggest that subepithelial deposits develop in the acute and active phase in both diseases but cause different clinical courses as observed in the two diseases, favourable in purpura nephritis and progressive in IgA nephropathy, through difference of duration in disease activity, transient in the former and persistent in the latter. © 1987, Japanese Society of Nephrology. All rights reserved.

Re-pregnancy in patients with preeclamptic focal glomerular sclerosis
Kazuya Takasawa, Hiroshi Kida, Shin-Ichi Takeda, Satoshi Goshima, Kenzo Ikeda, Mitsuhiro Yoshimura, Hitoshi Yokoyama, Yoshitaka Koshino, Toshio Abe, Nobu Hattori
Japanese Journal of Nephrology, 29, 4, 401, 407, 1987年, ［査読有り］
英語, 研究論文（学術雑誌）, The possibility of re-pregnancy in patients having had focal glomerular sclerosis (FGS) and nephrotic syndrome caused by toxemia of pregnancy was studied. Renal biopsy performed in 20 patients with preeclampsia, in whom proteinuria of heavier than 1.0 g per day had persisted beyond one month after termination of the gestation, revealed findings compatible with FGS in 7 patients. Six of them achieved complete remission within 4 to 18 months and another patient did incomplete remission type I 12 months after delivery. With intervals of 9 to 29 months 3 patients in complete remission became re-pregnant, but proteinuria or hypertesion did not appear and the patients were able to have a baby with no impairment of renal function. These findings suggest that FGS developed during preeclampsia may follow a favorable clinical course and does not prevent re-pregnancy once complete remission has been achieved. © 1987, Japanese Society of Nephrology. All rights reserved.

Significance of IgA Deposits on the Glomerular Capillary Walls in IgA Nephropathy
Mitsuhiro Yoshimura, Hiroshi Kida, Toshio Abe, Shin-ichi Takeda, Masahiro Katagiri, Nobu Hattori
American Journal of Kidney Diseases, 9, 5, 404, 409, 1987年, ［査読有り］
英語, 研究論文（学術雑誌）, Based on immunofluorescence findings, 232 patients with IgA nephropathy were classified into two groups
one consisted of 88 patients (38%) with IgA deposits in the glomerular capillary walls together with the mesangial deposits (capillary type), and the other consisted of 144 patients (62%) with deposits confined to the mesangium (mesangial type). Electron microscopic findings revealed dense deposits on the capillary walls (subepithelial, 50%
intramembranous, 65%
and subendothelial, 24%) in 37 of 46 patients with capillary type and six of 47 with mesangial type (P <
.001). Crescent formation observed in ≥ 10% of glomeruli was more frequently found in patients with the capillary type (30/88, 34%) than those with the mesangial type (9/144,60/0) (P <
.01), especially higher in those with subepithelial deposits (15/26,57%). The capillary type patients showed heavier proteinuria (1.7 ± 0.2 g/d) than the mesangial type patients (0.6 ± 0.1 g/d) (P <
.05). Thirteen of the 14 patients in an acute exacerbation phase, manifested by an abrupt increase in urinary protein and development of macroscopic hematuria, showed capillary type IgA deposits. The ratio of patients with normal renal function in the fifth year after apparent onset was lower in the capillary type (74.0%) than in the mesangial type patients (96.9%) (P <
.05). These findings suggest that capillary IgA deposition is closely related to clinical and histologic activities of IgA nephropathy and is considered to be an important factor responsible for the progression of the disease, possibly through crescent formation. © 1987, National Kidney Foundation, Inc.. All rights reserved.

Effect of high-dose β-globulin therapy on membranous nephropathy
Naohisa Tomosugi, Hiroshi Kida, Hitoshi Yokoyama, Kenzo Ikeda, Yoshitaka Koshino, Eruo Asamoto, Toshio Abe, Nobu Hattori
Japanese Journal of Nephrology, 29, 1, 81, 86, 1987年, ［査読有り］
英語, 研究論文（学術雑誌）, Fifteen patients with idiopathic membranous nephropathy were treated with gammaglobulin (sulfonated or polyethylene glycol treated human gamma-globulin) at a daily dose of 10 g for 6 consecutive days. Two of them were treated twice with a one year interval. Follow up periods were 6 months in 3 patients and 12 months in 14 patients in all. After the initiation of the treatment, serum protein and albumin increased significantly, in an accompaniment with clinical improvement in 9 patients (53%) within 2 months and in 12 patients (71%) within 6 months. The OKT4/OKT8 ratio of peripheral blood, estimated serially in 14 patients using the flow cytometry, was initially 2.16±0.18, which was a significantly higher level than that of the controls (1.60±0.10, p<
0.01), but decreased to 1.61 ±0.17 in one month and maintained around same level throughout the period of 6 month observation, showing the ultimate ratio of 1.60±0.15 at the 6th month (p<
0.05), whereas it tended to rise one year later (1.77±0.27). These results indicate that the high-dose gamma-globulin administration causes reduction of OKT4/OKT8 ratio in membranous nephropathy and is useful for induction of early remission of the disease. © 1987, Japanese Society of Nephrology. All rights reserved.

SPIN-LABELING OF INFLUENZA-VIRUS HEMAGGLUTININ PERMITS ANALYSIS OF THE CONFORMATIONAL CHANGE AT LOW PH AND ITS INHIBITION BY ANTIBODY
S YODEN, H KIDA, M KUWABARA, R YANAGAWA, RG WEBSTER
VIRUS RESEARCH, 4, 3, 251, 261, ELSEVIER SCIENCE BV, 1986年05月, ［査読有り］
英語, 研究論文（学術雑誌）

Characteristics of glomerular lesions in hepatitis B virus infection
Shin-Ichi Takeda, Hiroshi Kida, Masahiro Katagiri, Katsumi Hirahara, Mitsuhiro Yoshimura, Hitoshi Yokoyama, Yoshitaka Koshino, Naohisa Tomosugi, Toshio Abe, Nobu Hattori
The Japanese Journal of Nephrology, 28, 4, 413, 420, 1986年, ［査読有り］
英語, 研究論文（学術雑誌）, In an attempt to clarify a participation of hepatitis B virus (HBV) in hepatic glomerular lesions in adults, 117 patients with liver disease were studied. They were divided into two groups
one including 69 patients positive for hepatitis B surface (HBs) antigen in serum (HBV related group), the other 48 negative for both HBs antigen and antibody (HBV non-related group). The former was further divided into 19 HBe antigen positive (HBeAg positive subgroup) and 12 HBe antibody positive patients (HBeAb positive subgroup). Although mesangial proliferation was more severe in patients with chronic liver disease (chronic hepatitis and liver cirrhosis) than with acute hepatitis, no significant differences were observed between the two groups and between the two subgroups. However, double contours of glomerular capillary walls were more often observed in HBV related group (17/69, 24.6%), especially HBeAg positive subgroup (8/19, 42.1%) than HBV non related group (3/48, 6.3%). Additionally, capillary spike-formation or bubble-like appearance was observed in 7 of 69 HBV related group (10.1%). Five of these were HBeAg positive, but the other two were not examined. Six of the 7 patients with spikes or bubbles were accompanied by double contours, but one without the double contour was diagnosed to have membranous nephropathy. In immunofluorescence study, IgA-predominant mesangial deposition was seen in both groups. However, in glomerular capillary walls, IgG equal or heavier deposition than IgA was observed in 13 of the 20 patients examined in HBV related group. These results suggest that hepatic glomerular lesions in adults may be consisted of the mesangial proliferation related to chronic liver disease and capillary lesions including double contours, spike-formation and bubble-like appearance, and in the development of the latter lesions HBV related antigens, especially HBeAg, and IgG class antibody may have an important role. © 1986, Japanese Society of Nephrology. All rights reserved.

Glomerular lesions in primary biliary cirrhosis
Yoshitaka Koshino, Hiroshi Kida, Kazuya Takasawa, Satoshi Goshima, Shin-Ichi Takeda, Mituhiro Yoshimura, Hitoshi Yokoyama, Naohisa Tomosugi, Toshio Abe, Nobu Hattori
The Japanese Journal of Nephrology, 28, 10, 1385, 1391, 1986年, ［査読有り］
英語, 研究論文（学術雑誌）, Glomerular lesions in patients with histologically proven primary biliary cirrhosis (PBC) were studied. Immunofluorescence examination revealed IgA deposition in 7 patients
both mesangium and glomerular capillary walls in 5, either of them in 2 patients. IgM was positive for the capillary wall in 6 patients in a fine granular pattern, grading from segmental to diffuse in distribution, concomitant with similar deposition of C3c in 5 patients. Five out of the 6 patients with mesangial IgA deposition were in advanced stages of PBC (Scheuer's stage III or IV) and the serum IgA level was elevated higher than 600 mg/dl in 4 of the 5 patients. However, there was no ralation between the capillary IgM deposition and the stage of PBC. The serum IgM level was elevated in all 8 patients, but there was no relation between IgM deposition and its serum level. Light microscopy revealed mild mesangial proliferation in 4 patients, but no capillay wall thickening, spike formation or bubble-like appearance was observed. Urinary protein was detected in only one patient averaging at 0.5g per day, who had mesangial IgA deposition but no IgM deposition. These data suggest that there are 2 types of glomerular immunoglobulin deposition in patients with PBC
one is the mesangial IgA deposition, probably due to chronic liver damage of PBC, and the other is the capillary IgM deposition, which may be characteristic for PBC. © 1986, Japanese Society of Nephrology. All rights reserved.

Participation of extracapillary lesions (ECL) in progression of IgA nephropathy
T. Abe, H. Kida, M. Yoshimura, H. Yokoyama, Y. Koshino, N. Tomosugi, N. Hattori
Clinical Nephrology, 25, 1, 37, 41, 1986年, ［査読有り］
英語, 研究論文（学術雑誌）, To clarify long-term prognosis and risk factors of IgA nephropathy, 205 patients with the disease were followed up for a period of 1 to 22 years with a mean of 7.9 years. According to the percentage of glomeruli involved with extracapillary lesions (ECL), which were defined by crescents and fibrous adhesion of glomerular tufts to Bowman's capsule, the patients were divided into four groups: group 1 - absence of ECL
group 2 - less than 25%
group 3 - 25-50%
group 4 - more than 50%. During the follow-up period, 26 patients progressed to chronic renal failure requiring hemodialysis, and one patient died of acute peritonitis. The actuarial kidney survival rate was 90.4% for 5 years and 86.0% for 10 years. Ten-year survival rates were 100% in group 1, 94.3% in group 2, 81.8% in group 3 and 25.5% in group 4. Re-biopsy specimens, observed in 31 patients after intervals of 1.4 to 13.4 years, revealed an increase of ECL in 20 patients concomitant with an apparent reduction of renal function in 13, whereas in the other 11 patients with no increase, renal function remained unchanged. These results strongly suggest that ECL plays an important role in the progression of IgA nephropathy, and renal function could be impaired by repeated formation and accumulation of these lesions.

ANTIGENIC MAPPING AND FUNCTIONAL-ANALYSIS OF THE F-PROTEIN OF NEWCASTLE-DISEASE VIRUS USING MONOCLONAL-ANTIBODIES
G ABENES, H KIDA, R YANAGAWA
ARCHIVES OF VIROLOGY, 90, 1-2, 97, 110, SPRINGER-VERLAG WIEN, 1986年, ［査読有り］
英語, 研究論文（学術雑誌）

Focal glomerular sclerosis in pre-eclampsia
H. Kida, S. Takeda, H. Yokoyama, N. Tomosugi, T. Abe, N. Hattori
Clinical Nephrology, 24, 5, 221, 227, 1985年, ［査読有り］
英語, 研究論文（学術雑誌）, Renal biopsy specimens of 16 patients with massive proteinuria, which developed during pregnancy and had lasted as late as one month after delivery, but disappeared within 15 months were studied. Four of them disclosed focal segmental hyalinosis, sclerosis, markedly swollen and foamy endocapillary cells, and ballooning of the involved tufts. Double contour of the capillary walls, predominance of epithelial cells as well as swelling of endocapillary cells known in pre-eclampsia were also observed. Electron and immunofluorescence microscopies revealed subendothelial deposition of IgM, C3 and rather weak fibrinogen-related antigens. These results indicate that focal glomerular sclerosis may develop during pre-eclamptic pregnancy, but may follow a non-progressive clinical course after delivery.

Organic glomerular lesions in toxemia of pregnancy
Shinichi Takeda, Hiroshi Kida, Mitsuhiro Yoshimura, Hitoshi Yokoyama, Yoshitaka Koshino, Teruo Asamoto, Naohisa Tomosugi, Toshio Abe, Nobu Hattori
The Japanese Journal of Nephrology, 27, 6, 803, 813, 1985年, ［査読有り］
英語, 研究論文（学術雑誌）, An attempt was made to clarify the characteristics of organic glomerular lesions precipitated by toxemia of pregnancy. Sixteen patients with moderate proteinuria (≧1.0 g/day) persisting for longer than one month following delivery but with no previous renal disease were studied, using light, electron (EM: 7/16) and immunofluorescence (IF: 10/16) microscopes. Renal biopsies performed 5 to 47 weeks (mean: 11 weeks) after delivery disclosed 3 distinct glomerular lesions, consisting of extracapillary, endocapillary and capillary lesions. A majority of the patients revealed 2 or 3 of these lesions simultaneously, but in each case one of these was highly predominant, so that the glomerular lesions could be classified into 4 histological types as follows: 1) Extracapillary type (6 cases): characterized by segmental crescentic proliferation of the epithelial cells. By IF study, IgG, C3 and fibrinogen (FRA) depositions were observed. EM findings revealed irregularity and extreme thinning of the glomerular basement membrane as well as subendothelial dense deposits. 2) Endocapillary type (4 cases): showed segmental hyalinosis, sclerosis, swollen endocapillary cells in foamy appearance and ballooning of the involved tufts. By IF study, IgM, C3 and FRA depositions were observed. 3) Capillary type (1 case): revealed a predominance of double contour of glomerular capillary walls. However, the remaining 5 cases showed minimal abnormalities. As for the clinical course, most patients had heavy proteinuria (≧3.5 g/day), which continued longer in patients with the extracapillary type lesions, while moderate proteinuria did not persist beyond 12 months with only one exception. Final GFR was improved by more than 80 ml/min. in all but one patient who had additional diffuse interstitial foam cells. From this study, it became evident that pre-eclampsia could precipitate organic glomerular lesions, consisting of 3 distinct lesions and forming 4 principal histological types, individuals of which followed a different clinical course, but overall prognosis of pre-eclamptic glomerulopathy might be favourable. © 1985, Japanese Society of Nephrology. All rights reserved.

Inhibition of transcriptase activity of influenza A virus in vitro by anti-haemagglutinin antibodies
Kazufumi Shimizu, Jun Mukaigawa, Mieko Oguro, Yasushi Ono, Katsuhisa Nakajima, Hiroshi Kida
Vaccine, 3, 3, 207, 210, 1985年, ［査読有り］
英語, 研究論文（学術雑誌）, An investigation was made of inhibition of transcriptase activity of influenza viruses in vitro by binding of antibody to the surface of the virion. Eight monoclonal antibodies which were directed against at least four non-overlapping antigenic regions of the haemagglutinin protein of A/Aichi/68 virus were tested for inhibitory effect. One of the antibodies directed against the B antigenic site, 22 1, inhibited transcriptase activity, while the other seven antibodies did not. Antibody from a hyperimmune rabbit serum to A/Udorn/72 (H3N2) virions inhibited the transcriptase activity of A/Udorn/72 and A/Aichi/68 (H3N2) viruses but not that of A/WSN/33 (H1N1). The antibody did not cause irreversible inactivation of the transcriptase since full activity was recovered by isolating ribonucleoprotein (RNP) cores from the inhibited virions using NP-40 treatment and subsequent centrifugation in a caesium sulphate density gradient. The antibody did not inhibit transcriptase activity of isolated RNP cores. The virion transcriptase activity was not inhibited by addition of the antiserum after the detergent treatment which is necessary for the activation of the transcriptase activity in vitro. These results suggest that the antibody blocks the activation process of the transcriptase by detergent treatment. © 1985.

Immunoglobulin-producing cells in minimal change nephrotic syndrome (MCNS)
Hitoshi Yokoyama, Hiroshi Kida, Yoshio tani, Yoshitaka Koshino, Shin-Ichi Takeda, Mitsuhiro Yoshimura, Naohisa Tomosugi, Toshio Abe, Nobu Hattori
The Japanese Journal of Nephrology, 27, 6, 773, 780, 1985年, ［査読有り］
英語, 研究論文（学術雑誌）, Dynamic changes of T and B lymphocyte subsets, and immunoglobulin (Ig) synthesis in cultured B lymphocytes were studied in accordance with the clinical activity in 29 adult patients with minimal change nephrotic syndrome (MCNS). The subsets (T3, T4, T8, sB
', sBa, sB/i, and sBs) were identified by detection of cell surface markers such as 0KT3, OKT 4, OKT8, and j, a, fi and s chain of Ig respectively. Cytoplasmic Ig positive cells (cB) were studied after culture of peripheral lymphocytes for 5 days. In the nephrotic phase (n=10), T lymphocyte subsets remained unchanged, but sB (sB
', sBa, sB/i, sBs) and spontaneous cB (cB]', cB, cBs) were increased. However, cB induced by pokeweed mitogen (PWM: 10/ig/ml) did not differ significantly from the control levels, and the PWM stimulation index (St. I. =cB with PWM/cB without PWM) in the IgG produ-3 cing system was decreased (p<
0.005). In the remitting phase by steroid therapy (n = 10), T3, T4 and T4/T8 (helper/suppressor) ratio revealed a significant reduction in an accompaniment with a tendency of normalization of B lymphocyte subsets. However, spontaneous and PWM induced cB were increased no more than the control value, resulting in a significant reduction of cB/sB ratio. In the remitted phase requiring no steroid therapy (n=9), all of the parameters were normalized except for the PWM St. I. in the IgG producing system, which was decreased (p<
0. 025). Autologous plasma of nephrotic phase significantly reduced cB with or without PWM, but remitted plasma did not. These results suggest that in the nephrotic phase of MCNS B lymphocyte subsets are already activated in vivo, but their proliferation or maturation from sB to cB is suppressed. © 1985, Japanese Society of Nephrology. All rights reserved.

Immunodynamics of focal glomerular sclerosis
Hitoshi Yokoyama, Hiroshi Kida, Tomosugi Aohisa, Masahiro Katagiri, Katsumi Hirahara, Shin-ichi Takeda, Mitsuhiro Yoshimura, Yoshitaka Koshino, Toshio Abe, Nobu Hattori
The Japanese Journal of Nephrology, 27, 8, 1095, 1101, 1985年, ［査読有り］
英語, 研究論文（学術雑誌）, Dynamic changes of T lymphocyte subsets and serum immunoglobulin (Ig) E levels were studied in focal glomerular sclerosis (FGS I 8 cases) and minimal change nephrotic syndrome (MCNS I 11 cases). Twenty-two healthy volunteers were served as a control group. T lymphocyte subsets were identified by the indirect immunofluorescence technique using monoclonal antibodies
OKT 3 (T 3
peripheral T), OKT 4 (T 4
helper/inducer) and OKT 8 (T 8
suppressor/cytotoxic). Serum IgE was measured by the radioimmunosorbent assay. All patients were treated with prednisolone in a daily dose of 40 mg or 60 mg for 4 to 8 weeks, follow by maintenance dose of 10 to 20 mg a day for at least 2 years. Patients with FGS were clinically separated into 2 groups, according to clinical status in the fourth month after initiation of the steroid therapy. In group I, 4 patients who were responded to the therapy, resulting in disappearance of proteinuria were included, and 4 patients in group II failed to response, remaining in nephrotic state or incomplete remission (urinary protein excretion, T. 0 g/day) through the initial 4 months. In the nephrotic phase of FGS and MCNS, serum IgE levels were markedly elevated but there was no significant change of T lymphocyte subsets. After administration of steroid, T 3, T 4, and T4/T8 ratio were decreased, and serum IgE returned to the normal range in group I. However, in group II, T3, T4 and T4/T8 ratio were increased, and serum IgE was decreased but still remained in a higher level, MCNS showed decreases in T 3, T 4, T4/T8 ratio and serum IgE and increase in T 8 after steroid therapy as observed in group I of FGS. This study clarified that FGS could be separated into two groups based on the clinical response to steroid therapy, and each groups might have individual response pattern of T lymphocyte subsets, suggesting an important role of the response pattern to determine a prognosis in patients with FGS. © 1985, Japanese Society of Nephrology. All rights reserved.

