研究者データベース

研究者情報

マスター

アカウント(マスター)

  • 氏名

    苅和 宏明(カリワ ヒロアキ), カリワ ヒロアキ

所属(マスター)

  • 獣医学研究院 獣医学部門 衛生学分野

所属(マスター)

  • 獣医学研究院 獣医学部門 衛生学分野

独自項目

syllabus

  • 2020, 獣医公衆衛生学特論, Advanced Lecture on Veterinary Public Health, 博士後期課程, 獣医学院
  • 2020, 獣医公衆衛生学特論, Advanced Lecture on Veterinary Public Health, 博士後期課程, 国際感染症学院
  • 2020, インターンシップ, Internships, 博士後期課程, 獣医学院
  • 2020, 海外インターンシップA, Internships Abroad A, 博士後期課程, 国際感染症学院, インターンシップ、海外活動、疫学活動、共同研究
  • 2020, 海外インターンシップB, Internships Abroad B, 博士後期課程, 国際感染症学院
  • 2020, 人獣共通感染症対策専門特論, Advanced and Comprehensive Studies on Zoonosis Control, 博士後期課程, 国際感染症学院
  • 2020, 人獣共通感染症学, Zoonotic Science, 学士課程, 獣医学部, 人獣共通感染症、新興・再興感染症、疫学、自然宿主、媒介動物、感染環、ウイルス、細菌、原虫、寄生虫 
  • 2020, 畜産食品衛生学, Milk and Meat Hygiene, 学士課程, 農学部, 畜産食品、乳、肉、食品衛生、食中毒
  • 2020, 獣医公衆衛生学, Veterinary Public Health, 学士課程, 獣医学部, 獣医衛生, 国民衛生の動向, 獣医疫学, 感染症・非感染症の疫学, 感染症の予防と防疫, 環境衛生と環境汚染
  • 2020, 獣医公衆衛生学実習, Practice in Veterinary Public Health, 学士課程, 獣医学部, 環境衛生と環境汚染, 大気の衛生, 上水・下水・汚水, 水質検査、感染症、人獣共通感染症

timetable

  • 博士後期課程, 国際感染症学院, 2020, 感染症学特別研究Ⅰ
  • 博士後期課程, 国際感染症学院, 2020, 感染症学特別演習
  • 博士後期課程, 国際感染症学院, 2020, 感染症学特別研究ⅡA
  • 博士後期課程, 国際感染症学院, 2020, 感染症学特別研究ⅡB
  • 博士後期課程, 国際感染症学院, 2020, 海外インターンシップA
  • 博士後期課程, 国際感染症学院, 2020, 海外インターンシップB
  • 博士後期課程, 獣医学研究科, 2020, 獣医科学特別研究
  • 博士後期課程, 獣医学研究科, 2020, 人獣共通感染症対策専門特論
  • 博士後期課程, 獣医学研究科, 2020, 獣医科学特論演習
  • 博士後期課程, 獣医学研究科, 2020, 獣医科学基礎科目B 獣医公衆衛生学特論
  • 博士後期課程, 獣医学院, 2020, インターンシップ
  • 学士課程, 獣医学部, 2020, 獣医公衆衛生学
  • 学士課程, 獣医学部, 2020, 獣医公衆衛生学実習
  • 学士課程, 獣医学部, 2020, 環境衛生学
  • 学士課程, 獣医学部, 2020, 人獣共通感染症学
  • 学士課程, 獣医学部, 2020, 研究・臨床セミナー
  • 学士課程, 獣医学部, 2020, アドバンスト演習

researchmap

プロフィール情報

学位

  • 博士(獣医学)(北海道大学)

プロフィール情報

  • 苅和, KARIWA
  • 宏明, Hiroaki
  • ID各種

    200901056681540702

対象リソース

業績リスト

研究キーワード

  • 人獣共通感染症   ウイルス学   公衆衛生学   Zoonosis   Virology   Public Health   

研究分野

  • ライフサイエンス / 獣医学
  • ライフサイエンス / 獣医学
  • ライフサイエンス / 獣医学

受賞

  • 2007年04月 日本獣医学会 日本獣医学会賞
     野生げっ歯類を対象としたハンタウイルス感染症の比較疫学的研究 
    受賞者: 苅和 宏明

論文

  • Kentaro Uemura, Haruaki Nobori, Akihiko Sato, Takao Sanaki, Shinsuke Toba, Michihito Sasaki, Akiho Murai, Noriko Saito-Tarashima, Noriaki Minakawa, Yasuko Orba, Hiroaki Kariwa, William W Hall, Hirofumi Sawa, Akira Matsuda, Katsumi Maenaka
    iScience 24 10 103120 - 103120 2021年10月22日 
    Newly emerging or re-emerging viral infections continue to cause significant morbidity and mortality every year worldwide, resulting in serious effects on both health and the global economy. Despite significant drug discovery research against dengue viruses (DENVs) and severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), no fully effective and specific drugs directed against these viruses have been discovered. Here, we examined the anti-DENV activity of tubercidin derivatives from a compound library from Hokkaido University and demonstrated that 5-hydroxymethyltubercidin (HMTU, HUP1108) possessed both potent anti-flavivirus and anti-coronavirus activities at submicromolar levels without significant cytotoxicity. Furthermore, HMTU inhibited viral RNA replication and specifically inhibited replication at the late stages of the SARS-CoV-2 infection process. Finally, we demonstrated that HMTU 5'-triphosphate inhibited RNA extension catalyzed by the viral RNA-dependent RNA polymerase. Our findings suggest that HMTU has the potential of serving as a lead compound for the development of a broad spectrum of antiviral agents, including SARS-CoV-2.
  • Fumihiro Kodama, Hiroki Yamaguchi, Eunsil Park, Kango Tatemoto, Mariko Sashika, Ryo Nakao, Yurino Terauchi, Keita Mizuma, Yasuko Orba, Hiroaki Kariwa, Katsuro Hagiwara, Katsunori Okazaki, Akiko Goto, Rika Komagome, Masahiro Miyoshi, Takuya Ito, Kimiaki Yamano, Kentaro Yoshii, Chiaki Funaki, Mariko Ishizuka, Asako Shigeno, Yukari Itakura, Lesley Bell-Sakyi, Shunji Edagawa, Atsushi Nagasaka, Yoshihiro Sakoda, Hirofumi Sawa, Ken Maeda, Masayuki Saijo, Keita Matsuno
    Nature communications 12 1 5539 - 5539 2021年09月20日 
    The increasing burden of tick-borne orthonairovirus infections, such as Crimean-Congo hemorrhagic fever, is becoming a global concern for public health. In the present study, we identify a novel orthonairovirus, designated Yezo virus (YEZV), from two patients showing acute febrile illness with thrombocytopenia and leukopenia after tick bite in Hokkaido, Japan, in 2019 and 2020, respectively. YEZV is phylogenetically grouped with Sulina virus detected in Ixodes ricinus ticks in Romania. YEZV infection has been confirmed in seven patients from 2014-2020, four of whom were co-infected with Borrelia spp. Antibodies to YEZV are found in wild deer and raccoons, and YEZV RNAs have been detected in ticks from Hokkaido. In this work, we demonstrate that YEZV is highly likely to be the causative pathogen of febrile illness, representing the first report of an endemic infection associated with an orthonairovirus potentially transmitted by ticks in Japan.
  • Inactivation of SARS-CoV-2 by povidone-iodine products: implications for effective mouth rinsing and gargling
    Hiroaki Kariwa, Hirofumi Sawa, Shintaro Kobayashi
    The Japanese Journal of Veterinary Research 69 3 183 - 187 2021年09月 [査読有り][通常論文]
  • Takuma Izumi, Yuhei Morioka, Syun-Ichi Urayama, Daisuke Motooka, Tomokazu Tamura, Takahiro Kawagishi, Yuta Kanai, Takeshi Kobayashi, Chikako Ono, Akinari Morinaga, Takahiro Tomiyama, Norifumi Iseda, Yukiko Kosai, Shoichi Inokuchi, Shota Nakamura, Tomohisa Tanaka, Kohji Moriishi, Hiroaki Kariwa, Tomoharu Yoshizumi, Masaki Mori, Yoshiharu Matsuura, Takasuke Fukuhara
    Viruses 13 7 2021年07月06日 
    Although viruses infect various organs and are associated with diseases, there may be many unidentified pathogenic viruses. The recent development of next-generation sequencing technologies has facilitated the establishment of an environmental viral metagenomic approach targeting the intracellular viral genome. However, an efficient method for the detection of a viral genome derived from an RNA virus in animal or human samples has not been established. Here, we established a method for the efficient detection of RNA viruses in human clinical samples. We then tested the efficiency of the method compared to other conventional methods by using tissue samples collected from 57 recipients of living donor liver transplantations performed between June 2017 and February 2019 at Kyushu University Hospital. The viral read ratio in human clinical samples was higher by the new method than by the other conventional methods. In addition, the new method correctly identified viral RNA from liver tissues infected with hepatitis C virus. This new technique will be an effective tool for intracellular RNA virus surveillance in human clinical samples and may be useful for the detection of new RNA viruses associated with diseases.
  • Keisuke Maezono, Shintaro Kobayashi, Koshiro Tabata, Kentaro Yoshii, Hiroaki Kariwa
    Scientific reports 11 1 9213 - 9213 2021年04月28日 
    West Nile virus (WNV), a member of the Japanese encephalitis virus (JEV) serocomplex group, causes lethal encephalitis in humans and horses. Because serodiagnosis of WNV and JEV is hampered by cross-reactivity, the development of a simple, secure, and WNV-specific serodiagnostic system is required. The coexpression of prM protein and E protein leads to the secretion of subviral particles (SPs). Deletion of the C-terminal region of E protein is reported to affect the production of SPs by some flaviviruses. However, the influence of such a deletion on the properties and antigenicity of WNV E protein is unclear. We analyzed the properties of full-length E protein and E proteins lacking the C-terminal region as novel serodiagnostics for WNV infection. Deletion of the C-terminal region of E protein suppressed the formation of SPs but did not affect the production of E protein. The sensitivity of an enzyme-linked immunosorbent assay (ELISA) using the full-length E protein was higher than that using the truncated E proteins. Furthermore, in the ELISA using full-length E protein, there was little cross-reactivity with anti-JEV antibodies, and the sensitivity was similar to that of the neutralization test.
  • Mai Kishimoto, Kentaro Uemura, Takao Sanaki, Akihiko Sato, William W Hall, Hiroaki Kariwa, Yasuko Orba, Hirofumi Sawa, Michihito Sasaki
    Viruses 13 3 2021年02月28日 
    Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) utilizes host proteases, including a plasma membrane-associated transmembrane protease, serine 2 (TMPRSS2) to cleave and activate the virus spike protein to facilitate cellular entry. Although TMPRSS2 is a well-characterized type II transmembrane serine protease (TTSP), the role of other TTSPs on the replication of SARS-CoV-2 remains to be elucidated. Here, we have screened 12 TTSPs using human angiotensin-converting enzyme 2-expressing HEK293T (293T-ACE2) cells and Vero E6 cells and demonstrated that exogenous expression of TMPRSS11D and TMPRSS13 enhanced cellular uptake and subsequent replication of SARS-CoV-2. In addition, SARS-CoV-1 and SARS-CoV-2 share the same TTSPs in the viral entry process. Our study demonstrates the impact of host TTSPs on infection of SARS-CoV-2, which may have implications for cell and tissue tropism, for pathogenicity, and potentially for vaccine development.
  • Yuji Takahashi, Shintaro Kobayashi, Mariko Ishizuka, Minato Hirano, Memi Muto, Shoko Nishiyama, Hiroaki Kariwa, Kentaro Yoshii
    The Journal of general virology 101 5 497 - 509 2020年05月 [査読有り][通常論文]
     
    Tick-borne encephalitis virus (TBEV) is a zoonotic virus in the genus Flavivirus, family Flaviviridae. TBEV is widely distributed in northern regions of the Eurasian continent, including Japan, and causes severe encephalitis in humans. Tick-borne encephalitis (TBE) was recently reported in central Hokkaido, and wild animals with anti-TBEV antibodies were detected over a wide area of Hokkaido, although TBEV was only isolated in southern Hokkaido. In this study, we conducted a survey of ticks to isolate TBEV in central Hokkaido. One strain, designated Sapporo-17-Io1, was isolated from ticks (Ixodes ovatus) collected in Sapporo city. Sequence analysis revealed that the isolated strain belonged to the Far Eastern subtype of TBEV and was classified in a different subcluster from Oshima 5-10, which had previously been isolated in southern Hokkaido. Sapporo-17-Io1 showed similar growth properties to those of Oshima 5-10 in cultured cells and mouse brains. The mortality rate of mice infected intracerebrally with each virus was similar, but the survival time of mice inoculated with Sapporo-17-Io1 was significantly longer than that of mice inoculated with Oshima 5-10. These results indicate that the neurovirulence of Sapporo-17-Io1 was lower than that of Oshima 5-10. Using an infectious cDNA clone, the replacement of genes encoding non-structural genes from Oshima 5-10 with those from Sapporo-17-Io1 attenuated the neuropathogenicity of the cloned viruses. This result indicated that the non-structural proteins determine the neurovirulence of these two strains. Our results provide important insights for evaluating epidemiological risk in TBE-endemic areas of Hokkaido.
  • Shintaro Kobayashi, Chisato Kaneko, Ryoko Kawakami, Rie Hasebe, Hirofumi Sawa, Kentaro Yoshii, Hiroaki Kariwa
    Scientific reports 10 1 7168 - 7168 2020年04月28日 [査読有り][通常論文]
     