Survival of Corynebacterium renale, Corynebacterium pilosum and Corynebacterium cystitidis in soil
Aoi Hayashi, Ryo Yanagawa, Hiroshi Kida
Veterinary Microbiology, 10, 4, 381, 386, 1985年, ［査読有り］
英語, 研究論文（学術雑誌）, Survival of the causative agents of bovine pyelonephritis, Corynebacterium renale, C. pilosum and C. cystitidis, was examined at 30°C in autoclaved soil. In the soil from a paddock, C. renale and C. cystitidis survived for 56 and 63 days, respectively, and C. pilosum for a longer period of at least 210 days. In soil from a pasture, sand from an athletic field and sea sand, the survival of these bacteria was of shorter duration. © 1985.

Adhesion of Corynebacterium renale and Corynebacterium pilosum to the epithelial cells of various parts of the bovine urinary tract from the renal pelvis to vulva
Aoi Hayashi, Ryo Yanagawa, Hiroshi Kida
Veterinary Microbiology, 10, 3, 287, 292, 1985年, ［査読有り］
英語, 研究論文（学術雑誌）, The various parts of the bovine urinary tract, the renal pelvis, the ureter, the urinary bladder, the urethra, the vaginal vestibule and the vulva, were examined for the capacity of the epithelial cells to bind Corynebacterium renale and C. pilosum. C. renale adhered best to the epithelial cells of the vulva, and then to those of the ureter and renal pelvis. C. pilosum also adhered best to the epithelial cells of the vulva, followed by those of the vaginal vestibule. The results indicate that the most important target tissue for these bacteria may be the vulva, and the results correlate with the fact that C. renale frequently causes pyelonephritis and ureteritis, while C. pilosum causes the same diseases less frequently and behaves like normal flora of the vaginal vestibule. © 1985.

IS BIVALENT BINDING OF MONOCLONAL-ANTIBODIES TO DIFFERENT ANTIGENIC AREAS ON THE HEMAGGLUTININ OF INFLUENZA-VIRUS REQUIRED FOR NEUTRALIZATION OF VIRAL INFECTIVITY
S YODEN, H KIDA, R YANAGAWA
ARCHIVES OF VIROLOGY, 85, 3-4, 209, 216, SPRINGER-VERLAG WIEN, 1985年, ［査読有り］
英語, 研究論文（学術雑誌）

INTERFERENCE WITH A CONFORMATIONAL CHANGE IN THE HEMAGGLUTININ MOLECULE OF INFLUENZA-VIRUS BY ANTIBODIES AS A POSSIBLE NEUTRALIZATION MECHANISM
H KIDA, S YODEN, M KUWABARA, R YANAGAWA
VACCINE, 3, 3, 219, 222, BUTTERWORTH-HEINEMANN LTD, 1985年, ［査読有り］
英語, 研究論文（学術雑誌）

DETECTION OF ANTIBODIES TO LEPTOSPIRAL GENUS-SPECIFIC ANTIGEN IN HUMAN AND ANIMAL SERA BY INDIRECT HEMAGGLUTINATION TEST WITH A PARTIALLY PURIFIED GENUS-SPECIFIC PROTEIN ANTIGEN
N SAKAMOTO, R YANAGAWA, E ONO, H KIDA, M MORI, Y ARIMITSU, K AKAMA, J YASUDA, K TOO
ZENTRALBLATT FUR BAKTERIOLOGIE MIKROBIOLOGIE UND HYGIENE SERIES A-MEDICAL MICROBIOLOGY INFECTIOUS DISEASES VIROLOGY PARASITOLOGY, 259, 4, 548, 556, GUSTAV FISCHER VERLAG, 1985年, ［査読有り］
英語, 研究論文（学術雑誌）

PRODUCTION OF MONOCLONAL-ANTIBODIES AGAINST LEPTOSPIRAL GENUS-SPECIFIC PROTEIN ANTIGEN AND LOCALIZATION OF THE ANTIGEN BY IMMUNOELECTRON MICROSCOPY
N SAKAMOTO, E ONO, H KIDA, R YANAGAWA
ZENTRALBLATT FUR BAKTERIOLOGIE MIKROBIOLOGIE UND HYGIENE SERIES A-MEDICAL MICROBIOLOGY INFECTIOUS DISEASES VIROLOGY PARASITOLOGY, 259, 4, 557, 563, GUSTAV FISCHER VERLAG, 1985年, ［査読有り］
英語, 研究論文（学術雑誌）

Antigenic analysis of H4 influenza virus isolates using monoclonal antibodies to defined antigenic sites on the hemagglutinin of A/budgerigar/Hokkaido/1/77 strain.　　　　　　　　　　　　　　　
Ito T, Kida H, Yanagawa R
Archeives of Virology, 84, 251, 259, 1985年, ［査読有り］
英語, 研究論文（学術雑誌）

The Significance of Predominant IgM Deposition in Glomerular Diseases
Yoshitaka Koshino, Hiroshi Kida, Hitoshi Yokoyama, Mitsuhiro Yoshimura, Teruo Asamoto, Yafumi Saito, Naohisa Tomosugi, Toshio Abe, Nobu Hattori
The Japanese Journal of Nephrology, 26, 10, 1293, 1300, 1984年, ［査読有り］
英語, 研究論文（学術雑誌）, Ninety-four of 994 renal biopsy specimens observed under both light and immunofluorescence microscopes revealed predominant or sole immunoglobulin M (IgM) deposition in glomeruli. These were minimal change nephrotic syndrome (MCNS) in 22, focal glomerular sclerosis (FGS) in 11, mesangial proliferative glomerulonephritis in 21, membrano-proliferative glomerulonephritis in 8 and other diseases, but none was not able to be diagnosed using already established definitions of renal diseases. As for MCNS and FGS, IgM deposition was observed in 27 out of 56 (47%) patients with MCNS mainly in mesangial areas and in 11 out of 15 (73%) patients with FGS mainly in mesangial areas in four and mainly in the capillary walls in the other seven patients. Electron microscopic examination performed on eight patients with MCNS and IgM deposition disclosed dense mesangial deposits in three patients. Simultaneous deposition of C3c with IgM was found in 18 out of 27 (67%) patients with MCNS and in 7 out of 11 (64%) patients with FGS. However, no statistical difference were present between those with glomerular IgM deposition and those without in MCNS and FGS, concerning resposiveness to steroid treatment, incidence of relapse and outcome of renal function. Six patients with MCNS and IgM deposition had repeat biopsy, which revealed no progression to FGS. From this study it become evident that predominant or sole IgM deposition can be seen in various glomerular diseases and IgM deposition in MCNS or FGS did not predict a poor response or outcome. Thus, we conclude that IgM deposition in glomeruli has no relation to specific glomerular lesions or clinical courses. © 1984, Japanese Society of Nephrology. All rights reserved.

Intermediate type of primary glomerulonephritis between mesangial proliferative and membranoproliferative glomerulonephritis
Yoshio Tani, Hiroshi Kida, Toshio Abe, Naohisa Tomosugi, Yafumi Saito, Teruo Asamoto, Yoshitaka Koshino, Hitoshi Yokoyama, Mitsuhiro Yoshimura, Nobu Hattori
The Japanese Journal of Nephrology, 26, 10, 1337, 1344, 1984年, ［査読有り］
英語, 研究論文（学術雑誌）, Ten patients with primary mesangial proliferative glomerulonephritis (Mes PGN) with mild, irregular but distinct thickening of glomerular capillary walls observed by the light microscopy were studied. Silver staining revealed mesangial interposition and spike formation as well as apparent but not severe mesangial proliferation in all patients, but none of them showed lobular formation of glomerular tufts. Mesangial interposition were estimated :as circumferential and diffuse similar to membranoproliferative glomerulonephritis (MPGN) type in in two patients, but in the other eight the interposition was only partial, so that the diagnosis of MPGN was considered to be improper for them. Electron microscopic xammation, performed in seven patients, revealed mild to moderate deposition of subepithelial, subendothelial and mesangial dense deposits in almost equal severity. However, the immunofluorescence study for immunoglobulins (Ig) and C3 were rather predominant in the capillary wall than in the mesangial area and predominance was found in IgG in seven and in C3 in three patients. Their initial symptoms were acute nephritic syndrome (1), edema (1) and chance proteinuria (8). Their initial urinary protein was rather prominent compared to individual mesangial proliferation. Nephrotic syndrome appeareed in three patients in their clinical course but did not last longer than 6 months. None of them showed persistent hypocomplementemia. During the follow-up period, which ranged from 14 to 121, with a mean of 56 months, only one had fallen into chronic renal failure requiring regular hemodialysis after a 22 year clinical course and seven patients have kept. © 1984, Japanese Society of Nephrology. All rights reserved.

Renal pathology in fulminant hepatitis : Tubulointerstitial lesions accompanied with multinucleated giant cells
Hitoshi Yokoyama, Hiroshi Kida, Mitsuhiro Yoshimura, Yoshitaka Koshino, Teruo Asamoto, Yafumi Saito, Naohisa Tomosugi, Toshio Abe, Nobu Hattori
The Japanese Journal of Nephrology, 26, 11, 1479, 1485, 1984年, ［査読有り］
英語, 研究論文（学術雑誌）, A patient with fulminant hepatitis and multinucleated giant cells in renal tubulointer-stitium was reported, and further study on renal pathology in fulminant hepatitis was attempted on 23 autopsy cases. A 48-year-old man, diagnosed as fulminant hepatitis based on severe jaundice and disturbed consciousness, was treated with dexamethasone, amino acid solution, insulin-glucagon therapy and plasma exchange, but both serum bilirubin and creatinine were increased to 23.7 mg/dl and 3.4 mg/dl, respectively, at the sixth hospital week. Four weeks later he died of pneumonia when serum bilirubin was read at 40. 9 mg/ dl. Autopsy revealed submassive hepatic necrosis and acute bronchopneumonia. As for renal lesions, glomeruli and vessels appeared almost normal, but in tubules tubular epithelial cell degeneration, bile casts formation and multinucleated giant cells, which contained bile stained granules and bile casts, were found. Some of the giant cells seen in the interstitium were surrounded with infiltrated small round cells and formed granulomatous lesions. By the way, in 23 patients with fulminant hepatitis. Proteinuria developed in 7, microscopic hematuria in 5 and azotemia in 8 patients. Histological examinations disclosed mild glomerular lesions in 11, degenerative tubular lesions in 11, bile casts in 7 and intratubular multinucleated giant cells phagocytosing bile casts in 4 patients. The seven patients having bile casts or giant cells showed higher serum direct bilirubin (26. 1 + 3. 2 mg/dl, mean � S. E. M. ) and creatinine (3. 06 + 0. 60 mg/dl) levels, compared with the other 16 patients (10. 4 + 1. 6 mg/ dl, p<
0.005
0. 88 + 0.18 mg/dl, p<
0.001). These results suggest that fulminant hepatitis frequently accompanied with renal lesions including giant cells probably formed from renal tubular epithelial cells in a process of treatment of bile casts. To the best of our knowledge, this is the first report of fulminant hepatitis accompanied with tubulointerstitial multinucleated giant cells. © 1984, Japanese Society of Nephrology. All rights reserved.

Mesangiolysis in diabetic glomerulosclerosis
Yafumi Saito, Hiroshi Kida, Mitsuhiro Yoshimura, Hitoshi Yokoyama, Yoshitaka Koshino, Teruo Asamoto, Naohisa Tomosugi, Toshio Abe, Nobu Hattori
The Japanese Journal of Nephrology, 26, 4, 367, 375, 1984年, ［査読有り］
英語, 研究論文（学術雑誌）, Three hundred kidney biopsy specimens and 10 autopsy specimens were reviewed with an attempt to describe diabetic ��mesangiolysis (ML)� and to clarify its role in a development of diabetic glomerulosclerosis, especially of intraglomerular microaneurysms and of nodular lesions. ML observed under a light microscope was segmental and centrilobular meshwork-making, lamellated, acellular or less-cellular amorphous and disorganized lytic lesions of mesangium and was usually accompanied with microaneurysm (88%), formed from dilated capillaries surrounding the mesangiolytic area and with foam cells (51%) located in the periphery of the tufts. The lesion was observed in 41 of 59 specimens (69%) with nodular lesions, compared with only 2 out of 251 specimens (0.8%) without nodular lesions, but in 21 of 310 specimens all of ML, nodular lesions, microaneurysms and foam cells were observed simultaneously. In some cases, however, the �meshwork-making� or lamellated materials in the mesangiolytic area were hardly distinguished from a small diabetic nodule. The incidence of all these lesions present in diabetic glomeruli were increased in accordance with progression of diffuse lesions and diabetic arteriolo-clerosis. From these results, it is suggested that ML, microaneurysms and nodular lesioris are coincidental lesions and that ML may be an initial insult in forming microaneurysms and nodular lesions in diabetic glomeruli. © 1984, Japanese Society of Nephrology. All rights reserved.

Human influenza virus infection in mink: serological evidence of infection in summer and autumn.
Okazaki K, Yanagawa R, Kida H, Noda H
Veterinary microbiology, 8, 251, 257, 3, 1983年06月, ［査読有り］

INHIBITION OF VIRUS-INDUCED HEMOLYSIS WITH MONOCLONAL-ANTIBODIES TO DIFFERENT ANTIGENIC AREAS ON THE HEMAGGLUTININ MOLECULE OF A SEAL MASSACHUSETTS 1/80 (H7N7) INFLUENZA-VIRUS
H KIDA, RG WEBSTER, R YANAGAWA
ARCHIVES OF VIROLOGY, 76, 2, 91, 99, SPRINGER-VERLAG WIEN, 1983年, ［査読有り］
英語, 研究論文（学術雑誌）

Contact infection of mink with 5 subtypes of avian influenza virus. Brief report.
Okazaki K, Yanagawa R, Kida H
Archives of virology, 77, 265, 269, 2-4, 1983年, ［査読有り］

Presence of Vi antigen in a virulent strain of Leptospira interrogans serovar pomona and relation of Vi antigens of leptospiras to resistance to leptospiricidal activity mediated by antiserum plus complement
K. Ueno, R. Yanagawa, H. Kida
Zentralblatt fur Bakteriologie Mikrobiologie und Hygiene - Abt. 1 Orig. A, 252, 4, 557, 565, 1982年, ［査読有り］
英語, 研究論文（学術雑誌）, Vi antigen was found in a virulent strain of Leptospira interrogans serovar pomona. The presence of Vi antigen was shown by the agglutinin-absorption and precipitin-absorption tests. Each strain of pomona and copenhageni which possessed Vi antigen and was resistant to the leptospiricidal activity mediated by the antiserum plus complement lost Vi antigen and became susceptible to the leptospiricidal activity when cultured in the presence of 8-azaguanine. The strains thus treated with 8-azaguanine recovered the resistance and Vi antigen when cultured in medium without 8-azaguanine.

The Clinical Significance of Crescents in Idiopathic Mesangial Proliferative Glomerulonephritis
Toshio Abe, Hiroshi Kida, Hitoshi Yokoyama, Yoshitaka Eoshino, Teruo Asamoto, Yafumi Saito, Yoshio Tani, Naohisa Tomosugi, Nobu Hattori
The Japanese Journal of Nephrology, 24, 9, 987, 995, 1982年, ［査読有り］
英語, 研究論文（学術雑誌）, Clinical significance of crescent formation especilly in less than 50% of glomeruli have been unsatisfactorily studied. Thus, we attempted to clarify this point, and investigated the clinical features and the prognoses of 53 patients with mesangial proliferative glomerulonephritis with cellular or fibrocellular crescents in more than 10% of glomeruli. Fourteen patients having more than 50% of glomeruli involved by crescent formation (group I) were analyzed separately from 39 patients with less than 50% involvement (group II). In group I, 35% (12/14) of patients presented acute onset and reduced renal function (GFR>
80m//min). Renal function was subsequently declined rapidly and 12 oLl4 patients progressed to terminal renal failure. In group n, 77% (30/39) of patients presented incidious onset and 54% (21/39) of patients presented normal renal function. Even in this group, however, renal function was declined slowly but steadily, and 10 of 39 patients progressed to renal failure. The survival rates of group I were 31% at a 5 years and 15% at 10 years. Those of group II were 75% and 63%, respectively. The extent of inter-stitium involved by cellular infiltration revealed good relation to the prognosis and the percentage of glomeruli involved by crescent. From this study, it is suggested that the clinical feature and prognosis are influenced by the percentage of glomeruli involved by crescent. And it is proven that glomerulonephritis with crescent formation even in less than 50% of glomeruli was progressive, as well as it in more than 50%. © 1982, Japanese Society of Nephrology. All rights reserved.

BIOLOGICAL-ACTIVITY OF MONOCLONAL-ANTIBODIES TO OPERATIONALLY DEFINED ANTIGENIC REGIONS ON THE HEMAGGLUTININ MOLECULE OF A/SEAL MASSACHUSETTS/1/80 (H7N7) INFLUENZA-VIRUS
H KIDA, LE BROWN, RG WEBSTER
VIROLOGY, 122, 1, 38, 47, ACADEMIC PRESS INC JNL-COMP SUBSCRIPTIONS, 1982年, ［査読有り］
英語, 研究論文（学術雑誌）

PRESENCE OF VI ANTIGEN IN A VIRULENT-STRAIN OF LEPTOSPIRA INTERROGANS SEROVAR POMONA AND RELATION OF VI ANTIGENS OF LEPTOSPIRAS TO RESISTANCE TO LEPTOSPIRICIDAL ACTIVITY MEDIATED BY ANTISERUM PLUS COMPLEMENT
K UENO, R YANAGAWA, H KIDA
ZENTRALBLATT FUR BAKTERIOLOGIE MIKROBIOLOGIE UND HYGIENE SERIES A-MEDICAL MICROBIOLOGY INFECTIOUS DISEASES VIROLOGY PARASITOLOGY, 252, 4, 557, 565, GUSTAV FISCHER VERLAG, 1982年, ［査読有り］
英語, 研究論文（学術雑誌）

Membranoproliferative Glomerulonephritis and Allied Glomerulonephritis
Kazuhiro Dohi, Yasushi Nakamoto, Hiroshi Kida, Toshio Abe, Masahiko Fujioka, Nobu Hattori
The Japanese Journal of Nephrology, 23, 2, 195, 207, 1981年, ［査読有り］
英語, 研究論文（学術雑誌）, This report deals with the development and prognosis of, and some effects of treatment on membranoproliferative glomerulonephritis (MPGN) and allied clomerular chances. The subjects consisted of 24 patients (15 males and 9 females) with ages of 11 to 79 years (an average of 27.8). The glomerular pathology was classified into lobular form (2 cases), classical form (9) and atypical form (13). Lobular form is consistent with “lobular glomerulonephritis” described by Allen, and classical form with “pure form” termed by Habib, while atypical form is incomplete to be called classical form, since glomeruli with classical form were observed in a focal distribution (50-75% of glomeruli obtained), or circumferential mesangial interposition was present in a local fashion (50-75% of the glomerular tuft). The present series did not include “dense deposit disease ”, and belonged to type I variety of MPGN in all but one case which was type III. Three modalities of treatment were at random performed on 17 cases, i.e., steroids and/or cyclophosphamide, anticoagulants and/or fibrinolytics, and both of them (combined therapy) on 5, 4 and 8 cases, respectively. All 8 patients given combined therapy showed stable or improved renal functions. On the other hand, deterioration of renal function was observed in 2 out of 5 cases with steroids and/or cyclophosphamide, 3 out of 4 with anticoagulants and/or fibrinolytics, and 3 out of 7 untreated patients. In relation to the glomerular pathology, atypical form was the most responsive to any mode of treatment. Six out of 14 cases with repeat or serial renal biopsies developed the histological transitions
1) from mild to moderate mesangial proliferative GN to atypical form (2 cases), 2) from acute proliferative and exudative GN of nonstreptococcal origin, to atypical form. 3) from atypical form to classical one, 4) from classical form to lobular form, 5) from atypical form to mild to moderate mesangial proliferative GN after treatment. Overall outcome was as follows. Two cases of lobular form progressed to require hemodialysis or died within 15±10 months after the clinical onset. Five patients with classical form are alive and the remaining 4 died of uremia during 88.3±51.6 months. In atypical form, only one case underwent hemodialysis and other 12 are alive with good functions in the period of 101.9±61.3 months. Thus, the following conclusions can be drawn from the present study. 1) The more severe hyperplasia of mesangial matrix, the poorer outcome could be expected, with lobular form carrying the most ominous prognosis. 2) At the moment, combined therapy over years is, seemingly, worthy of trial, especially at a stage of atypical form to prevent further histological development. 3) The transition of classical to lobular form offers an additional support for the intimate relationship between MPGN and lobular GN. © 1981, Japanese Society of Nephrology. All rights reserved.

SURVEY OF INFLUENZA-VIRUSES IN FERAL BIRDS IN 1979 AND ISOLATION OF A STRAIN POSSESSING HAV6NAV5 FROM CLOACA OF AN EASTERN DUNLIN
E HONDA, H KIDA, R YANAGAWA, Y MATSUURA, K YAGYU, M TSUJI, K UENO, N YAMAUCHI, S MISHIMA, H OGI, K SHIMAZAKI
JAPANESE JOURNAL OF VETERINARY RESEARCH, 29, 3-4, 83, 87, HOKKAIDO UNIV, 1981年, ［査読有り］
英語, 研究論文（学術雑誌）

GROWTH OF AVIAN AND HUMAN INFLUENZA-VIRUSES IN ORGAN-CULTURES OF DUCK AND CHICKEN COLONS
C OHTA, R YANAGAWA, H KIDA
ARCHIVES OF VIROLOGY, 70, 2, 137, 145, SPRINGER-VERLAG WIEN, 1981年, ［査読有り］
英語, 研究論文（学術雑誌）

CLASSIFICATION OF AVIAN PARAMYXOVIRUSES BY IMMUNODIFFUSION ON THE BASIS OF ANTIGENIC SPECIFICITY OF THEIR M-PROTEIN ANTIGENS
H KIDA, R YANAGAWA
JOURNAL OF GENERAL VIROLOGY, 52, JAN, 103, 111, SOC GENERAL MICROBIOLOGY, 1981年, ［査読有り］
英語, 研究論文（学術雑誌）

DUCK INFLUENZA LACKING EVIDENCE OF DISEASE SIGNS AND IMMUNE-RESPONSE
H KIDA, R YANAGAWA, Y MATSUOKA
INFECTION AND IMMUNITY, 30, 2, 547, 553, AMER SOC MICROBIOLOGY, 1980年, ［査読有り］
英語, 研究論文（学術雑誌）

霊長類モデルを用いたインフルエンザとCOVID-19のワクチン開発
伊藤靖, 石垣宏仁, 仲山美沙子, NGUYEN Thanh Cong, 北川善紀, 安井文彦, 小原道法, 石井健, 新開大史, 迫田義博, 喜田宏, 日本実験動物学会総会講演要旨集(Web), 69th, 2022年

Selecting and using the appropriate influenza vaccine for each individual
Toshiki Sekiya, Marumi Ohno, Naoki Nomura, Chimuka Handabile, Masashi Shingai, David C. Jackson, Lorena E. Brown, Hiroshi Kida, Viruses, 13, 6, 2021年06月
Despite seasonal influenza vaccines having been routinely used for many decades, influenza A virus continues to pose a global threat to humans, causing high morbidity and mortality each year. The effectiveness of the vaccine is largely dependent on how well matched the vaccine strains are with the circulating influenza virus strains. Furthermore, low vaccine efficacy in naïve populations such as young children, or in the elderly, who possess weakened immune systems, indicates that influenza vaccines need to be more personalized to provide broader community protection. Advances in both vaccine technologies and our understanding of influenza virus infection and immunity have led to the design of a variety of alternate vaccine strategies to extend population protection against influenza, some of which are now in use. In this review, we summarize the progress in the field of influenza vaccines, including the advantages and disadvantages of different strategies, and discuss future prospects. We also highlight some of the challenges to be faced in the ongoing effort to control influenza through vaccination., 書評論文,書評,文献紹介等

ヒトH1N1インフルエンザAウイルスは発育鶏卵の羊膜・尿膜に発現するα2,3結合シアル酸を含む硫酸化糖鎖分子をレセプターとして利用する
高瀬明, 高瀬明, 一宮智美, 岡松正敏, 木下貴明, 木下貴明, 小林大樹, 市居修, 市居修, 山本直樹, 山本直樹, 迫田義博, 迫田義博, 喜田宏, 川島博人, 山本一夫, 西原祥子, 西原祥子, 日本ウイルス学会学術集会プログラム・予稿集(Web), 68th, 2021年

2020年度冬季に東日本の野鳥から分離された高病原性鳥インフルエンザウイルスの性状
磯田典和, 磯田典和, 日尾野隆大, TWABELA Augustin T., BAZARRAGCHAA Enkhbold, KIEN Le Trung, LOC Huynh Tan, 小笠原浩平, 林裕貴, 渡辺有希子, 齊藤慶輔, 喜田宏, 迫田義博, 迫田義博, 日本獣医学会学術集会講演要旨集, 164th (CD-ROM), 2021年

インフルエンザウイルスライブラリーを活用したパンデミックインフルエンザ対策
野村直樹, 新開大史, 松野啓太, 関屋俊輝, 関屋俊輝, 大野円実, 磯田典和, 迫田義博, 喜田宏, 喜田宏, 日本獣医学会学術集会講演要旨集, 164th (CD-ROM), 2021年

ヒトH1インフルエンザAウイルスの発育鶏卵での増殖に寄与するレセプター構造の解明
高瀬明, 高瀬明, 一宮智美, 岡松正敏, 木下貴明, 木下貴明, 小林大樹, 市居修, 山本直樹, 山本直樹, 迫田義博, 迫田義博, 喜田宏, 川島博人, 山本一夫, 西原祥子, 西原祥子, 日本糖質学会年会要旨集, 40th, 2021年

豚コレラウイルスワクチン株感染による細胞死誘導と細胞変性効果
板倉友香里, 板倉友香里, 松野啓太, 松野啓太, 伊藤麻子, 藤本悠理, 田村友和, 亀山健一郎, 岡松正敏, NICOLAS Ruggli, NICOLAS Ruggli, 喜田宏, 喜田宏, 澤洋文, 澤洋文, 迫田義博, 迫田義博, 日本ウイルス学会学術集会プログラム・予稿集(Web), 67th, 2019年

バロキサビル マルボキシル　　　　　　　　　　　　　　　
河野 茂, 齋藤 玲子, 渡辺 彰, 菅谷 憲夫, 喜田 宏, 三鴨 廣繁, 感染症学雑誌, 93, 1, 64, 65, 2019年01月
(一社)日本感染症学会, 日本語

バロキサビル マルボキシル　　　　　　　　　　　　　　　
河野 茂, 齋藤 玲子, 渡辺 彰, 菅谷 憲夫, 喜田 宏, 三鴨 廣繁, 日本化学療法学会雑誌, 67, 1, 112, 113, 2019年01月
(公社)日本化学療法学会, 日本語

海外から持ち込まれた未加熱家きん肉等から分離された高病原性及び低病原性H7N9亜型鳥インフルエンザウイルスの性状解析
柴田明弘, 岡松正敏, 住吉理穂, 松野啓太, 松野啓太, WANG Zu‐Jyun, WANG Zu‐Jyun, 喜田宏, 喜田宏, 尾坂優之, 迫田義博, 迫田義博, 日本獣医学会学術集会講演要旨集, 161st, 355, 2018年08月21日
日本語

2014-2017のベトナムにおけるH5高病原性鳥インフルエンザ発生の時空間・リスク解析(Spatiotemporal and risk analysis of H5 highly pathogenic avian influenza occurrence in Vietnam during 2014-2017)　　　　　　　　　　　　　　　
Nguyen Thanh-Lam, Stevenson Mark, Firestone Simon, Sims Les, Chu Huy, Nguyen Long, Nguyen Tien, 磯田 典和, 松野 啓太, 岡松 正敏, 喜田 宏, 迫田 義博, 日本獣医学会学術集会講演要旨集, 161回, 357, 357, 2018年08月
(公社)日本獣医学会, 英語

鳥インフルエンザウイルス検出におけるAlere i influenza A&Bの評価(Evaluation of Alere i influenza A&B for the detection of avian influenza virus)　　　　　　　　　　　　　　　
Enkhbold Bazarragchaa, 岡松 正敏, Ulaankhuu Ankhanbaatar, 松野 啓太, 喜田 宏, 新井 宏育, 迫田 義博, 日本獣医学会学術集会講演要旨集, 161回, 357, 357, 2018年08月
(公社)日本獣医学会, 英語

不活化インフルエンザウイルス全粒子ワクチンの非臨床試験
新開大史, 新開大史, 野村直樹, 関屋俊輝, 関屋俊輝, 大野円実, 笹田万友美, 大東卓史, 鈴木紗織, 石垣宏仁, 伊藤靖, 小笠原一誠, 小笠原一誠, 喜田宏, 喜田宏, 喜田宏, 日本ワクチン学会学術集会プログラム・抄録集, 22nd, 99, 2018年
日本語

サルモデルを用いた季節性インフルエンザに対する不活化全粒子ワクチンの有効性評価
伊藤靖, 鈴木紗織, 石垣宏仁, 仲山美沙子, 新開大史, 新開大史, 大東卓史, 野村直樹, 喜田宏, 喜田宏, 喜田宏, 小笠原一誠, 小笠原一誠, 日本ワクチン学会学術集会プログラム・抄録集, 22nd, 99, 2018年
日本語

H5N1高病原性鳥インフルエンザウイルスのシアル酸非依存的な細胞侵入経路の解析　　　　　　　　　　　　　　　
梶原 直樹, 野村 奈美子, 宇梶 麻紗子, 貞任 大地, 山本 直樹, 小原 道法, 安井 文彦, 迫田 義博, 喜田 宏, 芝崎 太, 生命科学系学会合同年次大会, 2017年度, [2P, 0957], 2017年12月
生命科学系学会合同年次大会運営事務局, 日本語

H5N1高病原性鳥インフルエンザウイルスのシアル酸非依存的な細胞侵入経路の解析　　　　　　　　　　　　　　　
梶原 直樹, 野村 奈美子, 宇梶 麻紗子, 貞任 大地, 山本 直樹, 小原 道法, 安井 文彦, 迫田 義博, 喜田 宏, 芝崎 太, 生命科学系学会合同年次大会, 2017年度, [2P, 0957], 2017年12月
生命科学系学会合同年次大会運営事務局, 日本語

高病原性鳥インフルエンザウイルスの進化―H5ウイルスを中心に―
迫田義博, 迫田義博, 喜田宏, 喜田宏, 感染症, 47, 6, 189‐199, 199, 2017年11月20日
H5N1亜型の高病原性鳥インフルエンザウイルスが1996年に中国で出現し、1997年には鳥からヒトへの感染が初めて報告され、世界的な関心が高まった。それから20年経過したが、ニワトリからアヒル農場での対策が不徹底な諸外国の影響を受け、ウイルスはいまだにこれらの国々の農場で流行を繰り返している。ウイルスは環境中に漏れ出し、渡り鳥に感染することで大陸を越えて運ばれる事態となり、日本もその被害国の1つである。H5ウイルスはヒトからヒトへの感染は報告されていないが、鳥からヒトに直接感染し、現在も死亡者が報告されている。初発生以降、中国や東南アジアでニワトリやアヒルに対しワクチンが使用されているため、ワクチン接種群での見えない感染(不顕性感染)が続き、同時に抗原変異株が次々と出現した。H5亜型高病原性鳥インフルエンザウイルスの進化は、20年間途切れることなく、ワクチン接種下でウイルスがニワトリやアヒルの間で受け継がれたことが最大の要因である。このウイルスの抗原性と遺伝子の進化(変異)は、すべての鳥インフルエンザ対策をさらに難しくさせている。このH5亜型に加え、H7亜型高病原性鳥インフルエンザウイルスが中国で新たに出現し、ヒトへの感染も報告されている。ニワトリやアヒル、すなわち家禽における鳥インフルエンザ対策を徹底し、環境中に漏れ出るウイルスをなくすこと以外、動物と人の健康を守る手段はない。(著者抄録), アステラス製薬(株), 日本語

鳥インフルエンザとパンデミックインフルエンザの要
喜田 宏, 医薬ジャーナル, 53, 10, 111, 115, 2017年10月

2.3.4.4高病原性鳥インフルエンザウイルスに対する新規モノクローナル抗体を用いた免疫クロマトグラフィーキットによるH5ヘマグルチニンの迅速かつ広範囲検出(Rapid and broad detection of H5 hemagglutinin by an immunochromatographic kit using novel monoclonal antibody against a 2.3.4.4 highly pathogenic avian influenza virus)　　　　　　　　　　　　　　　
Nguyen Thanh-Lam, 中石 和成, 本島 桂子, 大河原 彩子, 日尾野 隆大, 松野 啓太, 岡松 正敏, 高田 礼人, 喜田 宏, 迫田 義博, 日本獣医学会学術集会講演要旨集, 160回, 384, 384, 2017年08月
(公社)日本獣医学会, 英語

Rapid and broad detection of H5 hemagglutinin by an immunochromatographic kit using novel monoclonal antibody against a 2.3.4.4 highly pathogenic avian influenza virus(和訳中)　　　　　　　　　　　　　　　
Nguyen Thanh-Lam, 中石 和成, 本島 桂子, 大河原 彩子, 日尾野 隆大, 松野 啓太, 岡松 正敏, 高田 礼人, 喜田 宏, 迫田 義博, 日本獣医学会学術集会講演要旨集, 160回, 384, 384, 2017年08月
(公社)日本獣医学会, 英語

インフルエンザ罹患患児経過中の監護者における特異的免疫反応経時推移
熊谷卓司, 中山哲夫, 奥野良信, 森川佐依子, 加瀬哲男, 纐纈律子, 吉井洋紀, 本條健太, 迫田義博, 喜田宏, 庵原俊昭, 日本小児感染症学会総会・学術集会プログラム・抄録集, 49th, 136, 2017年
日本語

ワクチン接種後の鶏におけるH5鳥インフルエンザウイルスの抗原多様性の選択(Selection of antigenic variants of H5 avian influenza viruses in vaccinated chickens)　　　　　　　　　　　　　　　
Nguyen Thanh-Lam, 西 達也, 七戸 新太郎, Chu Duc-Huy, 日尾野 隆大, 松野 啓太, 岡松 正敏, 喜田 宏, 迫田 義博, 日本獣医学会学術集会講演要旨集, 159回, 371, 371, 2016年08月
(公社)日本獣医学会, 英語

新たなインフルエンザウイルスの出現と季節性インフルエンザワクチン
喜田 宏, 32, 6, 57, 64, 2016年05月

鳥インフルエンザとパンデミックインフルエンザ
喜田 宏, 感染症 最新の治療 2016-2018, 10, 13, 2016年04月

鳥，パンデミックと季節性インフルエンザをどうする
喜田 宏, 養鶏の友, 648, 14, 19, 2016年02月
日本畜産振興会, 日本語

実地医療に役立つインフルエンザトピックス(第13回)人獣共通感染症とインフルエンザ
喜田 宏, インフルエンザ, 17, 1, 15, 21, 2016年01月
メディカルレビュー社, 日本語

インフルエンザ患児の家族内被暴露監護者における血清抗体ならびに細胞性免疫反応経時推移
熊谷卓司, 中山哲夫, 奥野良信, 加瀬哲男, 森川佐依子, 迫田義博, 迫田義博, 喜田宏, 管秀, 庵原俊昭, 日本ワクチン学会学術集会プログラム・抄録集, 20th, 83, 2016年
日本語

インフルエンザワクチンの将来展望 (特集 成人の予防接種はどうあるべきか? : 予防医療推進の観点から) -- (成人予防接種の注意点と将来展望)
喜田 宏, 診断と治療, 103, 11, 1500, 1505, 2015年11月
診断と治療社, 日本語

鳥，パンデミックと季節性インフルエンザをどうする
喜田 宏, 畜産技術, 727, 37-43, 2015年11月

インフルエンザワクチンの将来展望
喜田 宏, 診断と治療, 103, 11, 1500, 1505, 2015年11月

鳥インフルエンザの問題点 : A(H5N1)とA(H7N9)を中心に (創刊50周年記念号) -- (特集 インフルエンザの最新事情とその対策 : One Healthの観点から)
喜田 宏, 医薬ジャーナル, 51, 10, 151, 154, 2015年10月
医薬ジャーナル社, 日本語

鳥インフルエンザの問題点 ～A(H5N1)とA(H7N9)を中心に～
喜田 宏, 医薬ジャーナル, 51, 10, 2401, 2404, 2015年10月

近年アジア地域で分離された鳥インフルエンザウイルスのアヒル,スズメ,カラス,およびクマネズミに対する病原性
小笠原浩平, 岡松正敏, 日尾野隆大, 七戸新太郎, 栗林沙弥, 市川貴也, 西達也, CHU Duc‐Huy, 鈴木瑞穂, 遠藤真由美, 田中和之, 谷川力, 喜田宏, 喜田宏, 喜田宏, 迫田義博, 迫田義博, 迫田義博, 日本獣医学会学術集会講演要旨集, 158th, 333, 333, 2015年08月30日
(公社)日本獣医学会, 日本語

インフルエンザウイルスのフコシル化α2,3シアロ糖認識機構の解析
日尾野隆大, 岡松正敏, 五十嵐学, 五十嵐学, MCBRIDE Ryan, DE VIRES Robert, PAULSON James, 松野啓太, 松野啓太, 迫田義博, 迫田義博, 喜田宏, 喜田宏, 喜田宏, 日本獣医学会学術集会講演要旨集, 158th, 332, 2015年08月30日
日本語

2014年のベトナムにおける介入有りまたは介入なしの生鳥市場にて単離された鳥インフルエンザウイルスの性状解析(Characterization of avian influenza viruses isolated in live bird markets with and without intervention in Vietnam in 2014)　　　　　　　　　　　　　　　
Chu Duc-Huy, 岡松 正敏, 松野 啓太, 日尾野 隆大, 小笠原 浩平, 鈴木 瑞穂, Nguyen Thanh Lam, 喜田 宏, 迫田 義博, 日本獣医学会学術集会講演要旨集, 158回, 329, 329, 2015年08月
(公社)日本獣医学会, 英語

インフルエンザウイルスのフコシル化α2,3シアロ糖認識機構の解析
日尾野隆大, 岡松正敏, 五十嵐学, 五十嵐学, MCBRIDE Ryan, DE VIRES Robert P, PAULSON James C, 迫田義博, 迫田義博, 喜田宏, 喜田宏, 日本糖質学会年会要旨集, 34th, 173, 2015年07月01日
日本語

鳥インフルエンザウイルスはパンデミックを起こすか？
喜田 宏, 臨床と微生物, 42, 3, 249, 254, 2015年05月

全粒子ワクチンは複数のクレードのH5N1高病原性鳥インフルエンザウイルスに対する記憶免疫反応を誘導する　　　　　　　　　　　　　　　
仲山 美沙子, 七戸 新太郎, 伊藤 靖, 石垣 宏仁, 北野 光崇, 有方 雅彦, Pham Van Loi, 石田 英晃, 北川 直子, 小笠原 一誠, 土屋 英明, 中村 紳一朗, 岡松 正敏, 迫田 義博, 市川 貴也, 喜田 宏, Le Quynh Mai, 伊藤 睦美, 河岡 義裕, 滋賀医学, 37, 146, 146, 2015年03月
(一社)滋賀県医師会, 日本語

ヒトH1インフルエンザウイルスは6硫酸化糖鎖構造をレセプターとするか?
一宮智美, 高瀬明, 木下聖子, 喜田宏, 西原祥子, 日本糖質学会年会要旨集, 34th, 2015年

北海道大学人獣共通感染症リサーチセンターザンビア拠点における感染症対策への取り組み　　　　　　　　　　　　　　　
小川寛人, 喜田宏, 最新医学, 2015年

季節性インフルエンザワクチンの現状と展望
喜田 宏, 化学療法の領域, 30, 12, 84, 89, 2014年12月

日本および世界各地における高病原性および低病原性鳥インフルエンザウイルスの家禽および野鳥における感染状況
岡松 正敏, 迫田 義博, 喜田 宏, 化学療法の領域, 30, 12, 25, 32, 2014年12月

インフルエンザの最新知見~鳥,パンデミックと季節性インフルエンザ対策をどうするか~1.日本および世界各地における高病原性および低病原性鳥インフルエンザウイルスの家禽および野鳥における感染状況
岡松正敏, 迫田義博, 喜田宏, 化学療法の領域, 30, 12, 2173, 2180, 2014年11月25日
日本語

豚コレラウイルス非構造蛋白NS4BのN末端両親媒性ヘリックスは豚に対する病原性に関与する
田村友和, RUGGLI Nicolas, LINGER Matthias, 長島尚史, 峯淳貴, 岡松正敏, HOFMANN Martin A, SUMMERFIELD Artur, 喜田宏, 迫田義博, 日本ウイルス学会学術集会プログラム・抄録集, 62nd, 250, 2014年10月31日
日本語