    West Nile virus (WNV) is an important cause of viral encephalitis in birds and animals, including humans. Amino acid 159 of the envelope (E) protein is reportedly implicated in the different levels of neurovirulence in mice infected with WNV NY99 or Eg101. We investigated the role of amino acid 159 of the E protein in the pathogenesis of WNV infection. We produced recombinant WNV with the structural proteins of the NY99 or Eg101 strain (NY-WT or EgCME-WT) and mutant viruses with substitutions of amino acid 159 of the E protein (NY-E-V159I or EgCME-E-I159V). The NY-WT and NY-E-V159I or EgCME-WT and EgCME-E-I159V titers in culture supernatant were similar. The mortality rate and viral titer in the brains of mice inoculated intraperitoneally with NY-WT or NY-E-V159I were also similar. In contrast, the mortality rate and viral titer in the brains of mice inoculated intracranially with EgCME-E-I159V were significantly higher than those of mice inoculated with EgCME-WT. The numbers of CD3-positive and CD8-positive T cells were greater in brains inoculated with EgCME-E-I159V than in those inoculated with EgCME-WT. Therefore, amino acid 159 of the E protein modulates the pathogenicity of WNV by affecting viral replication and T-cell infiltration in the brain.
  • Jan Clement, Clas Ahlm, Tatjana Avšič-Županc, Jason Botten, Kartik Chandran, Colleen B Jonsson, Hiroaki Kariwa, Jonas Klingström, Boris Klempa, Detlev H Krüger, Herwig Leirs, Dexin Li, Mifang Liang, Alemka Markotić, Anna Papa, Connie S Schmaljohn, Nicole D Tischler, Rainer G Ulrich, Antti Vaheri, Cecilia Vial, Richard Yanagihara, Piet Maes
    Antiviral research 176 104733 - 104733 2020年04月 
    The 2019 11th International Conference on Hantaviruses (ICH 2019) was organized by the International Society for Hantaviruses (ISH), and held on September 1-4, 2019, at the Irish College, in Leuven, Belgium. These ICHs have been held every three years since 1989. ICH 2019 was attended by 158 participants from 33 countries. The current report summarizes research presented on all aspects of hantavirology: ecology; pathogenesis and immune responses; virus phylogeny, replication and morphogenesis; epidemiology; vaccines, therapeutics and prevention; and clinical aspects and diagnosis.
  • Shintaro Kobayashi, Kentaro Yoshii, Wallaya Phongphaew, Memi Muto, Minato Hirano, Yasuko Orba, Hirofumi Sawa, Hiroaki Kariwa
    PLoS pathogens 16 1 e1008238  2020年01月 [査読有り][通常論文]
     
    West Nile virus (WNV) belongs to the Flaviviridae family and has emerged as a significant cause of viral encephalitis in birds and animals including humans. WNV replication directly induces neuronal injury, followed by neuronal cell death. We previously showed that accumulation of ubiquitinated protein aggregates was involved in neuronal cell death in the WNV-infected mouse brain. In this study, we attempted to elucidate the mechanisms of the accumulation of protein aggregates in the WNV-infected cells. To identify the viral factor inducing the accumulation of ubiquitinated proteins, intracellular accumulation of ubiquitinated proteins was examined in the cells expressing the viral protein. Expression of capsid (C) protein induced the accumulation, while mutations at residues L51 and A52 in C protein abrogated the accumulation. Wild-type (WT) or mutant WNV in which mutations were introduced into the residues was inoculated into human neuroblastoma cells. The expression levels of LC3-II, an autophagy-related protein, and AMP-activated protein kinase (AMPK), an autophagy inducer, were reduced in the cells infected with WT WNV, while the reduction was not observed in the cells infected with WNV with the mutations in C protein. Similarly, ubiquitination and degradation of AMPK were only observed in the cells infected with WT WNV. In the cells expressing C protein, AMPK was co-precipitated with C protein and mutations in L51 and A52 reduced the interaction. Although the viral replication was not affected, the accumulation of ubiquitinated proteins in brain and neurological symptoms were attenuated in the mouse inoculated with WNV with the mutations in C protein as compared with that with WT WNV. Taken together, ubiquitination and degradation of AMPK by C protein resulted in the inhibition of autophagy and the accumulation of protein aggregates, which contributes to the development of neurological disease.
  • Christida E Wastika, Michihito Sasaki, Kentaro Yoshii, Paulina D Anindita, Bernard M Hang'ombe, Aaron S Mweene, Shintaro Kobayashi, Hiroaki Kariwa, Michael J Carr, William W Hall, Yuki Eshita, Yasuko Orba, Hirofumi Sawa
    Archives of virology 164 8 2165 - 2170 2019年08月 [査読有り][通常論文]
     
    Zika virus (ZIKV) circulation occurs between non-human primates (NHPs) in a sylvatic transmission cycle. To investigate evidence of flavivirus infection in NHPs in Zambia, we performed a plaque reduction neutralization test (PRNT) to quantify neutralizing antibodies. PRNT revealed that sera from NHPs (African green monkeys and baboons) exhibited neutralizing activity against ZIKV (34.4%; 33/96), whereas a PRNT for yellow fever virus using NHP sera showed no neutralization activity. ZIKV genomic RNA was not detected in splenic tissues from NHPs, suggesting that the presence of anti-ZIKV neutralizing antibodies represented resolved infections. Our evidence suggests that ZIKV is maintained in NHP reservoirs in Zambia.
  • Jamsransuren, Dulamjav, Yoshii, Kentaro, Kariwa, Hiroaki, Asakawa, Mitsuhiko, Okuda, Kei, Fujii, Kei, Fukumoto, Shinya, Umemiya-Shirafuji, Rika, Sasaki, Motoki, Matsumoto, Kotaro, Yamaguchi, Emi, Ogawa, Haruko, Imai, Kunitoshi
    Japanese Journal of Veterinary Research 67 2 163 - 172 2019年05月 [査読有り][通常論文]
  • Tapiwa Lundu, Kentaro Yoshii, Shintaro Kobayashi, Shigeru Morikawa, Toshio Tsubota, Naoaki Misawa, Daisuke Hayasaka, Hiroaki Kariwa
    Japanese Journal of Veterinary Research 66 1 21 - 28 2018年 [査読有り][通常論文]
     