鳥インフルエンザウイルスのヒト型レセプター特異性獲得メカニズムの解析
柳田里澄, 岡松正敏, 日尾野隆大, 迫田義博, 喜田宏, 日本ウイルス学会学術集会プログラム・抄録集, 62nd, 155, 2014年10月31日
日本語

H5N1高病原性鳥インフルエンザ組換えワクシニアワクチン単回接種による免疫長期持続作用機序の解析
安井文彦, 宗片圭祐, 倉石武, 服部正策, 藤幸知子, 米田美佐子, 迫田義博, 喜田宏, 甲斐知恵子, 小原道法, 日本ウイルス学会学術集会プログラム・抄録集, 62nd, 235, 2014年10月31日
日本語

H5N1高病原性鳥インフルエンザウイルスがブタに感染するとSAα2,6Galレセプターに結合するウイルスが選択されるか?
七戸新太郎, 迫田義博, 西達也, 日尾野隆大, 岡松正敏, 伊藤靖, 小笠原一誠, 喜田宏, 日本ウイルス学会学術集会プログラム・抄録集, 62nd, 154, 2014年10月31日
日本語

2010年から2014年に家禽及び野鳥から分離されたH5及びH7鳥インフルエンザウイルスの遺伝子と抗原性の解析
迫田義博, 大河原彩子, 日尾野隆大, 小笠原浩平, 岡松正敏, 喜田宏, 日本ウイルス学会学術集会プログラム・抄録集, 62nd, 163, 2014年10月31日
日本語

ニワトリのインフルエンザウイルスはフコシル化および硫酸化α2,3シアル酸糖鎖をレセプターとする
岡松正敏, 日尾野隆大, 五十嵐学, DE VRIES Robert, 西原祥子, 高瀬明, PAULSON James C, 迫田義博, 喜田宏, 日本ウイルス学会学術集会プログラム・抄録集, 62nd, 154, 2014年10月31日
日本語

H5N1インフルエンザウイルスに対する中和抗体を用いた受動免疫の感染防御効果
吉田玲子, 伊藤靖, 七戸新太郎, 小笠原一誠, 日尾野隆大, 岡松正敏, 迫田義博, 喜田宏, LE Mai Quynh, 河岡義裕, 五十嵐学, 鈴木定彦, 高田礼人, 日本ウイルス学会学術集会プログラム・抄録集, 62nd, 234, 2014年10月31日
日本語

Research activities of Hokudai center for Zoonosis Control in Zambia
Hideaki Higashi, Hiroshi Kida, Journal of Disaster Research, 9, 5, 818, 822, 2014年10月01日
The Hokkaido University Research Center for Zoonosis Control (CZC) established the Hokudai Center for Zoonosis Control in Zambia (HUCZCZ) at the School of Veterinary Medicine, the University of Zambia, in 2007 to control zoonotic diseases in the areas of South Africa, where various emerging infectious diseases have occurred. The CZC promotes epidemiological studies and basic researches of infectious diseases caused by viruses, protozoa, and bacteria by using the biosafety level 3 facility in the HUCZCZ. This article introduces research activities of the HUCZCZ in Zambia., Fuji Technology Press, 英語, 書評論文,書評,文献紹介等

鳥インフルエンザの現状と今後
喜田 宏, 医薬ジャーナル, 50, 10, 107, 112, 2014年10月

牛ウイルス性下痢ウイルス準種のcDNAクローン構築と自然免疫制御能の解析
塩川舞, 加藤あやの, 藤本悠理, 安部優里, 田村友和, 西根薫, 峯淳貴, 青木博史, 福所秋雄, 喜田宏, 迫田義博, 日本獣医学会学術集会講演要旨集, 157th, 396, 396, 2014年08月11日
(公社)日本獣医学会, 日本語

2010年から2013年に家禽及び野鳥から分離されたH5及びH7鳥インフルエンザウイルスの遺伝子と抗原性の解析
大河原彩子, 日尾野隆大, 小笠原浩平, 岡松正敏, 迫田義博, 喜田宏, 日本獣医学会学術集会講演要旨集, 157th, 411, 2014年08月11日
日本語

糖鎖アレイを用いたカモとニワトリのインフルエンザウイルスのレセプター特異性
岡松正敏, DE VRIES Robert, 日尾野隆大, 迫田義博, PAULSON C James, 喜田宏, 日本獣医学会学術集会講演要旨集, 157th, 419, 2014年08月11日
日本語

豚コレラウイルス非構造蛋白NS4BのN末端領域は豚に対する病原性に関与する
田村友和, RUGGLI Nicolas, 長島尚史, 峯淳貴, 岡松正敏, SUMMERFIELD Artur, HOFMANN Martin A, 喜田宏, 迫田義博, 日本獣医学会学術集会講演要旨集, 157th, 410, 2014年08月11日
日本語

ニワトリのインフルエンザウイルスはフコシル化α2,3シアル酸糖鎖をレセプターとする
日尾野隆大, 岡松正敏, 五十嵐学, 西原祥子, 高瀬明, 迫田義博, 喜田宏, 日本糖質学会年会要旨集, 33rd, 62, 2014年07月23日
日本語

不活化全粒子インフルエンザワクチンを内包したマイクロニードルアレイによる免疫誘導特性の評価
小山田孝嘉, 来馬浩二, 島田俊雄, 迫田義博, 喜田宏, 日本DDS学会学術集会プログラム予稿集, 30th, 184, 2014年07月01日
日本語

ワクチン研究者に聞く
喜田 宏, ワクチンジャーナル, 2, 1, 40, 44, 2014年06月

感染症研究国際ネットワーク研究の現状 7.ザンビア‐北海道大学人獣共通感染症リサーチセンターザンビア拠点
東秀明, 東秀明, 喜田宏, 化学療法の領域, 30, 6, 1234, 1239, 2014年05月25日
日本語

私のインフルエンザ研究
喜田宏, 喜田宏, 喜田宏, 喜田宏, 臨床とウイルス, 42, 2, S6-S7, 2014年05月01日
日本語

次世代型感染症ワクチン 今,インフルエンザワクチンに何が求められるか
喜田宏, 喜田宏, 喜田宏, 喜田宏, 喜田宏, 最新医学, 69, 4, 858, 864, 2014年04月10日
最新医学社, 日本語

カニクイザルにおける2013年H7N9亜型インフルエンザウイルスに対するワクチンとNA阻害剤の効果
伊藤靖, 仲山美沙子, 石垣宏仁, 石田英晃, 七戸新太郎, 岡松正敏, 迫田義博, 喜田宏, 喜田宏, 小笠原一誠, 日本病理学会会誌, 103, 1, 224, 2014年03月26日
日本語

免疫低下状態のカニクイザルにおけるH1N1亜型インフルエンザウイルスの病原性の解析
伊藤靖, LOI Pham Van, 仲山美沙子, 石垣宏仁, 有方雅彦, 石田英晃, 北川直子, 小笠原一誠, 七戸新太郎, 岡松正敏, 迫田義博, 喜田宏, 喜田宏, 滋賀医学, 36, 142, 2014年03月20日
日本語

どうする?鳥インフルエンザとパンデミックインフルエンザ
喜田宏, 喜田宏, 日本環境感染学会誌, 29, Supplement, 137, 2014年01月24日
日本語

鳥インフルエンザウイルスの病原性と感染対策
喜田宏, 喜田宏, 感染症, 44, 1, 13-18,9, 2014年01月20日
日本語

豚コレラウイルスALD/A76株の病原性解析に用いる完全長cDNAクローンおよびレプリコンの作出
藤本悠理, 田村友和, 遠藤真由美, 吉野史, RUGGLI Nicolas, 喜田宏, 喜田宏, 迫田義博, 日本獣医学会学術集会講演要旨集, 157th, 2014年

鳥インフルエンザ
喜田宏, 喜田宏, 喜田宏, 喜田宏, 感染症内科, 1, 6, 564, 572, 2013年12月28日
日本語

グローバル化する感染症の最新情報 ウイルス感染症 インフルエンザ
喜田宏, 月刊臨床と研究, 90, 12, 1666, 1673, 2013年12月20日
大道学館出版部, 日本語

微生物ABC インフルエンザウイルス
喜田宏, Up-to-date子どもの感染症, 1, 2, 74, 81, 2013年12月10日
メディカルレビュー社, 日本語

人獣共通感染症としてのインフルエンザ
喜田宏, 月刊養豚界, 48, 11, 20, 24, 2013年11月01日
緑書房, 日本語

クローズアップ インフルエンザ インフルエンザウイルスの生態
喜田宏, 小児内科, 45, 11, 1927, 1934, 2013年11月01日
日本語

GANPトランスジェニックマウスを用いたH5N1高病原性鳥インフルエンザウイルスHAに対する汎クレード高親和性中和単クローン抗体の開発
安井文彦, 桑原一彦, 飛田良美, 棟方翼, 斉藤誠, 七戸新太郎, 迫田義博, 喜田宏, 阪口薫雄, 小原道法, 日本ウイルス学会学術集会プログラム・抄録集, 61st, 153, 2013年10月29日
日本語

糖鎖アレイデータベースから抽出されたインフルエンザウイルス感染を増強する糖鎖構造
一宮智美, 西原祥子, 木下聖子, 山本直樹, 喜田宏, 高瀬明, 日本ウイルス学会学術集会プログラム・抄録集, 61st, 239, 2013年10月29日
日本語

2010年に稚内で分離された2株の高病原性鳥インフルエンザウイルスの病原性比較
直亨則, 梶原将大, 丸山隼輝, 木村享史, MANZOOR Rashid, 村松美笑子, 岡松正敏, 宮本洋子, 吉田玲子, 迫田義博, 喜田宏, 高田礼人, 日本ウイルス学会学術集会プログラム・抄録集, 61st, 150, 2013年10月29日
日本語

高病原性鳥インフルエンザ抗原タンパク質発現組換え麻疹ウイルスのサル感染モデルにおける防御効果の検討
藤幸知子, 米田美佐子, 堀江亮, 倉石武, 池田房子, 石井美穂, 小見美央, 安井文彦, 宗片圭祐, 佐藤宏樹, 服部正策, 喜田宏, 小原道法, 甲斐知恵子, 日本ウイルス学会学術集会プログラム・抄録集, 61st, 155, 2013年10月29日
日本語

計算科学手法を用いたインフルエンザウイルスヘマグルチニンと亜型間交差反応性抗体との分子間相互作用の解析
五十嵐学, 吉田玲子, 関嶋政和, 常盤広明, 喜田宏, 伊藤公人, 高田礼人, 日本ウイルス学会学術集会プログラム・抄録集, 61st, 239, 2013年10月29日
日本語

ウイルス学の立場から―鳥インフルエンザとパンデミックインフルエンザ対策の基本―
喜田宏, 感染症道場, 2, 4, 59, 61, 2013年10月20日
日本語

Influenza virus-like particles containing HA, NA, and M1 induced protection in chickens against a lethal challenge with the highly pathogenic H5N1 avian influenza virus
Motofumi Shimizu, Satomi Yanase, O. O. Chang Myint, Masatoshi Okamatsu, Yoshihiro Sakoda, Hiroshi Kida, Hiroshi Takaku, Hiroshi Takaku, Journal of Vaccines and Vaccination, 4, 2013年10月04日
Background: Intermittent outbreaks of highly pathogenic avian influenza virus (flu) infections illustrate the potential for pandemic spread of this deadly disease, thus making the development of sufficient supplies of safe vaccines a necessity. Influenza virus-like particles (Vlps) have been suggested as a promising non-egg or non-mammalian cell culture-based candidate for vaccination against flu. Vlps containing hemagglutinin have previously been shown to promote protection against homologous viral strains. In this report, we describe the development of an H5N1 flu Vlp vaccine involving only three flu viral structural proteins, (i.e., HA, NA, and M1), which were derived from an avian flu A/duck/Hokkaido/vac-1/2004 (H5N1) virus. The H5N1 Vlps produced from insect cells exhibited hemagglutination and neuraminidase activities and generated an immune response in BALB/c mice. We additionally performed viral challenge studies using chickens. Methodology and results: Vlps consisting of the hemagglutinin (HA), neuraminidase (NA), and matrix 1 (M1) proteins of A/duck/Hokkaido/vac-1/2004 (H5N1) were transferred using baculovirus within Spodoptera frugiperda (Sf9) cells. Mice were first immunized with Vlps, and the immune response was compared between animals vaccinated with HA and NA-HA-negative M1 Vlps. The IgG levels of HA-M1 Vlp- and NA-M1 Vlp-treated groups were observed, and 5-fold higher levels of H5N1-specific antibodies were induced in the groups of mice immunized with HA-NA-M1 Vlp. The HA-NA and HA-NA-M1 Vlps vaccines induced IgG2a and IgG2b antibodies as well as IgG1 antibodies, indicating that both Th1 and Th2 immune responses were induced. Furthermore, NA-M1 immunization induced IgG and IgG1 isotype antibodies and led to low levels of IgG2a and IgG2b. Additionally, all chickens immunized with HA-NA-M1 Vlp were protected against deadly infections with the highly pathogenic avian flu virus A/chicken/Yamaguchi/4/2004 (H5N1). Conclusions: Intramuscular administration of H5 Vlps conferred immunity against a deadly viral challenge. The H5 Vlp vaccine was more successful at raising Th1-biased protective responses, including IgG2a production. Thus, flu Vlps offer an important approach for immunization, especially if a pandemic occurs. Recognizing the current state of vaccination strategies, it is imperative to investigate the associated immunogenicity and defensive capabilities of H5 Vlps in comparison with inactivated whole virus and attenuated live H5N1 viruses. These results support the continued development of Vlps as a formulation for a vaccine against flu infection. © 2013 Shimizu M, et al.

これからのインフルエンザ診療―的確な診断・適正な治療・先手の予防 H7N9およびH5N1鳥インフルエンザウイルスについて
喜田宏, 感染と抗菌薬, 16, 3, 272, 277, 2013年09月10日
ヴァンメディカル, 日本語

H7N9およびH5N1鳥インフルエンザウイルスについて (特集 これからのインフルエンザ診療 : 的確な診断・適正な治療・先手の予防)
喜田 宏, 感染と抗菌薬, 16, 3, 272, 277, 2013年09月
ヴァンメディカル, 日本語

抗PB2細胞内発現単クローン抗体のA型インフルエンザウイルスに対する感染抵抗性
藤本佳万, 尾崎絹代, 前田雅弘, 西島謙一, 高桑弘樹, 大槻公一, 喜田宏, 小野悦郎, 日本獣医学会学術集会講演要旨集, 156th, 270, 2013年08月30日
日本語

牛ウイルス性下痢ウイルスのcDNAクローンの作出と自然免疫抑制機能の解析
安部優里, 迫田義博, 三津橋和也, 田村友和, 小佐々隆志, 中村成幸, 喜田宏, 日本獣医学会学術集会講演要旨集, 156th, 265, 2013年08月30日
日本語

H7低病原性鳥インフルエンザウイルスのニワトリに対する病原性獲得メカニズム
市川貴也, 岡松正敏, 佐藤由佳, 日尾野隆大, 迫田義博, 喜田宏, 日本獣医学会学術集会講演要旨集, 156th, 269, 2013年08月30日
日本語

鳥インフルエンザの正体とその克服法 (研究小集会(第9回)卵部会)
喜田 宏, 日本食品科学工学会誌 = Journal of the Japanese Society for Food Science and Technology, 60, 7, 371, 374, 2013年07月
日本食品科学工学会, 日本語

鳥インフルエンザの現状と対策
岡松正敏, 喜田宏, 日本獣医師会雑誌, 66, 6, 352, 354, 2013年06月20日
日本獣医師会, 日本語

ニワトリのインフルエンザウイルスはニワトリ気管上皮に発現するフコシル化α2,3シアル酸糖鎖をレセプターとして認識する
日尾野隆大, 岡松正敏, 西原祥子, 高瀬明, 迫田義博, 喜田宏, 日本獣医学会学術集会講演要旨集, 155th, 229, 2013年03月04日
日本語

鳥インフルエンザウイルスがブタに感染するとSAα2,6Galレセプターを認識するウイルスが選択される
七戸新太郎, 岡松正敏, 迫田義博, 喜田宏, 日本獣医学会学術集会講演要旨集, 155th, 229, 2013年03月04日
日本語

タイで分離されたEND陰性豚コレラウイルスの遺伝子と病原性の解析
迫田義博, 三津橋和也, 田村友和, 長島尚史, 岡松正敏, NICOLAS Ruggli, SUGIRA Parchariyanon, WASANA Pinyochon, 喜田宏, 日本獣医学会学術集会講演要旨集, 155th, 226, 2013年03月04日
日本語

高病原性鳥インフルエンザとパンデミックインフルエンザ
喜田宏, 感染症道場, 2, 1, 30, 33, 2013年01月20日
メディカルレビュー社, 日本語

ワクチニアウイルスDIs株を母体としたインフルエンザHA組換えワクチンの混合接種によるカニクイザルでの発症防御効果
宗片圭祐, 安井文彦, 伊藤靖, 石井孝司, 七戸新太郎, 喜田宏, 小笠原一誠, 小原道法, 日本ワクチン学会学術集会プログラム・抄録集, 17th, 160, 2013年
日本語

カニクイザルでのH5N1高病原性トリインフルエンザ組換えワクシニアワクチンの単回接種による長期防御効果の検討
安井文彦, 宗片圭祐, 倉石武, 服部正策, 藤幸知子, 米田美佐子, 迫田義博, 喜田宏, 甲斐知恵子, 小原道法, 日本ワクチン学会学術集会プログラム・抄録集, 17th, 87, 2013年
日本語

急性呼吸器感染症の感染メカニズムと疫学,感染予防・制御に関する研究 鳥,ブタなどのインフルエンザ疫学と感染制御体制の検討
喜田宏, 急性呼吸器感染症の感染メカニズムと疫学、感染予防・制御に関する研究 平成24年度 総括・分担研究報告書, 77, 81, 2013年
日本語

蛍光イムノクロマトグラフィ法を用いた高病原性トリインフルエンザウイルスH5亜型の広域検出法の確立
櫻井陽, 高山勝好, 野村奈美子, 迫田義博, 須田美彦, 小林行治, 阪口薫雄, 喜田宏, 小原道法, 芝崎太, 日本分子生物学会年会プログラム・要旨集(Web), 36th, WEB ONLY 3P-1007, 2013年
日本語

微生物と感染症診療 高病原性鳥インフルエンザとパンデミックインフルエンザ
喜田 宏, 感染症道場, 2, 1, 30, 33, 2013年01月
メディカルレビュー社, 日本語

鳥インフルエンザの正体とその克服法
喜田 宏, 日本食品科学工学会誌, 60, 7, 371, 374, 2013年
Highly pathogenic avian influenza virus (HPAIV) causes lethal infection in gallinaceous poultry such as chickens. HPAIV is generated when a nonpathogenic virus brought in by migratory birds from nesting lakes in the north is transmitted to chickens via domestic ducks, geese, quails, etc. and acquires pathogenicity for chickens with repeated multiple infections in the chicken population. Now H5N1 HPAIV has spread to 62 countries in Eurasia and Africa. H5N1 HPAIVs were isolated from dead water birds in Mongolia and Hokkaido, Japan on the way back to their nesting lakes in Siberia in April to May 2005, 2006, 2008, 2009 and 2010. It is concerned that these HPAI viruses may perpetuate in the lakes where migratory water birds nest in summer. On 14<SUP>th</SUP> October in 2010, 2 H5N1 HPAIVs were isolated from fecal samples of ducks who flew from Siberia to Ohnuma Lake in Wakkanai, Hokkaido, Japan. Since then, 24 outbreaks of avian influenza due to infection with the H5N1 viruses closely related to the viruses isolated from fecal samples of ducks in Hokkaido in October 2010 have occurred in 9 different prefectures in Japan until the end of March 2011. The other serious concern is that 620 people have been infected with the H5N1 virus, 60% of whom died in 15 countries since 2004 (as of 15 February 2013). It is noted that most of the human cases (86%) are in China, Viet Nam, Indonesia, and Egypt where bird flu vaccines are used. It is strongly proposed to eradicate the H5N1 HPAIVs from Asia by stamping-out without misuse of vaccine through international collaboration under the umbrella of "One World, One Health" concept., 社団法人 日本食品科学工学会, 日本語

パンデミックインフルエンザの発生に即応した,ワクチン製造用種ウイルス株の作出―インフルエンザウイルスライブラリーの利用―
岡本成史, 竹中延之, 迫田義博, 岡松正敏, 山本直樹, ハリディ アーマド, 山田博司, 森康子, 喜田宏, 山西弘一, 日本ウイルス学会学術集会プログラム・抄録集, 60th, 263, 2012年10月31日
日本語

鳥インフルエンザウイルスがブタに感染するとSAα2,6Galレセプターを認識するウイルスが選択される
七戸新太郎, 岡松正敏, 迫田義博, 喜田宏, 日本ウイルス学会学術集会プログラム・抄録集, 60th, 167, 2012年10月31日
日本語

豚コレラウイルス非構造蛋白N^proのアミノ酸を置換したウイルスの感染に対する豚の自然免疫応答と病原性の解析
田村友和, 長島尚史, 山本直樹, 岡松正敏, NICOLAS Ruggli, 迫田義博, 喜田宏, 日本ウイルス学会学術集会プログラム・抄録集, 60th, 250, 2012年10月31日
日本語

H3N2インフルエンザウイルスHAへの糖鎖付加と周辺アミノ酸残基にかかる選択圧の経詩的変化
五十嵐学, 中尾亮, 喜田宏, 高田礼人, 伊藤公人, 日本ウイルス学会学術集会プログラム・抄録集, 60th, 318, 2012年10月31日
日本語

渡り鳥とインフルエンザ
喜田 宏, 科学, 82, 8, 918, 922, 2012年08月
岩波書店, 日本語

グルタルアルデヒドを主成分とする消毒薬の鳥インフルエンザウイルスに対する不活化効果
迫田 義博, 遠藤 真由美, 佐藤 由佳, 岡松 正敏, 喜田 宏, 日本獣医師会雑誌 = Journal of the Japan Veterinary Medical Association, 65, 4, 303, 305, 2012年04月20日
グルタルアルデヒドを主成分とする消毒薬の鳥インフルエンザウイルスに対する不活化効果を調べた．その結果，グルタルアルデヒドを主成分とする消毒薬は有機物が混入しても高い消毒効果が認められた．しかし，低温下ではその効果は低下した．またウイルスの不活化には，塩化ジデシルジメチルアンモニウムを主成分とする陽イオン系界面活性剤に比べ，作用時間をより長く必要とすることがわかった．以上の結果は，グルタルアルデヒドを主成分とする消毒薬を鳥インフルエンザウイルスに対して用いる場合には，有機物の有無に関わらずその効果が期待できるが，反応温度や反応時間を十分に考慮する必要があることを示している．, 日本獣医師会, 日本語

豚インフルエンザ総説 (特集 インフルエンザから豚を守ろう)
喜田 宏, 七戸 新太郎, 養豚の友, 0, 515, 14, 16, 2012年02月
日本畜産振興会, 日本語

Galectin-9は細胞およびマウスにおけるインフルエンザAウイルスの感染を抑制する
服部ともえ, 服部ともえ, 丸山隼輝, 藤岡容一朗, 中山洋佑, 大場雄介, 仁木敏郎, 有川智博, 宮崎忠昭, 平島光臣, 喜田宏, 喜田宏, 日本ウイルス学会学術集会プログラム・抄録集, 60th, 2012年

豚コレラウイルスの豚における増殖とサイトカイン応答
長島尚史, 迫田義博, 田村友和, 山本直樹, 岡松正敏, NICOLAS Ruggli, 喜田宏, 喜田宏, 日本ウイルス学会学術集会プログラム・抄録集, 60th, 2012年

豚コレラウイルスの病原性に関与するアミノ酸置換が宿主のサイトカイン応答に及ぼす影響
長島尚史, 迫田義博, 田村友和, 栗林沙弥, 山本直樹, 岡松正敏, RUGGLI Nicolas, 喜田宏, 喜田宏, 日本獣医学会学術集会講演要旨集, 153rd, 2012年

北海道で分離された牛ウイルス性下痢ウイルス1aおよび1b亜型株の遺伝子と抗原性の解析
安部優里, 迫田義博, 三津橋和也, 田村友和, 長島尚史, 岡松正敏, 長井誠, 喜田宏, 喜田宏, 日本獣医学会学術集会講演要旨集, 154th, 2012年

対談 鳥インフルエンザの諸問題
柏木 征三郎, 喜田 宏, インフルエンザ, 13, 1, 9, 19, 2012年01月
メディカルレビュー社, 日本語

序論 : 人獣共通感染症の克服戦略 (特集 人獣共通感染症の克服に向けて)
喜田 宏, 最新医学, 66, 12, 2609, 2611, 2011年12月
最新医学社, 日本語

対談 人獣共通感染症の克服対策 (特集 人獣共通感染症の克服に向けて)
倉田 毅, 喜田 宏, 最新医学, 66, 12, 2612, 2622, 2011年12月
最新医学社, 日本語

インフルエンザ (特集 人獣共通感染症の克服に向けて)
喜田 宏, 最新医学, 66, 12, 2623, 2631, 2011年12月
最新医学社, 日本語

高病原性鳥インフルエンザと対策(3・最終回)鳥とヒトのインフルエンザ対策はいかにあるべきか
喜田 宏, 鶏の研究, 86, 12, 22, 25, 2011年12月
木香書房, 日本語

高病原性鳥インフルエンザと対策(2)H5N1高病原性インフルエンザウイルスが常在化したのはなぜか
喜田 宏, 鶏の研究, 86, 11, 14, 17, 2011年11月
木香書房, 日本語

高病原性鳥インフルエンザと対策(1)高病原性インフルエンザ発生の現状
喜田 宏, 鶏の研究, 86, 10, 14, 17, 2011年10月
木香書房, 日本語

2009年のウズラ由来H7N6亜型高病原性鳥インフルエンザウイルスの感染経路の推定　　　　　　　　　　　　　　　
内田 裕子, 金平 克史, 真瀬 昌司, 竹前 喜洋, 渡辺 千晶, 笛吹 達史, 藤本 佳万, 伊藤 壽啓, 五十嵐 学, 伊藤 公人, 高田 礼人, 迫田 義博, 岡松 正敏, 山本 祐, 中村 菊保, 喜田 宏, 廣本 靖明, 津田 知幸, 西藤 岳彦, 動物衛生研究成果情報, 10, 15, 16, 2011年09月
(国研)農業・食品産業技術総合研究機構動物衛生研究所, 日本語

高病原性鳥インフルエンザの畜産と公衆衛生に与えるインパクト--ワクチン頼みから摘発・淘汰に転換することがウイルスを一掃する (特集 冬場に向けた衛生対策--畜種別の留意点と感染症予防対策・畜舎環境改善)
喜田 宏, 畜産コンサルタント, 47, 9, 24, 29, 2011年09月
中央畜産会, 日本語

人獣共通感染症リサーチセンター・喜田宏氏の講演より インフルエンザウイルスの生態を知る
喜田 宏, 養鶏の友, 595, 34, 41, 2011年09月
日本畜産振興会, 日本語

2010‐2011年に日本で野鳥から分離されたH5N1高病原性鳥インフルエンザウイルス
岡松正敏, 伊藤啓史, 内田裕子, 迫田義博, 山本直樹, 曽田公輔, 笛吹達史, 尾崎弘一, 山口剛士, 村瀬敏之, 伊藤壽啓, 西藤岳彦, 喜田宏, 日本獣医学会学術集会講演要旨集, 152nd, 231, 2011年08月31日
日本語

高病原性鳥インフルエンザの防疫--迅速診断、摘発・淘汰が基本
喜田 宏, 鶏の研究, 86, 6, 52, 54, 2011年06月
木香書房, 日本語

鳥インフル克服に向け抜本対策を (特集 「家畜災害」克服への検証)
喜田 宏, AFCフォーラム, 58, 13, 3, 6, 2011年04月
日本政策金融公庫農林水産事業本部, 日本語

高病原性鳥インフルエンザの防疫 (特集 高病原性鳥インフルエンザの対策と実態)
喜田 宏, 畜産技術, 0, 671, 2, 6, 2011年04月
畜産技術協会, 日本語

豚コレラウイルス非構造蛋白N^proへのアミノ酸変異導入による自然免疫の抑制と豚に対する病原性の上昇
迫田義博, 田村友和, 長島尚史, 山本直樹, 岡松正敏, RUGGLI Nicolas, 喜田宏, 喜田宏, 日本獣医学会学術集会講演要旨集, 152nd, 2011年

豚コレラ弱毒生ワクチン株の豚継代による病原性獲得に与る因子の検索
田村友和, 迫田義博, 吉野史, 長島尚史, 山本直樹, 佐藤由佳, 岡松正敏, RUGGLI Nicolas, 喜田宏, 喜田宏, 日本獣医学会学術集会講演要旨集, 152nd, 2011年

牛ウイルス性下痢ウイルス2b亜型(BVDV‐2b)株の性状
田村友和, 迫田義博, 吉野史, 杉田征彦, 岡松正敏, 長井誠, 伊藤美加, 青木博史, 喜田宏, 日本ウイルス学会学術集会プログラム・抄録集, 58th, 275, 2010年10月15日
日本語

H3N2インフルエンザウイルスHAに付加された糖鎖は周辺エピトープを覆い隠していたか?〜糖鎖付加部位周辺アミノ酸残基の多様性変化の解析〜
五十嵐学, 高田礼人, 喜田宏, 喜田宏, 喜田宏, 伊藤公人, 伊藤公人, 日本ウイルス学会学術集会プログラム・抄録集, 58th, 227, 2010年10月15日
日本語

パンデミックインフルエンザAウイルス(H1N1)ヘマグルチニンに対するモノクローナル抗体の性状解析
吉田玲子, 苫米地大輔, 五十嵐学, 宮本洋子, 加瀬哲男, 喜田宏, 喜田宏, 喜田宏, 高田礼人, 日本ウイルス学会学術集会プログラム・抄録集, 58th, 219, 2010年10月15日
日本語

牛ウイルス性下痢ウイルス2b亜型(BVDV‐2b)株の性状
田村友和, 迫田義博, 吉野史, 岡松正敏, 長井誠, 伊藤美加, 青木博史, 喜田宏, 日本獣医学会学術集会講演要旨集, 150th, 234, 234, 2010年09月01日
(公社)日本獣医学会, 日本語

パンデミックの出現に関する展望と今後の対策 (特集 検証「パンデミックインフルエンザ2009」)
喜田 宏, 公衆衛生, 74, 8, 687, 690, 2010年08月
医学書院, 日本語

鳥, ブタ, そしてパンデミックインフルエンザ騒動を振り返る
喜田 宏, ウイルス, 60, 1, 17, 20, 2010年06月01日
日本ウィルス学会, 日本語

Intravenous peramivir for treatment of influenza A and B infection in high-risk patients
S. Kohno, H. Kida, M. Mizuguchi, N. Hirotsu, T. Ishida, J. Kadota, J. Shimada, INTERNATIONAL JOURNAL OF INFECTIOUS DISEASES, 14, E85, E86, 2010年03月
ELSEVIER SCI LTD, 英語, 研究発表ペーパー・要旨（国際会議）

Epidemiology of avian influenza viruses in wild birds in Mongolia
E. -O. Tseren-Ochir, B. Damdinjav, T. Sharkhuu, H. M. Kang, Y. Sakoda, B. Purevsuren, S. Ruuragchaa, Y. J. Lee, H. Kida, B. Khishgee, S. Sengee, INTERNATIONAL JOURNAL OF INFECTIOUS DISEASES, 14, E164, E165, 2010年03月
ELSEVIER SCI LTD, 英語, 研究発表ペーパー・要旨（国際会議）

P-3 ハウアミンBの合成研究(ポスター発表の部)
田中 剛, 乾 広美, 紀田 博志, 児玉 猛, 岡本 拓也, 竹島 亜季, 舘 祥光, 森本 善樹, 天然有機化合物討論会講演要旨集, 52, 0, 247, 252, 2010年
Haouamines A (1) and B (2) were isolated from the marine ascidian Aplidium haouarianum collected off the coast of southern Spain. They belong to an unprecedented class of alkdaloids. A unique feature of haouamines is an eleven-membered 3-aza-[7]-paracyclophane moiety, which is so highly strained. The B ring of paracyclophane moiety is non planar and exists in a pseudo-boat conformation. Other features are a diaryl-substituted stereogenic quaternary center and an anti-Bredt double bond. In solution, each haouamine exists as an inseparable mixture of two interconverting isomers. Haouamine A exibits cytotoxic activity against the HT-29 human colon carcinoma cell, and haouamine B does against the MS-1 mice endothelial cells. The novel molecular architecture, the kinetic aspect in the molecule, and the biological activity of haouamines atract a lot of chemists. Therefore, haouamines are challenging synthetic targets. Baran and Burns reported the synthesis of (±)-haouamine A in 2006, and the asymmetric synthesis of (+)-haouamine A in 2008. Recently Baran and co-workers accomplished the synthesis of an atropisomer of haouamine A, and indicated that the inseparable mixture oftwo interconverting isomers in solution was no due to atropisomerism. Weinreb's, Furstner's, and Ishibashi's groups have achieved the formal total synthesis. In this symposium, we report synthetic studies on haouamine B (2) by our original approach. We envisioned that the stereogenic quaternary center could be construced via an intramolecular palladium-catalyzed α-C-arylation, and the anti-Bredt doule bond introduced from a vinylogous amide by TsNHNH_2. The C2-C3 bond formation should be carried out through Suzuki-Miyaura coupling. Construction of the piperidine segment 5 could be completed by the regioselective introductoin of aryl substituents to 4-methoxypyridine (6) usin 3-methoxyphenylboronic acid (7) and 3,5-dimethoxybenzylzinc chloride (8). We also report an introducton of the stereogenic center at C17 position by Comins method., 天然有機化合物討論会実行委員会, 日本語

近年北海道で分離された牛ウイルス性下痢ウイルスの遺伝子解析
田村友和, 迫田義博, 杉田征彦, 吉野史, 岡松正敏, 喜田宏, 喜田宏, 日本獣医学会学術集会講演要旨集, 149th, 2010年

近年北海道で分離された牛ウイルス性下痢ウイルスの遺伝子と抗原性の解析
迫田義博, 田村友和, 杉田征彦, 吉野史, 岡松正敏, 喜田宏, 喜田宏, 日本ウイルス学会学術集会プログラム・抄録集, 58th, 2010年

H9N2インフルエンザウイルスはニワトリに対する病原性を獲得するか?
曽田公輔, 浅倉真吾, 岡松正敏, 迫田義博, 喜田宏, 喜田宏, 日本獣医学会学術集会講演要旨集, 149th, 2010年

A Novel Type of Hereditary Motor and Sensory Neuropathy with Proximal Dominancy in the Lower Extremities Found in a Japanese Descent
Shiroh Miura, Hiroki Shibata, Kazuhito Noda, Yoichiro Kaku, Hiroshi Kida, Akiko Iwaki, Mitsuyoshi Ayabe, Hisamichi Aizawa, Takayuki Taniwaki, ANNALS OF NEUROLOGY, 68, 4, S52, S52, 2010年
WILEY-LISS, 英語, 研究発表ペーパー・要旨（国際会議）

鳥,ブタ,そしてパンデミックインフルエンザ--季節性インフルエンザ対策の改善こそが課題 (特集 インフルエンザ)
喜田 宏, 最新医学, 65, 1, 13, 20, 2010年01月
最新医学社, 日本語

豚由来インフルエンザウイルスに対するポビドンヨード製剤の効果
伊藤 啓史, 疋田 宗生, 八代 純子, 喜田 宏, 伊藤 壽啓, 日本化学療法学会雜誌 = Japanese journal of chemotherapy, 57, 6, 508, 510, 2009年11月10日
日本語

MOUSE MODEL OF INFLUENZA VIRUS-ASSOCIATED ENCEPHALOPATHY OF CHILDHOOD
T. Tanaka, Y. Sunden, G. Tanoue, K. Ochiai, Y. Sakoda, H. Kida, T. Umemura, JOURNAL OF COMPARATIVE PATHOLOGY, 141, 4, 277, 277, 2009年11月
ELSEVIER SCI LTD, 英語, 研究発表ペーパー・要旨（国際会議）

ホモロジーモデリング法による新型H1N1インフルエンザウイルスのヘマグルチニンの抗原構造の解析
五十嵐学, 伊藤公人, 吉田玲子, 喜田宏, 喜田宏, 喜田宏, 高田礼人, 日本ウイルス学会学術集会プログラム・抄録集, 57th, 180, 2009年10月01日
日本語

インフルエンザウイルスのヘマグルチニンのMDS解析と変異予測への応用
伊藤公人, 五十嵐学, 村上悌治, 喜田宏, 喜田宏, 高田礼人, 日本ウイルス学会学術集会プログラム・抄録集, 57th, 181, 2009年10月01日
日本語

鳥,ブタ,そしてパンデミックインフルエンザ--実はすべてインフルエンザ (特集 現場の経験を活かすパンデミック対策)
喜田 宏, 月刊薬事, 51, 11, 1597, 1603, 2009年10月
じほう, 日本語

1SA2-07 鳥、ブタ、そしてパンデミックインフルエンザ(1SA2 プリオンとウィルスの感染(BSJ&ABA連携国際シンポジウム),第47回日本生物物理学会年会)
喜田 宏, 生物物理, 49, 1, S4, 2009年09月20日
日本生物物理学会, 英語

本来あるべきインフルエンザ対策とは? (新型インフルエンザの実態)
喜田 宏, メディカルバイオ, 6, 4, 75, 77, 2009年07月
オーム社, 日本語

鼎談 パンデミック前にH5N1ウイルスワクチンを接種すべきなのか,すべきではないのか
河岡 義裕, 西村 秀一, 喜田 宏, インフルエンザ, 10, 3, 191, 202, 2009年07月
メディカルレビュー社, 日本語

H5亜型ウイルスに対する鳥インフルエンザワクチンの開発 : 高力価のワクチンを開発. H7ウイルス用もスタンバイ
磯田 典和, 迫田 義博, 喜田 宏, 化学と生物, 47, 5, 302, 304, 2009年05月01日
社団法人 日本農芸化学会, 日本語

JAPHV [平成]20年度研修 特別講演 鳥インフルエンザと新型ウイルス[含 質疑応答]
喜田 宏, 獣医公衆衛生研究, 11, 2, 27, 39, 2009年03月
全国公衆衛生獣医師協議会, 日本語

最優秀賞 インフルエンザウイルスの生態解明とライブラリーの構築--高病原性鳥インフルエンザの診断と予防への応用 (畜産大賞 『研究開発部門』最優秀賞・優秀賞の紹介)
喜田 宏, 畜産技術, 0, 646, 43, 48, 2009年03月
畜産技術協会, 日本語

国内開発鳥インフルエンザワクチン
佐々木 崇, 坂元 隆一, 今村 孝, 坂口 正士, 瀧川 義康, 西條 加須江, 澤田 章, 萩原 純子, 土屋 耕太郎, 林 志鋒, 國米 則秀, 扇谷 年昭, 曽田 公輔, 磯田 典和, 迫田 義博, 喜田 宏, 鶏病研究会報, 44, 4, 170, 174, 2009年02月25日
鶏病研究会, 日本語

H9N2インフルエンザウイルスはニワトリに対する病原性を獲得するか?
曽田公輔, 浅倉真吾, 岡松正敏, 迫田義博, 喜田宏, 喜田宏, 日本ウイルス学会学術集会プログラム・抄録集, 57th, 2009年

近年流行を起こしているH3N8馬インフルエンザウイルスの遺伝子と抗原性の解析
本島昌幸, 岡松正敏, 浅倉真吾, 伊藤美加, 前田友起子, 福田奈穂, SODNOMDARJAA R., 迫田義博, 喜田宏, 喜田宏, 日本ウイルス学会北海道支部夏季シンポジウム講演抄録, 43rd, 2009年

カニクイザルを用いた高病原性鳥インフルエンザウイルスに対するワクチンの防御効果の判定
伊藤靖, 尾崎弘一, 曽田公輔, 迫田義博, 石垣宏仁, 仲山美沙子, 石田英晃, 永田智也, 三宅太一郎, 喜田宏, 喜田宏, 小笠原一誠, 日本ワクチン学会学術集会プログラム・抄録集, 13th, 2009年

今年のインフルエンザをどう乗り切るか (特集 呼吸器領域における漢方薬のエビデンス) -- (新型・季節性インフルエンザ)
岡部 信彦, 河合 直樹, 喜田 宏, 漢方医学, 33, 4, 468, 470, 2009年
臨床情報センター, 日本語