    Severe fever with thrombocytopenia syndrome (SFTS) is a newly recognized zoonosis that occurs in China, Japan, and South Korea and is caused by the SFTS virus (SFTSV), which is in the genus Phlebovirus, family Phenuiviridae. Since its discovery in Japan in 2013, SFTS has been reported in the western parts of the country. To elucidate the distribution of SFTSV, we conducted a serological survey of deer and rodents. Serum was screened using enzyme-linked immunosorbent assay (ELISA) and suspected cases were further tested with an indirect immunofluorescence antibody (IFA) assay. Serum samples from 315 deer from Hokkaido (non-endemic area), 41 deer from Miyazaki (endemic area), and 910 rodents from six locations in Japan were tested. Of the 41 deer from Miyazaki, 2 (4.9%) had high ELISA optical density (OD) values (0.1 < OD < 0.3) and a positive IFA result. All of the deer samples from Hokkaido were negative by ELISA (OD < 0.1). No SFTSV-positive rodents were found. Our results indicate that deer in Miyazaki were exposed to SFTSV, unlike deer from Hokkaido (P < 0.05).
  • Tapiwa Lundu, Yoshimi Tsuda, Ryo Ito, Kenta Shimizu, Shintaro Kobayashi, Kentaro Yoshii, Kumiko Yoshimatsu, Jiro Arikawa, Hiroaki Kariwa
    Biomedical research (Tokyo, Japan) 39 1 27 - 38 2018年 [査読有り][通常論文]
     
    Severe fever with thrombocytopenia syndrome phlebovirus (SFTSV) is a newly emerged phlebovirus identified in China, Japan, and South Korea. Phlebovirus glycoproteins (GP) play a key role in targeting viral structural components to the budding compartments in the ER-Golgi intermediate compartment (ERGIC) and Golgi complex. However, the role of SFTSV GP in targeting structural proteins to the ERGIC and Golgi complex remains unresolved. In this study, we show that SFTSV GP plays a significant role in targeting RNA-dependent RNA polymerase (L) and nucleocapsid protein (NP) to the budding sites. Confocal microscopy was used to investigate the subcellular localization of SFTSV structural proteins. In SFTSV-infected cells, GP and L localized to the ER, ERGIC and Golgi complex, whereas NP localized to the ERGIC and Golgi complex. In addition, GP colocalized with L and NP in infected cells. In cells singly transfected with GP, L or NP, GP localized to the same subcellular compartments as in infected cells. However, L or NP alone did not localize to the ER, ERGIC, or Golgi complex. Cotransfection experiments showed that GP altered the localization of L to the ERGIC and Golgi complex but not that of NP. Interestingly, plasmid-expressed NP fused with a hemagglutinin tag localized to the ERGIC and Golgi complex when expressed in SFTSV-infected cells and colocalised with GP, suggesting that GP plays a role in the subcellular localization of L and NP in infected cells. Thus, the SFTSV structural components start to assemble at the ERGIC to Golgi complex. GP is required for transporting L and NP to the ERGIC and Golgi complex. In addition, targeting of NP requires interaction with other factors besides GP.
  • Ludek Eyer, Hirofumi Kondo, Darina Zouharova, Minato Hirano, James J Valdés, Memi Muto, Tomas Kastl, Shintaro Kobayashi, Jan Haviernik, Manabu Igarashi, Hiroaki Kariwa, Marketa Vaculovicova, Jiri Cerny, Rene Kizek, Andrea Kröger, Stefan Lienenklaus, Milan Dejmek, Radim Nencka, Martin Palus, Jiri Salat, Erik De Clercq, Kentaro Yoshii, Daniel Ruzek
    Journal of virology 91 21 2017年11月01日 [査読有り][通常論文]
     
    Tick-borne encephalitis virus (TBEV) causes a severe and potentially fatal neuroinfection in humans. Despite its high medical relevance, no specific antiviral therapy is currently available. Here we demonstrate that treatment with a nucleoside analog, 7-deaza-2'-C-methyladenosine (7-deaza-2'-CMA), substantially improved disease outcomes, increased survival, and reduced signs of neuroinfection and viral titers in the brains of mice infected with a lethal dose of TBEV. To investigate the mechanism of action of 7-deaza-2'-CMA, two drug-resistant TBEV clones were generated and characterized. The two clones shared a signature amino acid substitution, S603T, in the viral NS5 RNA-dependent RNA polymerase (RdRp) domain. This mutation conferred resistance to various 2'-C-methylated nucleoside derivatives, but no cross-resistance was seen with other nucleoside analogs, such as 4'-C-azidocytidine and 2'-deoxy-2'-beta-hydroxy-4'-azidocytidine (RO-9187). All-atom molecular dynamics simulations revealed that the S603T RdRp mutant repels a water molecule that coordinates the position of a metal ion cofactor as 2'-C-methylated nucleoside analogs approach the active site. To investigate its phenotype, the S603T mutation was introduced into a recombinant TBEV strain (Oshima-IC) generated from an infectious cDNA clone and into a TBEV replicon that expresses a reporter luciferase gene (Oshima-REP-luc2A). The mutants were replication impaired, showing reduced growth and a small plaque size in mammalian cell culture and reduced levels of neuroinvasiveness and neurovirulence in rodent models. These results indicate that TBEV resistance to 2'-C-methylated nucleoside inhibitors is conferred by a single conservative mutation that causes a subtle atomic effect within the active site of the viral NS5 RdRp and is associated with strong attenuation of the virus.IMPORTANCE This study found that the nucleoside analog 7-deaza-2'-C-methyladenosine (7-deaza-2'-CMA) has high antiviral activity against tick-borne encephalitis virus (TBEV), a pathogen that causes severe human neuroinfections in large areas of Europe and Asia and for which there is currently no specific therapy. Treating mice infected with a lethal dose of TBEV with 7-deaza-2'-CMA resulted in significantly higher survival rates and reduced the severity of neurological signs of the disease. Thus, this compound shows promise for further development as an anti-TBEV drug. It is important to generate drug-resistant mutants to understand how the drug works and to develop guidelines for patient treatment. We generated TBEV mutants that were resistant not only to 7-deaza-2'-CMA but also to a broad range of other 2'-C-methylated antiviral medications. Our findings suggest that combination therapy may be used to improve treatment and reduce the emergence of drug-resistant viruses during nucleoside analog therapy for TBEV infection.
  • Minato Hirano, Memi Muto, Mizuki Sakai, Hirofumi Kondo, Shintaro Kobayashi, Hiroaki Kariwa, Kentaro Yoshii
    Proceedings of the National Academy of Sciences of the United States of America 114 37 9960 - 9965 2017年09月12日 [査読有り][通常論文]
     
    Neurological diseases caused by encephalitic flaviviruses are severe and associated with high levels of mortality. However, little is known about the detailed mechanisms of viral replication and pathogenicity in the brain. Previously, we reported that the genomic RNA of tick-borne encephalitis virus (TBEV), a member of the genus Flavivirus, is transported and replicated in the dendrites of neurons. In the present study, we analyzed the transport mechanism of the viral genome to dendrites. We identified specific sequences of the 5' untranslated region of TBEV genomic RNA that act as a cis-acting element for RNA transport. Mutated TBEV with impaired RNA transport in dendrites caused a reduction in neurological symptoms in infected mice. We show that neuronal granules, which regulate the transport and local translation of dendritic mRNAs, are involved in TBEV genomic RNA transport. TBEV genomic RNA bound an RNA-binding protein of neuronal granules and disturbed the transport of dendritic mRNAs. These results demonstrated a neuropathogenic virus hijacking the neuronal granule system for the transport of viral genomic RNA in dendrites, resulting in severe neurological disease.
  • Shintaro Kobayashi, Kentaro Yoshii, Minato Hirano, Memi Muto, Hiroaki Kariwa
    Journal of virological methods 240 14 - 20 2017年02月 [査読有り][通常論文]
     