講演要旨 インフルエンザはどこから来たのか?[含 質疑応答]
喜田 宏, 読売クオータリー, 0, 10, 72, 81, 2009年
読売新聞東京本社調査研究本部, 日本語

座談会 パンデミックインフルエンザ--鳥,ブタ,そしてヒトのインフルエンザ対策の要点
喜田 宏, 西藤 岳彦, 岩崎 惠美子, Virus report, 6, 2, 12, 24, 2009年
医薬ジャーナル社, 日本語

インフルエンザウイルスの起源,進化と宿主域--新型ウイルスの出現に備えて (特集・新型インフルエンザの流行とその対策)
喜田 宏, 医薬ジャ-ナル, 44, 11, 85, 92, 2008年11月
医薬ジャ-ナル社, 日本語

H5N1不活化インフルエンザワクチンのクレード間交差防御免疫能の解析
村上晋, 岩佐彩香, 岩附(堀本)研子, 伊藤睦美, 木曽真紀, 喜田宏, 高田礼人, NIDOM Chairul A, LE QUYNH Mai, 山田晋弥, 今井博貴, 坂井(田川)優子, 河岡義裕, 堀本泰介, 日本ウイルス学会学術集会プログラム・抄録集, 56th, 224, 2008年10月01日
日本語

2008年北海道で発見された斃死オオハクチョウから分離したH5N1高病原性鳥インフルエンザウイルスの性状
岡松正敏, 迫田義博, 吉田裕美, 田中智久, 津田祥美, 磯田典和, 中山絵里, 苫米地大輔, 松野啓太, 梅村孝司, 高田礼人, 喜田宏, 日本ウイルス学会学術集会プログラム・抄録集, 56th, 263, 2008年10月01日
日本語

H5N1インフルエンザウイルスのPB2に会合する宿主細胞因子の同定とその機能解析
塩崎拓也, 岩井淳, 河岡義裕, 高田礼人, 喜田宏, 宮崎忠昭, 日本ウイルス学会学術集会プログラム・抄録集, 56th, 379, 2008年10月01日
日本語

インフルエンザウイルスHA遺伝子の塩基配列上に潜在する糖鎖付加ポテンシャルの解析
五十嵐学, 伊藤公人, 喜田宏, 喜田宏, 喜田宏, 高田礼人, 日本ウイルス学会学術集会プログラム・抄録集, 56th, 144, 2008年10月01日
日本語

2008年北海道で発見された斃死オオハクチョウから分離したH5N1高病原性鳥インフルエンザウイルスの性状
岡松正敏, 迫田義博, 吉田裕美, 田中智久, 津田祥美, 磯田典和, 中山絵里, 苫米地大輔, 松野啓太, 梅村孝司, 高田礼人, 喜田宏, 日本獣医学会学術集会講演要旨集, 146th, 193, 2008年09月05日
日本語

2007年に金沢競馬場で流行した馬インフルエンザウイルス(H3N8)の遺伝子解析(ウイルス学)
伊藤 美加, 長井 誠, 早川 裕二, 小前 博文, 村上 成人, 四ッ谷 正一, 浅倉 真吾, 迫田 義博, 喜田 宏, The journal of veterinary medical science, 70, 9, 899, 906, 2008年09月
2007年8月,石川県金沢競馬場において馬インフルエンザが発生した.感染馬の症状は主に発熱(38.2-41.0℃)および鼻汁漏出で,呼吸器症状を示した馬はわずかであった.全ての競走馬は不活化ワクチンを接種しており,ワクチンにはH3N8馬インフルエンザウイルス,アメリカ系統のA/equine/La Plata/93株,同じくヨーロッパ系統のA/equine/Avesta/93株,およびH7N7馬インフルエンザのA/equine/Newmarket/1/77株が含まれていた.鼻腔スワブ材料からH3N8馬インフルエンザが分離され,A/equine/Kanazawa/1/2007と命名した.系統樹解析では,A/equine/Kanazawa/1/2007株はアメリカ系統のフロリダ亜系統に属した.さらに,HA1サブユニットのアミノ酸解析を行ったところ,同じアメリカ系統のワクチン株であるA/equine/La Plata/93株と比較して,BおよびEの抗原決定部位に4アミノ酸の置換が認められた.また,回復期の馬16頭から採材した血清を用いて赤血球凝集抑制反応を実施したところ,A/equine/Kanazawa/1/2007株とワクチン株間に1〜3管の差が認められた.これらの結果から,流行株とワクチン株に抗原性状の差が示唆されたが,わが国の現行市販ワクチンは罹患率の低下や発症期間の短縮に貢..., 社団法人日本獣医学会, 英語

H7亜型高病原性鳥インフルエンザの迅速診断キットの開発(ウイルス学)
MANZOOR Rashid, 迫田 義博, 坂部 沙織, 望月 剛, 難波 靖治, 津田 祥美, 喜田 宏, The journal of veterinary medical science, 70, 6, 557, 562, 2008年06月
H5高病原性鳥インフルエンザウイルス(HPAIV)と同様にH7亜型のHPAIVが世界各地の家禽に甚大な被害を及ぼしている.H7HPAIVの人への感染も報告されていることから,迅速な診断がHPAIの防疫のみならず公衆衛生の観点からも鍵となる.本研究では,H7HA分子上の異なるエピトープを認識する2つのモノクローナル抗体を用いたイムノクロマトグラフフイーによる迅速診断キットを開発した.本キットは調べたすべてのH7インフルエンザウイルスを特異的に検出し,H7以外の亜型のインフルエンザウイルスおよび他のウイルスや細菌とは反応しなかった.本キットはHPAIVA/chicken/Netherland/2586/03(H7N7)に感染したニワトリのスワブおよび臓器乳剤からH7HA抗原を検出した.以上より,本迅速診断キットはH7ウイルス感染によって起こるHPAIを特異的,高感度に診断できることが判ったので,HPAI発生農場での検査に有用と考えられる., 社団法人日本獣医学会, 英語

インフルエンザウイルスの生態
喜田 宏, 臨床とウイルス, 36, 2, S19, S20, 2008年05月16日
日本語

牛ウイルス性下痢病および牛伝染性鼻気管炎に対する市販混合ワクチン接種プログラムの中和抗体応答による評価
伊藤 麻子, 迫田 義博, 亀山 健一郎, 山崎 幸夫, 臼井 章, 喜田 宏, 日本獣医師会雑誌 = Journal of the Japan Veterinary Medical Association, 61, 1, 39, 42, 2008年01月20日
牛ウイルス性下痢病（BVD）と牛伝染性鼻気管炎の予防に効果的なワクチン接種プログラムを評価するために、BVDに対して不活化抗原を含有する2つの市販混合ワクチンをそれぞれ抗体陰性牛に接種した。いずれのワクチンも初回接種後の牛にBVDウイルスおよび牛ヘルペスウイルス1に対する中和抗体がほとんど検出されなかったが、2回目の接種1ヵ月後に有意な抗体応答を認めた。その後、抗体価は時間の経過とともに漸減したが、初回接種から12ヵ月後に1回追加免疫を行うことにより抗体価が再上昇した。以上の成績より、抗体陰性牛に対する本疾病の効果的な予防には今回評価したワクチンは2回接種する必要があること、さらに年1回の追加接種が望ましいことが明らかになった。, 日本獸医師会, 日本語

日本とモンゴルで馬から分離されたH3N8インフルエンザウイルスの遺伝子と抗原性の解析
浅倉真吾, 浅倉真吾, 迫田義博, 迫田義博, 長井誠, 若本裕昌, 喜田宏, 喜田宏, 喜田宏, 日本ウイルス学会学術集会プログラム・抄録集, 56th, 2008年

日本とモンゴルで馬から分離されたH3N8インフルエンザウイルスの遺伝子および抗原性の解析
浅倉真吾, 迫田義博, 伊藤美加, 長井誠, 前田友起子, 福田奈穂, 阿部修二, BATCHULUUN D., SODNOMDARJAA R., 若本裕晶, 岡松正敏, 喜田宏, 喜田宏, 日本獣医学会学術集会講演要旨集, 146th, 2008年

H9N2インフルエンザウイルスはニワトリに対する病原性を獲得するか?
曽田公輔, 曽田公輔, 浅倉真吾, 浅倉真吾, 岡松正敏, 岡松正敏, 迫田義博, 迫田義博, 喜田宏, 喜田宏, 喜田宏, 日本獣医学会学術集会講演要旨集, 146th, 2008年

鳥インフルエンザウイルスの病原性獲得メカニズムの解析
曽田公輔, 曽田公輔, 浅倉真吾, 浅倉真吾, 岡松正敏, 岡松正敏, 迫田義博, 迫田義博, 喜田宏, 喜田宏, 喜田宏, 日本ウイルス学会学術集会プログラム・抄録集, 56th, 2008年

インフルエンザウイルスHA遺伝子の塩基配列上に潜在する糖鎖付加ポテンシャルの解析
五十嵐学, 伊藤公人, 喜田宏, 喜田宏, 喜田宏, 高田礼人, 日本ウイルス学会北海道支部夏季シンポジウム講演抄録, 42nd, 12, 2008年
日本語

アジア・アフリカ地域の鳥インフルエンザ 世界的まん延に危機感--「アジア・アフリカ地域における鳥インフルエンザの現状と国際的な取組みについてのシンポジウム」より
喜田 宏, 山田 幸雄, 藤田 陽偉, 鶏の研究, 83, 1, 108, 111, 2008年01月
木香書房, 日本語

動物インフルエンザのグローバルサーベイランスと全ての亜型ウイルスライブラリーの構築
梶原将大, 梶原将大, 迫田義博, 迫田義博, 伊藤壽啓, 高田礼人, 岸田典子, 五十嵐学, 磯田典和, 磯田典和, 曽田公輔, 曽田公輔, 喜田宏, 喜田宏, 喜田宏, 日本ウイルス学会学術集会プログラム・抄録集, 55th, 216, 2007年10月01日
日本語

インフルエンザの過去, 現在, 未来
喜田 宏, 臨床と微生物 = Clinical microbiology, 34, 5, 395, 401, 2007年09月25日
日本語

鳥インフルエンザに対するワクチンの開発
磯田 典和, 喜田 宏, ファームステージ, 7, 6, 84, 87, 2007年09月
技術情報協会, 日本語

動物インフルエンザのグローバルサーベイランスと全ての亜型ウイルスライブラリーの構築
梶原将大, 梶原将大, 迫田義博, 迫田義博, 伊藤壽啓, 高田礼人, 伊藤公人, 岸田典子, 五十嵐学, 磯田典和, 磯田典和, 曽田公輔, 曽田公輔, WEBSTER R.G, 喜田宏, 喜田宏, 喜田宏, 日本獣医学会学術集会講演要旨集, 144th, 91, 2007年08月27日
日本語

鳥インフルエンザウイルスに対する消毒薬の効果
迫田 義博, 吉見 泰, 黒川 知, 喜田 宏, 日本獣医師会雑誌 = Journal of the Japan Veterinary Medical Association, 60, 7, 519, 522, 2007年07月20日
市販消毒薬5品の鳥インフルエンザウイルス不活化効果を調べた。消毒薬はいずれもウイルスの亜型と病原性に関わらず、高希釈でウイルスの感染性を消失させた。希釈した薬液の凍結融解の繰り返しは消毒効果に影響を与えなかったが、低温下および有機物として鶏糞を添加した場合、消毒効果の減少が認められた。以上の結果から、試験に用いた消毒薬は反応温度や有機物の混入などの環境要因を十分に理解した上で適切に使用すれば鳥インフルエンザウイルスの不活化に有効であることがわかった。, 日本獸医師会, 日本語

鳥インフルエンザのワクチン開発
迫田 義博, 喜田 宏, 鶏病研究会報, 43, 1, 32, 33, 2007年05月25日
日本語

近年日本で分離された牛ウイルス性下痢ウイルスの遺伝学的および病理生物学的解析(ウイルス学)
松野 啓太, 迫田 義博, 亀山 健一郎, 玉井 久三, 伊藤 麻子, 喜田 宏, The journal of veterinary medical science, 69, 5, 515, 520, 2007年05月
2000年から2006年までに12都道県で牛から分離された475株の牛ウイルス性下痢ウイルス(BVDV)の5'末端非翻訳領域の塩基配列を基にした分子疫学解析を行った.その結果,396株が遺伝子型1に分類され,さらに1a亜型(101株),1b亜型(163株),1c亜型(128株),1j亜型(3株)およびSo CP/75-like亜型(1株)に分けられた.残り79株は遺伝子型2に分類され,すべて2a亜型であった.2a亜型BVDVには高病原性株として報告されている北米分離株と高い相同性を有する株も2株含まれていた.しかし,2a亜型BVDVのほとんどは持続感染牛から分離され,高い死亡率を伴う急性感染例は見られなかった.また,475株すべての細胞病原性とそのウイルスが分解された牛の臨床症状をまとめた結果,細胞病原性株が分離された牛でも典型的な粘膜病変を示さない例や,非細胞病原性株が分陰された持続感染牛でもまったく臨床症状を示さない例が多数認められた.以上より,国内では多様な遺伝子型のBVDVが浸潤しており,ウイルスが感染した牛が示す臨床症状も多様であることがわかった.これらの結果から,適切なワクチンによる予防と積極的なサーベイランスが本病の対策として重要であると考えられる., 社団法人日本獣医学会, 英語

人獣共通感染症の克服を目指して (特集 鳥インフルエンザを経験した京都からの発信)
喜田 宏, 養鶏の友, 541, 36, 38, 2007年03月
日本畜産振興会, 日本語

各種消毒薬の鳥インフルエンザウイルスに対する効果試験
迫田 義博, 関 令二, 喜田 宏, 家畜衛生学雑誌, 32, 2, 67, 70, 2007年03月
日本家畜衛生学会, 日本語

北海道沿岸のゼニガタアザラシ(Phoca vitulina stejnegeri)のインフルエンザAウイルス感染に関する血清学的調査(ウイルス学)
藤井 啓, 角本 千治, 小林 万里, 齋藤 幸子, 刈屋 達也, 渡邊 有希子, 迫田 義博, 喜田 宏, 鈴木 正嗣, The journal of veterinary medical science, 69, 3, 259, 263, 2007年03月
ゼニガタアザラシの保護管理に役立てることを目的として,北海道の太平洋沿岸に生息する野生ゼニガタアザラシの,インフルエンザAウイルス感染に関する血清学的調査を行った.1998年から2005年にかけて,納沙布で231個体,厚岸で16個体,襟裳で75個体のサンプル血清が採取された.ELISAでは,納沙布のサンプルからのみ,インフルエンザAウイルスに対する抗体が検出された.抗体保有率は98年が11%(1/9),03年が3%(2/66),04年が12%(7/59),05年が6%(5/77)であり,散発的な感染の存在が示された.いずれの年にも,幼獣からの抗体の検出があったため,少なくとも90年代の終盤から近年まで,感染が繰り返されていると考えられた.HI検査によると,ELISA陽性であった15サンプルのうち,10サンプルからH3亜型に対する抗体が,2サンプルからH6亜型に対する抗体が検出された.H6亜型に対して陽性であった2サンプルはH3亜型に対しても陽性であり,この2個体はH3とH6亜型の両方に感染したと考えられる.H6亜型に対する抗体がアザラシ類で確認されたのは,本報告が初めてである.H6亜型はこれまで鳥類からのみ分離されているが,遺伝学的研究では哺乳類に感染する可能性が示唆されている.今後,ウイルス検出による確定診断を試みたい., 社団法人日本獣医学会, 英語

インフルエンザウイルス亜型間交差中和活性を有するモノクローナル抗体の解析
吉田玲子, 五十嵐学, 喜田宏, 喜田宏, 喜田宏, 高田礼人, 日本ウイルス学会学術集会プログラム・抄録集, 54th, 347, 2006年11月01日
日本語

グローバルサーベイランスの重要性 (特集 インフルエンザをめぐる諸問題)
尾崎 弘一, 喜田 宏, アレルギ-・免疫, 13, 11, 1530, 1535, 2006年11月
医薬ジャ-ナル社, 日本語

Efficacy of intracerebral vaccination against pseudorabies virus in mice
Jae-Ho Shin, Yoshihiro Sakoda, Jae Hoon Kim, Tomohisa Tanaka, Hiroshi Kida, Takashi Kimura, Kenji Ochiai, Takashi Umemura, JOURNAL OF NEUROIMMUNOLOGY, 178, 114, 114, 2006年09月
ELSEVIER SCIENCE BV, 英語, 研究発表ペーパー・要旨（国際会議）

2003年から2005年にベトナムで分離されたA型H5N1インフルエンザウイルスの病原性解析(短報)(ウイルス学)
村本 裕紀子, LE Thi Quynh Mai, PHUONG Lien Song, NGUYEN Tung, NGUYEN Thu Ha, 坂井(田川) 優子, 堀本 泰介, 喜田 宏, 河岡 義裕, The journal of veterinary medical science, 68, 7, 735, 737, 2006年07月
2003年以降,アジアで高病原性H5N1鳥インフルエンザが流行し,特にベトナムではヒトでの感染例が多く報告されている.これらウイルスの哺乳類に対する病原性を調べるため,ベトナムのヒト,ニワトリ,アヒルから分離されたウイルスをマウスに実験感染させた.その結果,ヒト由来株は2株とも,家禽由来株は5株中1株がマウスを殺す強毒株であった.ベトナムでは家禽の間で哺乳類に致死的な感染をおこすウイルスが流行していることがわかった., 社団法人日本獣医学会, 英語

2004年および2005年にベトナムの家禽および水禽から分離されたA型H5N1鳥インフルエンザウイルスのヘマグルチニンならびにノイラミニダーゼ遺伝子の系統解析(短報)(ウイルス学)
村本 裕紀子, LE Thi Quynh Mai, PHUONG Lien Song, NGUYEN Tung, NGUYEN Thu Ha, 坂井(田川) 優子, 岩附(堀本) 研子, 堀本 泰介, 喜田 宏, 河岡 義裕, The journal of veterinary medical science, 68, 5, 527, 531, 2006年05月
2003年以降,アジアで高病原性H5N1鳥インフルエンザウイルスが流行している.最大の流行地であるベトナムで流行しているウイルスの性状を調べるため,2004年および2005年にアヒルとニワトリから分離されたH5N1鳥インフルエンザウイルスのヘマグルチニンならびにノイラミニダーゼ遺伝子の系統解析を行った.その結果,2004年および2005年の間に,異なる系統のH5N1ウイルスが度々ベトナムに侵入していることが示唆された., 社団法人日本獣医学会, 英語

鳥インフルエンザウイルスについて (特集 高病原性鳥インフルエンザについて)
喜田 宏, 畜産技術, 0, 611, 6, 11, 2006年04月
畜産技術協会, 日本語

人獣共通感染症としての高病原性鳥インフルエンザとワクチン (特集 インフルエンザウイルス感染症と関連するワクチンの臨床) -- (インフルエンザウイルス感染症の特殊な病態と合併症)
喜田 宏, 日本医師会雑誌, 134, 10, 1940, 1944, 2006年01月
日本医師会, 日本語

A型およびB型インフルエンザウイルスを検出する迅速診断キットの改良(ウイルス学)
白 貴蓉, 迫田 義博, ムウイネ アーロン, 藤井 信之, 皆川 英孝, 喜田 宏, The journal of veterinary medical science, 68, 1, 35, 40, 2006年01月
A型およびB型インフルエンザウイルスを特異的に検出する迅速診断キットの感度を向上させるため, A型ウイルスNP蛋白の59-130番アミノ酸領域およびB型ウイルスNP蛋白の72-191番アミノ酸領域を認識するモノクローン抗体を用いたエスプライン^[○!R]インフルエンザA&B-Nを開発した.本キットの検出限界は10^<2.0>-10^<2.7> pfu/テストであり, 従来キットに比べ, 4〜1,000倍感度が改善された.また本キットは近年アジア地域でヒトや家禽から分離された高病原性H5N1ウイルスを含むすべてのHA亜型のA型ウイルスならびに調べた15株すべてのB型ウイルス株と反応した.以上の成績から, 今回開発したエスプライン^[○!R]インフルエンザA&B-NはA型およびB型ウイルスを迅速かつ高感度に検出することがわかった., 社団法人日本獣医学会, 英語

インフルエンザウイルスの生態
喜田 宏, 日本豚病研究会報 = Proceedings of the Japanese Pig Veterinary Society, 47, 16, 17, 2005年08月01日
日本豚病研究会, 日本語

鼎談 ウイルスとその宿主をとりまく環境
河岡 義裕, 倉根 一郎, 喜田 宏, Virus report, 2, 1, 8, 13, 2005年06月
医薬ジャーナル社, 日本語

鳥インフルエンザウイルスの生態(高病原性鳥インフルエンザとクロバエ類の関わり)
喜田 宏, 衞生動物, 56, 0, 30, 30, 2005年06月01日
日本衛生動物学会, 日本語

座談会 Avian influenza (鳥インフルエンザ特集号)
Webster R. G, 永井 美之, 喜田 宏, インフルエンザ, 6, 2, 113, 124, 2005年04月
メディカルレビュー社, 日本語

エボラウイルス・ヌクレオカプシド様構造物の電子顕微鏡トモグラフィー法による立体再構築(短報)(ウイルス学)
野田 岳志, 青山 一弘, 相良 洋, 喜田 宏, 河岡 義裕, The journal of veterinary medical science, 67, 3, 325, 328, 2005年03月
電子顕微鏡トモグラフィー法は, 非対称性および対称性高分子複合体の三次元再構築を可能にした新しい技術である.我々は本法を用いて, エボラウイルス・ヌクレオカプシド様構造物の立体再構築を行い, 形態学的解析を行った.その結果, チューブ状のヌクレオカプシド様構造物は, 他の非分節化マイナス鎖RNAウイルスと同様に, 左巻き螺旋構造であることが明らかになった., 社団法人日本獣医学会, 英語

ポビドンヨード製剤の抗鳥インフルエンザウイルス活性
伊藤 啓史, 疋田 宗生, 大塚 昭, 八代 純子, 喜田 宏, 大槻 公一, 伊藤 壽啓, 日本化学療法学会雜誌 = Japanese journal of chemotherapy, 53, 1, 20, 22, 2005年01月25日
日本語

2P012 フェニル酢酸の異化に関わるPaaタンパク質群の結晶学的研究(蛋白質 A) 構造))
木田 宗志, 飯野 均, 中川 紀子, 久野 玉雄, 藤橋 雅宏, 喜田 昭子, 三木 邦夫, 生物物理2005年, 45, supplement, S122, S122, 2005年
Biophysical Society of Japan, 日本語

特集 2005年新年号座談会:新興・再興感染症の現状と防疫をめぐって
喜田 宏, 栗本 まさ子, 高崎 智彦, 畜産技術, 0, 596, 2, 19, 2005年01月
畜産技術協会, 日本語

基礎から臨床へ 動物からヒトへ--人獣共通感染症を克服するために
喜田 宏, 診断と治療, 92, 12, 2289, 2295, 2004年12月
診断と治療社, 日本語

新型インフルエンザと鳥インフルエンザ (特集:エマージング感染症の時代)
喜田 宏, 現代化学, 0, 404, 27, 31, 2004年11月
東京化学同人, 日本語

パネルディスカッション 平成16年度北日本養鶏研究大会より(3)高病原性鳥インフルエンザ--ワクチン接種の効果と論点について
所 源亮, 喜田 宏, 境 政人, 鶏卵肉情報, 34, 20, 34, 37, 2004年10月25日
鶏卵肉情報センタ-, 日本語

パネルディスカッション 平成16年度北日本養鶏研究大会より(2)高病原性鳥インフルエンザ--ワクチン接種の効果と論点について
所 源亮, 喜田 宏, 境 政人, 鶏卵肉情報, 34, 19, 18, 23, 2004年10月10日
鶏卵肉情報センタ-, 日本語

鳥インフルエンザとは何か (特集 人と動物の共通感染症(1)鳥インフルエンザ)
喜田 宏, 公衆衛生, 68, 10, 760, 763, 2004年10月
医学書院, 日本語

パネルディスカッション 平成16年度北日本養鶏研究大会より(1)高病原性鳥インフルエンザ--ワクチン接種の効果と論点について
所 源亮, 喜田 宏, 境 政人, 鶏卵肉情報, 34, 18, 38, 44, 2004年09月25日
鶏卵肉情報センタ-, 日本語

鳥インフルエンザと新型インフルエンザ
喜田 宏, 日本臨床免疫学会会誌 = Japanese journal of clinical immunology, 27, 4, 242, 242, 2004年08月31日
日本臨床免疫学会, 日本語

人獣共通感染症の克服に向けて
喜田 宏, 日本獣医師会雑誌 = Journal of the Japan Veterinary Medical Association, 57, 8, 471, 476, 2004年08月20日
日本獣医師会, 日本語

座談会「高病原性鳥インフルエンザをめぐって」
永井 美之, 大槻 公一, 岡部 信彦, 河岡 義裕, 喜田 宏, 倉田 毅, 佐多 徹太郎, 田代 眞人, 山口 成夫, 吉倉 廣, ウイルス, 54, 1, 123, 141, 2004年06月30日
日本ウィルス学会, 日本語

鳥インフルエンザウイルス
喜田 宏, ウイルス, 54, 1, 93, 96, 2004年06月30日
2003年から2004年にかけて日本を含むアジア9カ国で発生した一連の高病原性鳥インフルエンザは, 感染症に国境がないこと, 日本の鶏群にも本症発生のリスクがあることに加えてウイルスの侵入をいち早く検出するために年間を通した家禽のモニタリングが必須であることを改めて諭す教訓となった. アジアでは今なお高病原性鳥H5N1インフルエンザウイルスによる家禽の被害が断続している. さらにタイとベトナムではそれぞれ12名および22名のヒトがH5N1ウイルスに感染し, 8名および15名が死亡した. 今後, 再び高病原性鳥インフルエンザが日本に飛び火する可能性を否定できない状況にある. 家禽における鳥インフルエンザウイルスの感染を早期に摘発, 淘汰することによって家禽の被害を最小限にくい止めるとともに, ヒトの健康と食の安全を守ることが鳥インフルエンザ対策の基本である., 日本ウィルス学会, 日本語

鳥インフルエンザウイルスと新型インフルエンザウイルス
喜田 宏, 臨床とウイルス, 32, 2, S18, 2004年05月31日
日本語

鳥インフルエンザワクチン接種是非論争 ワクチン接種すべきでない
喜田 宏, 週刊農林, 0, 1881, 8, 8, 2004年03月15日
農林出版社, 日本語

Subcutaneous hematoma due to frequent insulin injections in a single site
Toshio Kahara, Shigeru Kawara, Akiko Shimizu, Azusa Hisada, Yutaka Noto, Hiroshi Kida, Internal Medicine, 43, 2, 148, 149, 2004年02月
英語, 速報，短報，研究ノート等（学術雑誌）

2P011 高度好熱菌由来4-oxalocrotonate tautomeraseの結晶構造(蛋白質 A) 構造)
木田 宗志, 喜田 昭子, 三木 邦夫, 生物物理2004年, 44, supplement, S112, S112, 2004年
Biophysical Society of Japan, 日本語

Development of malignancy after treatment of idiopathic membranous nephropathy [7] (multiple letters)
Ming-Y.U. Lai, W.U.-Chang Yang, Chih-Ching Lin, Keiichi Yoshimoto, Hitoshi Yokoyama, Takashi Wada, Kengo Furuichi, Satoshi Goshima, Hiroshi Kida, Kidney International, 66, 1, 464, 465, 2004年
Blackwell Publishing Inc., 英語, 速報，短報，研究ノート等（学術雑誌）

Avian influenza viruses as the origin of pandemic strains
Hiroshi Kida, JOURNAL OF NEUROVIROLOGY, 14, 14, 14, 2004年
TAYLOR & FRANCIS INC, 英語, 研究発表ペーパー・要旨（国際会議）

インフルエンザパンデミックに対する危機管理体制と国際対応に関する研究 インフルエンザ株サーベイランスと新型ワクチン開発,国際協力の検討
小田切孝人, 西藤岳彦, 板村繁之, 小淵正次, 今井正樹, 二宮愛, 山下和予, 谷口清洲, 喜田宏, インフルエンザパンデミックに対する危機管理体制と国際対応に関する研究 平成15年度 総括・分担研究報告書, 34, 37, 2004年
日本語

Host range of influenza viruses and the development of new anti influenza agents
Y Suzuki, IPJ Hidari, D Miyamoto, T Suzuki, CT Guo, T Ito, H Kida, Y Kawaoka, JOURNAL OF CLINICAL VIROLOGY, 28, S104, S105, 2003年12月
ELSEVIER SCIENCE BV, 英語, 研究発表ペーパー・要旨（国際会議）

鳥インフルエンザウイルスの感染性に及ぼす消毒薬の効果について
伊藤 壽啓, 喜田 宏, 大槻 公一, 家畜衛生学雑誌, 29, 2, 123, 126, 2003年12月
日本家畜衛生学会, 日本語

新型インフルエンザウイルス襲来 (世界を脅かす感染症とどう闘うか) -- (第1章 広がる脅威)
野田 岳志, 喜田 宏, 河岡 義裕, 別冊日経サイエンス, 143, 10, 13, 2003年11月
日経サイエンス, 日本語

マウスおよび鶏におけるインフルエンザウイルス性脳炎発生への解熱剤投与の影響(病理学)
寸田 祐嗣, 朴 天鎬, 松田 一哉, 穴川 明子, 木村 享史, 落合 謙爾, 喜田 宏, 梅村 孝司, The journal of veterinary medical science, 65, 11, 1185, 1188, 2003年11月
インフルエンザウイルス性脳炎に対する解熱剤投与の影響を検討するためにマウスと鶏のインフルエンザ脳炎モデル動物に対して,アセチルサリチル酸(アスピリン)またはジクロフェナクナトリウム(ボルタレン)を多量投与してその影響を観察した.マウスでは解熱剤投与の影響は組織学的または免疫組織学的には表れなかった.一方,鶏では解熱剤を投与した鶏だけに血管周囲性細胞浸潤とグリオーシスが認められ,さらにウイルス抗原はボルタレン投与鶏のみのグリア細胞または血管内皮細胞に検出された.本実験結果より解熱剤投与はマウスにおけるインフルエンザウイルスの経神経感染には影響しないが,鶏での血行性感染を増強することが示唆された., 社団法人日本獣医学会, 英語

第50回日本ウイルス学会学術集会・総会開催記
喜田 宏, ウイルス, 53, 1, 93, 94, 2003年06月01日
日本語

新型インフルエンザウイルス対策
喜田 宏, ウイルス, 53, 1, 71, 74, 2003年06月01日
日本ウィルス学会, 日本語

牛ウイルス性下痢ウイルス非構造蛋白NS3に対するモノクローン抗体によるペスチウイルス属共通エピトープの検出
玉井久三, 迫田義博, 青木博史, 中村成幸, 喜田宏, 日本獣医学会学術集会講演要旨集, 135th, 131, 131, 2003年03月25日
(公社)日本獣医学会, 日本語

ヘルペスウイルスgBのα‐helix領域ペプチドによるウイルスの増殖阻害
岡崎克則, 喜田宏, 日本獣医学会学術集会講演要旨集, 135th, 121, 2003年03月25日
日本語

犬ヘルペスウイルス前初期遺伝子の転写動態の解析(ウイルス学)
三好 正浩, 岡崎 克則, 滝口 満喜, 喜田 宏, 橋本 晃, The journal of veterinary medical science, 64, 7, 627, 631, 2002年07月
犬ヘルペスウイルス(CHV)の前初期(IE)遺伝子は,単純ヘルペスウイルス1の感染細胞内蛋白質4(ICP4)遺伝子に相同な遺伝子で,約4.9kbのmRNAとして転写される.今回我々は,CHV IE遺伝子が感染の初期および後期には約4.8kbのmRNAとして転写される知見を得た.この4.8kbのmRNAは,後期にはIE期プロモーターからの転写が抑制されるIE期転写物のイントロン中に存在するTATAボックスの下流から転写されていた.今回の成績から,CHV IE遺伝子はCICP4遺伝子と改称することが適当であると考えられる., 社団法人日本獣医学会, 英語

人獣共通感染症としてのインフルエンザ : 新型ウイルスの出現メカニズムと対策
喜田 宏, 獣医疫学雑誌 = The journal of veterinary epidemiology, 6, 1, 29, 33, 2002年06月20日
1968年に出現した新型インフルエンザウイルスA/ホンコン/68 (H3N2) 株はカモがシベリアの営巣湖沼から家禽に持ち込んだウイルスとそれまでヒトに流行していたH2N2ウイルスが中国南部でブタの呼吸器に共感染して生じた遺伝子再集合体である。1957年の新型H2N2ウイルスも同様の経路で出現したものと推定される。1918年にスペインインフルエンザを引き起こしたH1N1ウイルスは北米大陸の系統の鳥類インフルエンザウイルスを起源とする。
1997年に香港で18人に感染して6人を死亡させたH5N1ウイルスもまた, シベリアからカモが家禽に持ち込んだウイルスがニワトリに受け継がれる問に病原性を獲得したものである。1983年に米国ペンシルバニア州でH5N2ウイルスによる家禽ペストが発生して以来, 世界各地で家禽にインフルエンザが発生していることが判ってきた。水禽の弱毒ウイルスがニワトリに感染して伝播すると, ニワトリに対して病原性を獲得することがあり, これが家禽ペストを引き起こす。香港H5N1ウイルス事件はまた, 生鳥の小売りマーケットがインフルエンザウイルスの遺伝子プールであることを明らかにした。
水禽, その営巣湖沼水, 家禽と家畜の疫学調査を強化し, 新型ウイルスの出現とその亜型を予測すると共に, 調査で分離されるウイルスの中から的確なワクチン株および診断抗原を選出して, 供給する計画がWHO, 日米医学協力研究会, 厚生労働省ならびに文部科学省に採択され, 動物インフルエンザのグローバルサーベイランスを推進している。, 獣医疫学会, 日本語

Electron-dense deposition patterns and the outcomes of idiopathic membranous nephropathy in Japanese
H. Yokoyama, Keiichi Yoshimoto, Takashi Wada, Kengo Furuichi, Satoshi Goshima, Hiroshi Kida, Ken-Ichi Kobayashi, Medical Electron Microscopy, 35, 2, 81, 86, 2002年06月
A considerable diversity in prognosis is seen with membranous nephropathy (MN). In terms of pathological findings, the presence of tubulointerstitial lesions was emphasized as a poor prognostic factor. However, the glomerular factors affecting the long-term outcome of idiopathic human MN have remained unclear. We reviewed the initial clinicopathological factors affecting the primary and secondary outcomes in 105 patients with primary MN, as well as reviewing previous reports. Based on electron microscopic (EM) findings, we could divide patients into two subtypes and one subgroup
i.e., homogeneous type with a synchronous phase of electron-dense deposits, with large dense deposits (deep subgroup) and heterogeneous type with various phases of dense deposits. The homogeneous type showed no endstage renal failure, and had earlier remission as compared with the heterogeneous type. For the secondary outcome, heterogeneous type and deep sub-group were also independent risk factors. However, there was no significant difference in the final primary or secondary outcome for any treatment subgroups. These results indicated that our category of EM findings was a beneficial marker of the primary and secondary outcomes in MN
the homogeneous type of MN with synchronous phase of electron-dense deposits (except for the "deep" subgroup) had a good outcome., 英語, 書評論文,書評,文献紹介等

インフルエンザウイルスのリバースジェネティクス--ウイルス改造現場からのレポート (特集 感染症発症の分子機構 宿主と病原体の分子の攻防)
八田 正人, 渡辺 登喜子, 喜田 宏, モレキュラ-メディシン, 39, 2, 188, 195, 2002年02月
中山書店, 日本語

基礎 自然界におけるゲノムプールと新型インフルエンザウイルス (特集 微生物ゲノム情報と医学--基礎と臨床)
喜田 宏, 現代医療, 34, 5, 1041, 1046, 2002年
現代医療社, 日本語

鼎談 新型インフルエンザウイルスの出現とインフルエンザ脳炎・脳症
喜田 宏, 富樫 武弘, 河岡 義裕, インフルエンザ, 2, 3, 191, 206, 2001年07月
メディカルレビュー社, 日本語

基礎 新型インフルエンザウイルスの出現に備える
喜田 宏, インフルエンザ, 2, 2, 127, 135, 2001年04月
メディカルレビュー社, 日本語

「新型」ウイルス出現のメカニズムとその流行予防対策 (特集 インフルエンザ) -- (21世紀のインフルエンザ)
喜田 宏, 小児科診療, 63, 12, 2138, 2142, 2000年12月
診断と治療社, 日本語

インフルエンザウイルスの遺伝子操作系の確率と応用 - 誰でもできる超簡単法 -
渡辺 登喜子, 喜田 宏, 河岡 義裕, ウイルス, 50, 1, 1, 9, 2000年06月01日
A型インフルエンザウイルスは, 8本のマイナス鎖のRNA分節を遺伝子として持つウイルスである。最近, 我々は細胞内に存在するRNAポリメラーゼを利用することにより, 効率よくインフルエンザウイルス様粒子 (virus-like particles: VLP) を作出する系を確立し, さらに, その遺伝子が全てcDNAに由来するインフルエンザウイルスを生成する遺伝子操作技術を開発した。この技術を用いて, 細胞に1度だけ感染しウイルス蛋白質は発現するが, 新たな感染性粒子は産生しない"半生"インフルエンザワクチンを設計した。その効果をマウスを用いて検定したところ,"半生"ワクチンは顕著な感染防御効果を示した。このように, 新しい遺伝子操作技術はインフルエンザウイルスを自由に設計することを可能とし, 基礎研究のみならずワクチンやベクター開発に広く貢献することが期待される。, 日本ウイルス学会, 日本語

イヌヘルペスウイルス(CHV)感染新生子犬の脳病変におけるウイルス抗原及びウイルスゲノムの分布
西野亜紀, 滝口満喜, 橋本晃, 岡崎克則, 喜田宏, 日本獣医学会学術集会講演要旨集, 129th, 47, 2000年03月01日
日本語

ウシヘルペスウイルス1のリセプター特異性
岡崎克則, 高田礼人, 喜田宏, 日本獣医学会学術集会講演要旨集, 129th, 86, 2000年03月01日
日本語

ウマヘルペスウイルス1型脳脊髄炎のマウスを用いた実験的再現
佐藤栄一, 梅村孝司, 落合謙爾, 向谷竜介, 岡崎克則, 喜田宏, 和田隆一, 日本獣医学会学術集会講演要旨集, 129th, 41, 2000年03月01日
日本語

インフルエンザウイルスの分子生物学2 宿主域とレセプター特異性
喜田宏, 治療学, 34, 33, 37, 2000年

インフルエンザは人獣共通伝染病-新型ウイルス出現のメカニズムと流行予防
喜田宏, 綜合臨床, 49, 252, 255, 2000年

新型インフルエンザウイルス出現のしくみと対策 (免疫と感染・癌)
喜田 宏, 免疫, 2000, 0, 228, 235, 2000年
中山書店, 日本語

新型インフルエンザウイルスはどこからくるのか:出現のメカニズムと流行の予防 (特集 インフルエンザ 最強のウイルスの謎に迫る)
喜田 宏, 細胞工学, 19, 1, 27, 32, 2000年01月
秀潤社, 日本語

動物とヒトのインフルエンザ--新型ウイルス出現と抗原変異
喜田 宏, 動生協会々報, 32, 4, 1, 10, 1999年10月
動物用生物学的製剤協会, 日本語

イヌヘルペスウイルスの神経節神経細胞およびリンパ節リンパ球における潜伏感染
三好 正浩, 石井 由紀, 滝口 満喜, 高田 礼人, 安田 隼, 橋本 晃, 岡崎 克則, 喜田 宏, The journal of veterinary medical science, 61, 4, 375, 379, 1999年04月
イヌヘルペスウイルス(CHV)の潜伏感染部位を明らかにするため, CHVを実験感染後ウイルス排泄が認められなくなったイヌの主な組織を調べた. これらの組織中に感染性ウイルスおよびウイルス抗原は検出されなかった. Nested PCRによる検索の結果, 三叉神経節および咽頭後リンパ節において高率にウイルスゲノムが検出された. それらの組織をin situ ハイブリダイゼーションに供したところ, ウイルスDNAは, それぞれ神経細胞およびリンパ球の核に検出された., 社団法人日本獣医学会, 英語

新型インフルエンザウイルスの出現メカニズムとその対策 (主題 かぜ症候群)
喜田 宏, 小児科診療, 62, 3, 340, 345, 1999年03月
診断と治療社, 日本語

新型インフルエンザの発生メカニズムと対策 (特集 感染症研究)
喜田 宏, 学術月報, 52, 2, 182, 187, 1999年02月
日本学術振興会, 日本語