    Reverse genetics systems facilitate investigation of many aspects of the life cycle and pathogenesis of viruses. However, genetic instability in Escherichia coli has hampered development of a reverse genetics system for West Nile virus (WNV). In this study, we developed a novel reverse genetics system for WNV based on homologous recombination in mammalian cells. Introduction of the DNA fragment coding for the WNV structural protein together with a DNA-based replicon resulted in the release of infectious WNV. The growth rate and plaque size of the recombinant virus were almost identical to those of the parent WNV. Furthermore, chimeric WNV was produced by introducing the DNA fragment coding for the structural protein and replicon plasmid derived from various strains. Here, we report development of a novel system that will facilitate research into WNV infection.
  • Hiroaki Kariwa
    Uirusu 67 1 25 - 32 2017年 [査読有り][通常論文]
     
    Hantaviruses belongs to the genus Hantavirus in the family Bunyaviridae are maintained in rodents and infects to humans by inhalation of the aerosol of infected rodent excreta. In this article, the epidemiology of hantavirus infection and the special relationship between rodent and hantavirus are described. Hantavirus infections include hemorrhagic fever with renal syndrome (HFRS) and hantavirus cardiopulmonary syndrome (HCPS). HFRS is characterized high fever, hemorrhage, and renal disorder. HFRS is distributed in East Asia, Europe, and Russia. While HCPS is characterized acute respiratory dysfunction and cardiogenic shock. The distribution of HCPS is limited in North and South Americas. In Japan's neighboring countries, such as Russia, China, and Korea, large numbers of HFRS patients are reported in association with multiple hantaviruses. In Japan, hantavirus infection has not been reported since 1985 but grey red-backed vole (Myodes rufocanus bedfordiae) inhabiting Hokkaido maintain one of the hantaviruses. Coevolution between hantavirus and host may have been occurred during a long period. The endemic areas of hantavirus infection are strongly associated with the distribution of host animal carrying pathogenic hantaviruses.
  • Targeting of severe fever with thrombocytopenia syndrome virus structural proteins to the ERGIC (endoplasmic reticulum Golgi intermediate compartment) and Golgi complex.
    Lundu T, Tsuda Y, Ito R, Shimizu K, Kobayashi S, Yoshii K, Yoshimatsu K, Arikawa J, Kariwa H
    Biomed Res-Tokyo 78 8 1453 - 1460 2017年 [査読有り][通常論文]
  • HokkaidoウイルスとPuumalaウイルスの遺伝子再集合体の作出とその性状解析
    岩崎 里菜, 真田 崇弘, 好井 健太朗, 苅和 宏明
    日本獣医学会学術集会講演要旨集 157回 439 - 439 (公社)日本獣医学会 2014年08月 [査読有り][通常論文]
  • 初代培養マウス脳細胞を用いた脳炎フラビウイルスの増殖機構の解析
    平野 港, 好井 健太朗, 境 瑞樹, 長谷部 理絵, 苅和 宏明
    日本獣医学会学術集会講演要旨集 156回 302 - 302 (公社)日本獣医学会 2013年08月 [査読有り][通常論文]
  • 極東型ダニ媒介性脳炎ウイルスの強毒化に関わるウイルス側因子の特定
    境 瑞紀, 好井 健太朗, 寸田 祐嗣, 横澤 香菜, 平野 港, 苅和 宏明
    日本獣医学会学術集会講演要旨集 156回 303 - 303 (公社)日本獣医学会 2013年08月 [査読有り][通常論文]
  • 新世界ハンタウイルス感染のためのハンタウイルス組み換え核蛋白を用いた血清型鑑別ELISA法の開発
    駒 貴明, 吉松 組子, 垂石 みどり, 宮下 大輔, 遠藤 理香, 清水 健太, 安田 俊平, 天田 貴子, 瀬戸 隆, 村田 亮, 吉田 喜香, 苅和 宏明, 高島 郁夫, 有川 二郎
    北海道医学雑誌 88 1 35 - 35 2013年01月 [査読無し][通常論文]
  • ダニ媒介性フラビウイルスによる中枢神経系障害に関わるウイルス因子の同定
    好井 健太朗, 寸田 祐嗣, 境 瑞紀, 苅和 宏明, Holbrook Michael, 高島 郁夫
    日本獣医学会学術集会講演要旨集 154回 264 - 264 (公社)日本獣医学会 2012年08月 [査読有り][通常論文]
  • 極東型ダニ媒介性脳炎ウイルスの強毒化に関わるウイルス側因子の特定
    境 瑞紀, 好井 健太朗, 横澤 香菜, 苅和 宏明
    日本獣医学会学術集会講演要旨集 154回 264 - 264 (公社)日本獣医学会 2012年08月 [査読有り][通常論文]
  • ダニ媒介性フラビウイルスのインターフェロンアンタゴニスト作用の解析
    鶴田 征太郎, 好井 健太朗, 境 瑞紀, 苅和 宏明
    日本獣医学会学術集会講演要旨集 154回 264 - 264 (公社)日本獣医学会 2012年08月 [査読有り][通常論文]
  • 新たに分離されたHokkaidoウイルスの性状解析
    真田 崇弘, 尾崎 由佳, 瀬戸 隆弘, 中尾 桃子, Saasa Ngonda, 吉松 組子, 有川 二郎, 好井 健太朗, 苅和 宏明
    日本獣医学会学術集会講演要旨集 154回 265 - 265 (公社)日本獣医学会 2012年08月 [査読有り][通常論文]
  • 好井健太朗, 山崎翔子, 持舘景太, 苅和宏明, 高島郁夫
    獣医畜産新報 1090 377-378  2012年05月01日 [査読無し][通常論文]
  • 好井 健太朗, 持舘 景太, 苅和 宏明
    獣医畜産新報 64 10 801 - 803 文永堂出版 2011年10月 [査読無し][通常論文]
  • ダニ媒介性脳炎ウイルスのE蛋白糖鎖は哺乳動物細胞におけるウイルス粒子分泌に影響する
    柳原 なつみ, 好井 健太朗, 後藤 明子, 伊川 綾恵, 石塚 万里子, 苅和 宏明, 高島 郁夫
    北海道獣医師会雑誌 55 8 415 - 415 (公社)北海道獣医師会 2011年08月 [査読有り][通常論文]
  • 2008年北海道で分離されたダニ媒介性脳炎ウイルスOshima 08-AS株の病原性解析
    山崎 翔子, 好井 健太朗, 真田 崇弘, 苅和 宏明, 高島 郁夫
    北海道獣医師会雑誌 55 8 416 - 416 (公社)北海道獣医師会 2011年08月 [査読有り][通常論文]
  • 2008年北海道で分離されたダニ媒介性脳炎ウイルスOshima 08-AS株の病原性解析
    山崎 翔子, 好井 健太朗, 真田 崇弘, 苅和 宏明, 高島 郁夫
    日本獣医学会学術集会講演要旨集 152回 255 - 255 (公社)日本獣医学会 2011年08月 [査読有り][通常論文]
  • 野生マウス由来Oas遺伝子座導入コンジェニックマウスにおけるダニ媒介性脳炎ウイルスの神経病原性の解析
    好井 健太朗, 森藤 可南子, 永田 典代, 佐々木 宣哉, 苅和 宏明, 安居院 高志, 高島 郁夫
    日本獣医学会学術集会講演要旨集 152回 256 - 256 (公社)日本獣医学会 2011年08月 [査読有り][通常論文]
  • ダニ媒介性脳炎ウイルスのE蛋白糖鎖は哺乳動物細胞におけるウイルス粒子分泌に影響する
    柳原 なつみ, 好井 健太朗, 石塚 万里子, 苅和 宏明, 高島 郁夫
    日本獣医学会学術集会講演要旨集 152回 256 - 256 (公社)日本獣医学会 2011年08月 [査読有り][通常論文]
  • エゾヤチネズミ(Myodes rufocanus)の腎臓由来細胞系の確立とHokkaidoウイルス分離への応用
    真田 崇弘, 瀬戸 隆弘, 尾崎 由佳, Ngonda Saasa, 好井 健太朗, 苅和 宏明
    日本獣医学会学術集会講演要旨集 152回 256 - 256 (公社)日本獣医学会 2011年08月 [査読有り][通常論文]
  • 苅和 宏明, 好井 健太朗, 高島 郁夫
    公衆衛生 75 1 36 - 38 医学書院 2011年01月 [査読無し][通常論文]
  • 2008年北海道におけるダニ媒介性脳炎ウイルスの分離と性状解析
    山崎 翔子, 好井 健太朗, 持舘 景太, 村田 亮, 真田 崇弘, 苅和 宏明, 高島 郁夫
    日本獣医学会学術集会講演要旨集 150回 247 - 247 (公社)日本獣医学会 2010年09月 [査読有り][通常論文]
  • ダニ媒介性脳炎ウイルスのE蛋白の糖鎖付加がウイルス性状に与える影響
    柳原 なつみ, 好井 健太朗, 後藤 明子, 伊川 綾恵, 石塚 万里子, 苅和 宏明, 高島 郁夫
    日本獣医学会学術集会講演要旨集 150回 248 - 248 (公社)日本獣医学会 2010年09月 [査読有り][通常論文]
  • ダニ媒介性脳炎ウイルスSofjin株の感染性cDNAの構築
    高野 絢子, 大森 優紀, 好井 健太朗, 横澤 香菜, 苅和 宏明, 高島 郁夫
    日本獣医学会学術集会講演要旨集 150回 248 - 248 (公社)日本獣医学会 2010年09月 [査読有り][通常論文]
  • ダニ媒介性脳炎/オムスク出血熱のキメラウイルスの作成と性状解析
    好井 健太朗, 寸田 祐嗣, 苅和 宏明, Holbrook Michael, 高島 郁夫
    日本獣医学会学術集会講演要旨集 150回 248 - 248 (公社)日本獣医学会 2010年09月 [査読有り][通常論文]
  • メキシコ由来のハンタウイルスに対するモノクローナル抗体の作出と各種ハンタウイルスに対する反応性の検討
    吉田 喜香, 苅和 宏明, 真田 崇弘, Saasa Ngonda, 瀬戸 隆弘, 吉松 組子, 有川 二郎, 好井 健太朗, 高島 郁夫
    日本獣医学会学術集会講演要旨集 150回 249 - 249 (公社)日本獣医学会 2010年09月 [査読有り][通常論文]
  • Puumalaウイルスを感染させたシリアンハムスター(Mesocricetus auratus)の感染動態の解析
    真田 崇弘, 苅和 宏明, 谷川 洋一, Nur Hardy Abu Daud, 瀬戸 隆弘, 永田 典代, 吉松 組子, 有川 二郎, 好井 健太朗, 高島 郁夫
    日本獣医学会学術集会講演要旨集 150回 252 - 252 (公社)日本獣医学会 2010年09月 [査読有り][通常論文]
  • 村田亮, 好井健太朗, 苅和宏明, 高島郁夫
    獣医畜産新報 1064 208-209  2010年03月01日 [査読無し][通常論文]
  • 好井健太朗, 苅和宏明, 高島郁夫
    北海道獣医師会雑誌 54 1 2-7  2010年01月10日 [査読無し][通常論文]
  • 極東ロシアの野鳥におけるウエストナイル熱の血清疫学調査と中和試験の評価