東シベリア先住民とイヌイットから検出されたコーカソイド系JCウイルス
杉本智恵, DEMENEV V, 関野吉晴, 本庄健男, 小嶋一男, 北村唯一, 加藤温, 喜田宏, 余郷嘉明, 日本分子生物学会年会プログラム・講演要旨集, 22nd, 1999年

インフルエンザの動物種間伝播 : 新型ウイルスの出現に果たすカモ, 家禽とブタの役割
喜田宏, 化学療法の領域, 15, 29, 34, 1999年

インフルエンザの動物間伝搬 : 新型ウイルス出現に果たすカモ、家禽とブタの役割
喜田宏, 化学療法の領域, 15, 1365, 1370, 1999年

遺伝子組換えワクチンの安全性試験法の開発 (厚生省S)
田代真人, 板村繁之, 西村秀一, 西藤岳彦, 榎並正芳, 喜田宏, 新型インフルエンザワクチンの開発・製造・品質管理に関する研究研究報告書 平成10年度, 4, 14, 1999年
日本語

リバースジェネテイクスを利用した新型インフルエンザウイルス(H5N1)に対するワクチン製造候補株の作製
板村 繁之, 西村 秀一, 重茂 克彦, 根路銘 国昭, 浮田 敏幸, 神田 忠仁, 相崎 英樹, 松浦 善治, 宮村 達男, 竹内 薫, 棚林 清, 西藤 岳彦, 田代 眞人, 山崎 修道, 榎並 正芳, 榎並 和枝, 岡崎 克則, 喜田 宏, 日本分子生物学会年会プログラム・講演要旨集, 21, 0, 340, 340, 1998年12月01日
日本語

JCウイルスからみた東シベリア先住民の起源
杉本 智恵, DEMENEV Vladimir, 本庄 健男, 小嶋 一男, 喜田 宏, 小林 伸好, 余郷 嘉明, 日本分子生物学会年会プログラム・講演要旨集, 21, 0, 345, 345, 1998年12月01日
日本語

新型インフルエンザウイルスの出現予測と流行防止
喜田 宏, 科学, 68, 9, 691, 699, 1998年09月
岩波書店, 日本語

中国のブタにおける鳥インフルエンザウイルスのヘマグルチニンに対する抗体の検出
二宮愛, 安田二朗, 高田礼人, 岡崎克則, SHORTRIDGE K F, 喜田宏, 日本獣医学会学術集会講演要旨集, 126th, 142, 1998年08月
日本語

ウシヘルペスウイルス1gB糖蛋白の機能
岡崎克則, 藤井早苗, 喜田宏, 日本獣医学会学術集会講演要旨集, 126th, 110, 1998年08月
日本語

イヌヘルペスウイルスの潜状感染部位の同定
三好正浩, 石井由紀, 滝口満喜, 安田準, 橋本晃, 高田礼人, 岡崎克則, 喜田宏, 日本獣医学会学術集会講演要旨集, 125th, 140, 1998年03月
日本語

人獣共通伝染病・インフルエンザの疫学
喜田 宏, 家畜診療, 414, 3, 14, 1997年12月
全国農業共済協会, 日本語

イヌヘルペスウイルス前初期遺伝子の構造
三好正浩, 滝口満喜, 安田準, 橋本晃, 岡崎克則, 喜田宏, 日本獣医学会学術集会講演要旨集, 124th, 150, 1997年09月
日本語

ウシヘルペスウイルス1の細胞内侵入機構の解析
永野登喜子, 岡崎克則, 喜田宏, 日本獣医学会学術集会講演要旨集, 124th, 149, 1997年09月
日本語

ウシヘルペスウイルス1の細胞侵入過程におけるヘパリン様物質の関与
岡崎克則, 大塚治城, 喜田宏, 日本獣医学会学術集会講演要旨集, 124th, 149, 1997年09月
日本語

エボラウイルス表面糖蛋白の機能解析 プラスミドによる発現とG蛋白欠失VSVの利用
高田礼人, SANCHEZ A, WHITT M A, 喜田宏, 岡崎克則, 河岡義裕, 日本獣医学会学術集会講演要旨集, 124th, 147, 1997年09月
日本語

Sialyl-linkage mediated selection for the appearance of host cell variants of influenza A viruses
Y Suzuki, T Ito, H Masuda, A Takada, A Kawamoto, K Otsuki, D Miyamoto, T Suzuki, H Kida, Y Kawaoka, FASEB JOURNAL, 11, 9, A1443, A1443, 1997年07月
FEDERATION AMER SOC EXP BIOL, 英語, 研究発表ペーパー・要旨（国際会議）

Laboratory of Microbiology
KIDA Hiroshi, OKAZAKI Katsunori, TAKADA Ayato, Japanese journal of veterinary research, 45, 1, 32, 33, 1997年05月30日
北海道大学, 英語

バキュロウイルスをベクターとしたほ乳動物細胞における組換え蛋白の発現
青木博史, 迫田義博, 土屋佳紀, 勝二郁夫, 松浦善治, 高田礼人, 喜田宏, 杉山公宏, 福所秋雄, 日本獣医学会学術集会講演要旨集, 123rd, 153, 1997年03月
日本語

抗NS1抗体の検出によるインフルエンザの血清診断法
尾崎弘一, 岡崎克則, 伊藤寿啓, 今川浩, 杉田繁夫, 喜田宏, 日本獣医学会学術集会講演要旨集, 123rd, 150, 1997年03月
日本語

シベリアの水禽営巣地におけるインフルエンザウイルスの分布
岡崎克則, 伊藤寿啓, 高桑弘樹, 今井正樹, 八田正人, DEMENEV V A, 喜田宏, 日本獣医学会学術集会講演要旨集, 123rd, 151, 1997年03月
日本語

インフルエンザウイルスの生態--新型ウイルスの出現に備えて
喜田 宏, 蛋白質核酸酵素, 42, 2, 145, 153, 1997年02月
共立出版, 日本語

動物界におけるインフルエンザウイルスの生態と新型ウイルス出現のメカニズム
喜田宏, 日本臨床, 55, 2521, 2526, 1997年

N-acetyl neuraminic acid-2,6-galactoselinkage is abundantin amnion but not in allantois of chicken eggs: Implication on efficient isolation of human influenza viruses in the former compartment and selection of egg variants　　　　　　　　　　　　　　　
Ito, T, Suzuki, T, Takada, A, Kawamoto, A, Otsuki, K, Masuda, H, Yamada, M, Suzuki, Y, Kida, H, Kawaoka, Y, Journal of Virology, 71, 3357, 3362, 1997年

Viral diseases of livestock in Zambia
Aaron S. Mweene, Girja S. Pandey, Peter Sinyangwe, Andrew Nambota, Kenny Samui, Hiroshi Kida, Japanese Journal of Veterinary Research, 44, 2, 89, 105, 1996年08月
This review is to provide information on viral diseases of livestock in Zambia. The distribution of the diseases as well as the control measures and limited research that has been done, are described. Foot and mouth disease (FMD) causes serious economic losses in the cattle industry. So far five serotypes (SAT1, SAT2, SAT3, O and A) of FMD virus have been isolated in Zambia. Other notifiable viral diseases are rabies, Rift Valley fever, lumpy skin disease, African horse sickness, bluetongue, African swine fever, Newcastle disease, Marek's disease, fowlpox and infectious bursal disease. Based on the reports of clinical and/or serological diagnoses, these are widespread in the country, although their precise incidence rates are not known. With the establishment of a veterinary school equipped with modern diagnostic facilities and the increasing number of qualified veterinary personnel, this review would stimulate surveillance study on the viral diseases for the ultimate goal of achieving effective disease control measures., 英語, 書評論文,書評,文献紹介等

インフルエンザウイルスRNAポリメラーゼPB2の構造と機能
八田正人, 升永憲治, 伊藤寿啓, 岡崎克則, 豊田哲也, 石浜明, 喜田宏, 日本獣医学会学術集会講演要旨集, 122nd, 89, 1996年08月
日本語

抗ヘマグルチニン抗体によるインフルエンザウイルスの感染性中和の機序
今井正樹, 伊藤寿啓, 岡崎克則, 喜田宏, 日本獣医学会学術集会講演要旨集, 122nd, 88, 1996年08月
日本語

牛伝染性鼻気管炎感染防御ワクチンに関する研究
矢野淳, アーロンムウイネ, 今井正樹, 尾崎弘一, 永野登喜子, 二宮愛, 伊藤寿啓, 岡崎克則, 喜田宏, 日本獣医学会学術集会講演要旨集, 122nd, 191, 1996年08月
日本語

インフルエンザA型ウイルスのシアル酸分子種に対する結合特異性と宿主粘膜表皮細胞のシアル酸組成
堀池 剛, 鈴木 隆, 増田 裕幸, 山崎 泰広, 山田 未佳, 宮本 大誠, 木曽 真, 長谷川 明, 伊藤 壽啓, 喜田 宏, 河岡 義裕, 鈴木 康夫, 日本分子生物学会年会プログラム・講演要旨集, 19, 0, 90, 90, 1996年08月
日本語

インフルエンザウイルスの宿主変異と受容体シアロ糖鎖認識
鈴木 康夫, 山田 未佳, 増田 裕幸, 堀池 剛, 鈴木 隆, 宮本 大誠, 木曾 真, 長谷川 明, 伊藤 壽啓, 喜田 宏, 河岡 義弘, WEBSTER R. g, 日本分子生物学会年会プログラム・講演要旨集, 19, 0, 391, 391, 1996年08月01日
日本語

1993年に豚から分離されたA型インフルエンザウイルス(H3N2)の性状
勝田 賢, 白幡 敏一, 喜田 宏, 後藤 仁, The journal of veterinary medical science, 57, 6, 1023, 1027, 1995年12月
1993年に豚から分離された3株の香港型インフルエンザウイルスのヘムアグルチニン(HA)について, その抗原性ならびに遺伝子解析を行った. 香港型ウイルスに対する抗血清ならびにHAに対するモノクローナル抗体を用いたHI試験の成績より, 分離ウイルス株のうち2株 (A/sw/Obihiro/1/93 と A/sw/Obihiro/2/93) のHA抗原は A/Hokkaido/1/88株に, 残り1株 (A/sw/Obihiro/3/93) は A/Hokkaido/1/93株に類似していることが明らかとなった. HAの塩基置換に基づいて作成した進化系統樹において, A/sw/Obihiro/1/93とA/sw/Obihiro/2/93の2株は1986年から1988年に人から分離された香港型ウイルスと, また, A/sw/Qbihiro/3/93株は1993年に人から分離されたウイルスと同じ集団に属していることが明らかとなった. A/sw/Obihiro/1/93株に対する抗体陽性豚は, 1988年から1994年にかけて, 6.9%から13.8%の割合で認められた. 以上のことから豚は初期の香港型ウイルスと同様に, 最近の人での流行時においても人ウイルスに感染し, 人から感染したウイルスを少なくとも5年間豚集団で保存していることが明らかとなった., 社団法人日本獣医学会, 英語

ウマヘルペスウイルス1型(EHV1)gC糖蛋白の細胞吸着活性
菅原靖広, 松村富夫, 喜田宏, 岡崎克則, 日本獣医学会学術集会講演要旨集, 120th, 113, 1995年09月
日本語

中等毒型ニューカッスル病ウイルス神経馴化株感染鶏の脳病変
Bhaiyat M. I, 古林 与志安, 板倉 智敏, Islam M.A, 喜田 宏, The journal of veterinary medical science, 57, 2, 237, 244, 1995年04月
ウズラより分離した中等毒型ニューカッスル病ウイルス(Q_0)を鶏の脳で10代継代することにより神経馴化株(Q_<10>)を作出し, これらのウイルスをSPF鶏に経鼻接種した. 両ウイルス接種鶏とも接種後1週間は毎日, その後は10, 14および21日後にそれぞれ2羽ずつ剖検し, 病理組織学的およびウイルス学的検索を実施した. (Q_<10>)接種群では, 高い致死率を示し, 神経症状を伴う非化膿性脳炎が所見された. 中枢神経病巣は, parahippocampal cortex, hippocampus, hyperstriatum, neostriatum並びに軟膜下および脳室周囲領域に主座する神経細胞の変性・壊死, 囲管性リンパ球浸潤および巣状ないし瀰慢性アストログリオーシスを特徴としていた. このほか, 神経細胞の変性および軸索スフェロイドの出現を伴う海綿状変化が少数例の脳幹で所見された. 脳のウイルス量は接種後4日目で最大となったが, 接種後6日目以降は皆無であった. (Q_<0>)接種群では, (Q_<10>)接種群とは対照的に, 致死率は0%, 臨床症状も認められず, 脳からウイルスは回収されなかった. これらの中枢神経の組織病変としては, 極く軽微な炎症性変化のみが所見された. 両ウイルス接種鶏間での中枢神経病変のこの様な相違は, ウイルスの神経病原性に関連する..., 社団法人日本獣医学会, 英語

抗血清およびアシクロビルによる新生仔犬の犬ヘルペスウイルス感染症の発病阻止効果
浅野真知, 滝口満喜, 安田準, 橋本晃, 喜田宏, 平井克哉, 日本獣医学会学術集会講演要旨集, 119th, 235, 1995年03月
日本語

インフルエンザウイルスの起源,進化と予測
喜田 宏, 学術月報, 47, 10, p1065, 1071, 1994年10月
日本学術振興会, 日本語

不活化オーエスキー病ウイルスワクチンの経鼻接種によるマウスの感染防御
高田 礼人, 清水 悠紀臣, 喜田 宏, 日本獣医学雑誌, 56, 4, 633, 637, 1994年08月
不活化オーエスキー病ウイルス(ADV)をマウスの鼻腔内に接種することによって, 粘膜局所に抗ウイルスIgAおよびIgG抗体の分泌が誘導された. これらのマウスは鼻腔内への致死量のウイルス攻撃に対し, 完全な感染防御を示した. 不活化ADVにコレラトキシンBサブユニット(CTB)をアジュバントとして混合したワクチンはさらに強い免疫応答を誘導した. 一方, 不活化ADVワクチンを皮下に接種したマウスでは, CTBをアジュバントとして加えた場合でも, 粘膜局所抗体の分泌がほとんど認められなかった. これらのマウスはウイルス攻撃に対してわずかに抵抗性を示したのみで, 半数以上が死亡した. 以上の成績は感染粘膜局所の免疫応答を誘導することによって, 現行の不活化ヘルペスウイルスワクチンでは達成できない初期感染の防御が可能であることを示している. ヘルペスウイルスの初期感染を防御すれば, 潜伏感染およびその後の再活性化の問題も解決されるであろう., 社団法人日本獣医学会, 英語

Pulsed intravenous methylprednisolone combined with oral steroids as the initial treatment of inflammatory myopathies
S. Matsubara, Y. Sawa, M. Takamori, H. Yokoyama, H. Kida, Journal of Neurology, Neurosurgery and Psychiatry, 57, 8, 1008, 1994年
BMJ Publishing Group, 英語, 速報，短報，研究ノート等（学術雑誌）

新型インフルエンザウイルスの出現にかかわる動物とその役割 (野生動物の個体群動態と感染症<特集>)
喜田 宏, 獣医畜産新報, 47, 1, p33, 36, 1994年01月
文永堂出版, 日本語

水禽営巣地におけるインフルエンザウイルスの存続
喜田宏, 伊藤寿啓, 清水悠紀臣, 岡崎克則, 河岡義裕, WEBSTER R G, 日本獣医学会学術集会講演要旨集, 115th, 172, 1993年04月
日本語

新型インフルエンザウイルスの出現機構
喜田 宏, 化学と生物, 31, 3, 154, 162, 1993年03月
学会出版センタ-, 日本語

ESTABLISHMENT OF A FELINE T-LYMPHOBLASTOID CELL-LINE SUSCEPTIBLE TO FELINE IMMUNODEFICIENCY VIRUSES
S FUJIMOTO, M MASUDA, E ONO, H KIDA, Y SHIMIZU, MICROBIOLOGY AND IMMUNOLOGY, 37, 5, 415, 418, 1993年
A feline T-lymphoblastoid cell line susceptible to feline immunodeficiency viruses (FIV) was established and designated as Yu-1 cells. Yu-1 cells continued to grow over one year with more than 100 successive passages in the presence of human recombinant interleukin-2. Surface antigens of Yu-1 cells were feline Pan-T+, CD4+, and CD8-. Susceptibility of Yu-1 cells to FIV strains were higher than that of the primary culture of the feline peripheral blood mononuclear cells, indicating that this cell line should be useful for isolation, propagation, and neutralization test of FIV., CENTER ACADEMIC PUBL JAPAN, 英語, 記事・総説・解説・論説等（学術雑誌）

DETECTION OF FELINE HERPESVIRUS-1 TRANSCRIPTS IN TRIGEMINAL GANGLIA OF LATENTLY INFECTED CATS
Y OHMURA, E ONO, T MATSUURA, H KIDA, Y SHIMIZU, ARCHIVES OF VIROLOGY, 129, 1-4, 341, 347, 1993年
In the trigeminal ganglia of cats latently infected with feline herpesvirus 1, transcripts of opposite strand to the immediate-early mRNA were detected by in situ hybridization and Northern blot analysis., SPRINGER-VERLAG WIEN, 英語, 記事・総説・解説・論説等（学術雑誌）

インフルエンザウイルスの生態--新型ウイルスの出現機構とその予測 (日本ウイルス学会シンポジウム--ウイルス学の進展--1991年)
喜田 宏, ウイルス, 42, 1, 73, 75, 1992年06月
日本ウィルス学会, 日本語

北方圏の水禽営巣地点におけるインフルエンザウイルスおよびパラミクソウイルスの分布‐1991年
伊藤寿啓, 岡崎克則, 河岡義裕, 小野悦郎, 清水悠紀臣, WEBSTER R G, 喜田宏, 日本獣医学会学術集会講演要旨集, 113th, 176, 1992年04月
日本語

豚におけるA型インフルエンザウイルス抗体の最近の動向
後藤 仁, 高井 学, 井口 弘之, Benkele Wilmot, 太田 千佳子, 山本 泰弘, 喜田 宏, 野呂 新一, 桜田 教夫, 日本獣医学雑誌, 54, 2, 235, 241, 1992年04月
1985-90年に採取した4,080例の屠殺豚血清について,各A型インフルエンザウイルスに対する抗体調査をおこなった. 豚型の A/New Jersey/76(HlNl)株に対する抗体陽性豚は, 月によってばらつきがあるが, 観察期間を通じて検出された. 北海道で分離された最近のソ連型ウイルス( HlNl )株に対しても, ヒトでの同型ウイルスの流行後に, それぞれ数か月にわたって抗体陽性豚が観察された. また香港型ウイルス(H3N2)株に対する抗体陽性豚は, 1985-87年間において, ヒトでの同型ウイルスの流行の有無と関係なく検出された. これら3種のウイルス抗体については, これまで我々を含む多くの研究者による調査結果と, 同様の成績が得られた. しかし, 1983-88年分離のH3N2ウイルス株に対する抗体陽性豚は, 1988年にはほとんどみられず,1989年にいたって初めて検出された. さらに, 1989年分離のH3N2ウイルス株に対する抗体陽性豚は, 1988年冬にヒトで同型ウイルスによる激しい流行があったにも拘らず, 1988-90年を通じて検出されないなど, これまでの調査結果と著しく異なる成績が得られた., 社団法人日本獣医学会, 英語

マウス着床前胚に対するセンダイウイルス実験感染の影響
Lavilla Apelo, 喜田 宏, 金川 弘司, 日本獣医学雑誌, 54, 2, 335, 340, 1992年04月
透明帯非除去および除去したマウス桑実期胚を in vitroでセンダイウイルスに感作したのちに一部の胚は感作後48時間培養し in vitroでの発育を, また一部の胚は偽妊娠雌マウス子宮内に移植して in vitroでの発育を検討した. 両群とも, 48時間の培養後に高率に拡張肝盤胞に発育した. 透明帯非除去胚は感染に抵抗して移植後には産子に発育し, 透明帯ウイルス感染の障壁となっていることを示した. これに反して, 透明帯除去胚はウイルスに感染しやすく, 移植した64個のうち僅かに1個の胚が産子に発育したのみであった. 透明帯除去胚の培養液中にウイルスが存在することから, 胚細胞においてウイルス増殖が起っていることが示唆された. 免疫蛍光抗体法により, 透明帯除去胚および透明帯除去胚から発育した胎児の組織にウイルス抗原の存在が確認された. 以上の結果から, 胚細胞におけるセンダイウイルスの増殖が胚の初期発生および胎児への発育を阻害することが示唆された., 社団法人日本獣医学会, 英語