    村田 亮, 橋口 和明, 好井 健太朗, 野田 寛, 伊川 綾恵, 原田 祐里, 苅和 宏明, 高島 郁夫
    日本獣医学会学術集会講演要旨集 148回 236 - 236 (公社)日本獣医学会 2009年09月 [査読有り][通常論文]
  • ダニ媒介性脳炎ウイルスSofjin株のレプリコンの構築
    高野 絢子, 大森 優紀, 好井 健太朗, 石塚 万里子, 村田 亮, 苅和 宏明, 高島 郁夫
    日本獣医学会学術集会講演要旨集 148回 236 - 236 (公社)日本獣医学会 2009年09月 [査読有り][通常論文]
  • オムスク出血熱ウイルスの感染性cDNAの構築
    好井 健太朗, 苅和 宏明, 高島 郁夫, Holbrook Michael
    日本獣医学会学術集会講演要旨集 148回 237 - 237 (公社)日本獣医学会 2009年09月 [査読有り][通常論文]
  • メキシコの野生げっ歯類が保有するハンタウイルスの遺伝子解析
    吉田 喜香, 苅和 宏明, 高野 絢子, 戸谷 理詩, 宮下 大輔, Ngonda Saasa, 瀬戸 隆弘, 真田 崇弘, 吉川 佳佑, 好井 健太朗, 吉松 組子, 有川 次郎, 高島 郁夫
    日本獣医学会学術集会講演要旨集 148回 237 - 237 (公社)日本獣医学会 2009年09月 [査読有り][通常論文]
  • 極東ロシアの野鼠からのハンタウイルスの分離と人におけるウイルス感染状況の調査
    吉川 佳佑, 苅和 宏明, 瀬戸 隆弘, 真田 崇弘, 石塚 万里子, 吉松 組子, 有川 二郎, Ivanov Leonid, Slonova Raisa, 好井 健太朗, 高島 郁夫
    北海道獣医師会雑誌 53 8 112 - 112 (公社)北海道獣医師会 2009年08月 [査読有り][通常論文]
  • 極東ロシアのげっ歯類からのハンタウイルスの分離とウイルス遺伝子の性状解析
    吉川 佳佑, 苅和 宏明, 瀬戸 隆弘, 真田 崇弘, 石塚 万里子, 吉松 組子, 有川 二郎, Ivanov Lenid, 好井 健太朗, 高島 郁夫
    日本獣医学会学術集会講演要旨集 147回 260 - 260 (公社)日本獣医学会 2009年03月 [査読有り][通常論文]
  • 多種類のハンタウイルス血清型の検出が可能な抗原検出法の開発
    真田 崇弘, 苅和 宏明, 瀬戸 隆弘, 谷川 洋一, 宮下 大輔, 吉松 組子, 有川 二郎, 好井 健太朗, 高島 郁夫
    日本獣医学会学術集会講演要旨集 146回 225 - 225 (公社)日本獣医学会 2008年09月 [査読有り][通常論文]
  • ウイルス様粒子を用いたダニ媒介性脳炎の新たな診断法の開発
    千葉 裕美子, 伊川 綾恵, 好井 健太朗, 大森 優紀, 村田 亮, 苅和 宏明, 高島 郁夫
    日本獣医学会学術集会講演要旨集 146回 226 - 226 (公社)日本獣医学会 2008年09月 [査読有り][通常論文]
  • ダニ媒介性脳炎ウイルスの中空ウイルス様粒子のワクチンへの応用
    大森 優紀, 伊川 綾恵, 川上 和江, 好井 健太朗, 苅和 宏明, 高島 郁夫
    日本獣医学会学術集会講演要旨集 146回 226 - 226 (公社)日本獣医学会 2008年09月 [査読有り][通常論文]
  • SARSコロナウイルスのNおよびM蛋白質の粒子形成における機能解析
    中内 美名, 藤井 寛子, 苅和 宏明, 前田 秋彦, 好井 健太朗, 高島 郁夫
    日本獣医学会学術集会講演要旨集 145回 204 - 204 (公社)日本獣医学会 2008年03月 [査読有り][通常論文]
  • ウエストナイルウイルスのE蛋白上糖鎖が宿主内におけるウイルス増殖に与える影響
    村田 亮, 好井 健太朗, 苅和 宏明, 江下 優樹, 高島 郁夫
    日本獣医学会学術集会講演要旨集 145回 205 - 205 (公社)日本獣医学会 2008年03月 [査読有り][通常論文]
  • Arch Virol
    Nakamura, I, Yoshimatsu, K, Lee, B. H, Okumura, M, Taruishi, M, Araki, K, Kariwa, H, Takashima, I, Arikawa, J
    Development of a serotyping ELISA system for Thailand virus infection. 153 1537 1542  2008年 [査読無し][通常論文]
  • ウエストナイルウイルスのE蛋白上糖鎖付加がウイルス増殖に与える影響
    村田 亮, 好井 健太朗, 原田 祐里, 苅和 宏明, 高島 郁夫
    日本獣医学会学術集会講演要旨集 144回 127 - 127 (公社)日本獣医学会 2007年08月 [査読有り][通常論文]
  • ダニ媒介性脳炎ウイルスのウイルス様粒子およびその組換え発現プラスミドを抗原とした針無加圧式注射によるワクチン接種の有効性の検討
    大森 優紀, 伊川 綾恵, 川上 和江, 好井 健太朗, 苅和 宏明, 高島 郁夫
    日本獣医学会学術集会講演要旨集 144回 128 - 128 (公社)日本獣医学会 2007年08月 [査読有り][通常論文]
  • レプリコンを利用したウエストナイルウイルスとダニ媒介性脳炎ウイルスのキメラウイルス様粒子の作製
    原田 祐里, 好井 健太朗, 伊川 綾恵, 苅和 宏明, 高島 郁夫
    日本獣医学会学術集会講演要旨集 142回 121 - 121 (公社)日本獣医学会 2006年08月 [査読有り][通常論文]
  • 苅和宏明, 谷川洋一, 萩谷友洋, LOKUGAMAGE Kumari, LOKUGAMAGE Nandadeva, DAUD Nur Hardy Bin Abu, 好井健太朗, 高島郁夫
    獣医畜産新報 1021 633-638  2006年08月01日 [査読無し][通常論文]
  • ウイルス様粒子を用いたフラビウイルスの粒子形成機構の解析,および診断法・予防法開発への応用
    好井 健太朗, 後藤 明子, 小原 真弓, 川上 和江, 伊川 綾江, 苅和 宏明, 高島 郁夫
    日本獣医学会学術集会講演要旨集 141回 73 - 73 (公社)日本獣医学会 2006年03月 [査読有り][通常論文]
  • ダニ媒介性脳炎ウイルス組み換え蛋白を用いたELISA法による野鼠血清スクリーニング法の開発
    伊川 綾恵, 好井 健太朗, 川上 和江, 後藤 明子, 小原 真弓, 苅和 宏明, 高島 郁夫
    日本獣医学会学術集会講演要旨集 140回 147 - 147 (公社)日本獣医学会 2005年08月 [査読有り][通常論文]
  • 高島 郁夫, 早坂 大輔, 後藤 明子, 好井 健太朗, 苅和 宏明
    ウイルス 55 1 35 - 44 日本ウイルス学会 2005年06月 [査読有り][通常論文]
     
    北海道と極東ロシアのダニ媒介性脳炎ウイルスの系統解析の結果,北海道株は極東地区において数百年前に出現したと推定された.イルクーツク地区のダニ媒介性脳炎ウイルスはシベリア亜型と同定された.ダニ媒介性脳炎ウイルスのBHK細胞適応変異株はマウスにおける神経侵襲性毒力が低下していた.変異株はエンベロープ蛋白に1ヶ所のアミノ酸置換があり,荷電が陽性に変化する変異であった.変異株はウイルス血症と脾臓でのウイルス価が親株に比べ低下していた.ダニ媒介性脳炎ウイルスの感染性cDNAクローンの作製に成功し,神経毒力の解析を行った.エンベロープ蛋白の1ヶ所とNs5の2ヶ所のアミノ酸変異が相乗的に神経毒力の低下に関与していた(著者抄録)
  • 有川二郎, 吉松組子, KUMPERASART Sanit, THANG Truong Thua, 荒木幸一, LEE Byoung‐Hee, KRUGER Detlev H, 苅和宏明, 高島郁夫
    獣医畜産新報 1005 326 - 328 2005年04月01日 [査読無し][通常論文]
  • 野生げっ歯類からのハンタウイルス抗原検出用ELISAの開発
    谷川 洋一, 苅和 宏明, 萩谷 友洋, Lokugamage Nandadeva, Lokugamage Kumari, 舘 敦史, 好井 健太朗, 吉松 組子, 有川 二郎, 高島 郁夫
    日本獣医学会学術集会講演要旨集 139回 185 - 185 (公社)日本獣医学会 2005年03月 [査読有り][通常論文]
  • フラビウイルスのウイルス粒子分泌におけるユビキチン-プロテアソーム系の関与
    好井 健太朗, 後藤 明子, 川上 和江, 苅和 宏明, 高島 郁夫
    日本獣医学会学術集会講演要旨集 138回 129 - 129 (公社)日本獣医学会 2004年08月 [査読有り][通常論文]
  • 日本におけるハンタウイルス感染の動物伝染病学的及び疫学的研究(Epizootiological and epidemiological study of hantavirus infection in Japan)
    Lokugamage Nandadeva, 苅和 宏明, 好井 健太朗, 舘 敦史, 安藤 秀二, 福島 博, 土屋 公幸, 岩崎 琢也, 吉松 組子, 有川 二郎, 高島 郁夫
    日本獣医学会学術集会講演要旨集 138回 130 - 130 2004年08月 [査読有り][通常論文]
  • タイリクヤチネズミから分離されたPuumala型近縁ハンタウイルスの遺伝子解析
    谷川 洋一, 苅和 宏明, Lokugamage Nandadeva, 萩谷 友洋, Lokugamage Kumari, 舘 敦史, 好井 健太朗, 岩佐 真宏, 高島 郁夫
    日本獣医学会学術集会講演要旨集 138回 130 - 130 (公社)日本獣医学会 2004年08月 [査読有り][通常論文]
  • Amur型ハンタウイルス糖蛋白の哺乳類細胞での発現と抗原性解析
    舘 敦史, 苅和 宏明, Lokugamage Kumari, Lokugamage Nandadeva, 谷川 洋一, 好井 健太朗, 吉松 組子, 有川 二郎, 高島 郁夫
    日本獣医学会学術集会講演要旨集 138回 131 - 131 (公社)日本獣医学会 2004年08月 [査読有り][通常論文]
  • ダニ媒介性脳炎ウイルス組み替え蛋白を用いたELISAによる野ネズミ血清スクリーニング法の開発
    川上 和江, 好井 健太朗, 後藤 明子, 苅和 宏明, 高島 郁夫
    日本獣医学会学術集会講演要旨集 137回 114 - 114 (公社)日本獣医学会 2004年03月 [査読有り][通常論文]
  • repliconを利用したフラビウイルスのキメラウイルス様粒子の作成
    好井 健太朗, 早坂 大輔, 後藤 明子, 苅和 宏明, 高島 郁夫
    日本獣医学会学術集会講演要旨集 137回 114 - 114 (公社)日本獣医学会 2004年03月 [査読有り][通常論文]
  • ダニ媒介性脳炎ウイルスのE蛋白の糖鎖修飾がウイルス粒子分泌に与える影響
    後藤 明子, 好井 健太朗, 小原 真弓, 植木 智隆, 水谷 哲也, 苅和 宏明, 高島 郁夫
    日本獣医学会学術集会講演要旨集 137回 115 - 115 (公社)日本獣医学会 2004年03月 [査読有り][通常論文]
  • ダニ媒介性脳炎ウイルスの感染性cDNAクローンを用いた病原性解析
    植木 智隆, 早坂 大輔, 後藤 明子, 好井 健太朗, 水谷 哲也, 苅和 宏明, 高島 郁夫
    日本獣医学会学術集会講演要旨集 136回 176 - 176 (公社)日本獣医学会 2003年09月 [査読有り][通常論文]
  • ダニ媒介性脳炎ウイルスのrepliconおよびpackaging systemの構築
    好井 健太朗, 早坂 大輔, 後藤 明子, 水谷 哲也, 苅和 宏明, 高島 郁夫
    日本獣医学会学術集会講演要旨集 136回 257 - 257 (公社)日本獣医学会 2003年09月 [査読有り][通常論文]
  • ダニ媒介性脳炎ウイルスのエンベロープ糖蛋白の糖鎖欠損がウイルス粒子形成に与える影響
    後藤 明子, 好井 健太朗, 小原 真弓, 植木 智隆, 水谷 哲也, 苅和 宏明, 高島 郁夫
    日本獣医学会学術集会講演要旨集 135回 156 - 156 (公社)日本獣医学会 2003年03月 [査読有り][通常論文]
  • ダニ媒介性脳炎ウイルスのウイルス粒子放出抑制に関する解析
    好井 健太朗, 後藤 明子, 小原 真弓, 植木 智隆, 早坂 大輔, 水谷 哲也, 苅和 宏明, 高島 郁夫
    日本獣医学会学術集会講演要旨集 134回 195 - 195 (公社)日本獣医学会 2002年08月 [査読有り][通常論文]
  • ダニ媒介性脳炎ウイルス神経侵入性弱毒変異株のマウス体内での動態
    後藤 明子, 好井 健太朗, 早坂 大輔, 水谷 哲也, 苅和 宏明, 高島 郁夫
    日本獣医学会学術集会講演要旨集 134回 195 - 195 (公社)日本獣医学会 2002年08月 [査読有り][通常論文]
  • 北海道におけるダニ媒介性脳炎ウイルスの血清疫学調査
    小原 真弓, 好井 健太朗, 後藤 明子, 早坂 大輔, 水谷 哲也, 苅和 宏明, 高島 郁夫
    日本獣医学会学術集会講演要旨集 134回 195 - 195 (公社)日本獣医学会 2002年08月 [査読有り][通常論文]
  • Daisuke Hayasaka, Akiko Goto, Kentaro Yoshii, Tetsuya Mizutani, Hiroaki Kariwa, Ikuo Takashima
    Vaccine 19 32 4774 - 4779 2001年09月 [査読無し][通常論文]
  • Araki K, Yoshimatsu K, Ogino M, Ebihara H, Lundkvist A, Kariwa H, Takashima I, Arikawa J
    Journal of clinical microbiology 39 2397 - 2404 7 2001年07月 [査読有り][通常論文]
  • Arikawa J, Yoshimatsu K, Kariwa H
    Japanese journal of infectious diseases 54 95 - 102 3 2001年06月 [査読有り][通常論文]
  • Distribution and characterization of tick-borne encephalitis viruses in Siberia and far-eastern region
    Daisuke Hayasaka, Leonid Ivanov, Galina N. Leonova, Akiko Goto, Kentaro Yoshii, Tetsuya Mizutani, Hiroaki Kariwa, Ikuo Takashima
    J. Gen. Virol. 82 Pt6 1319 - 1328 2001年06月 [査読無し][通常論文]
  • Michael E. Murphy, Hiroaki Kariwa, Tetsuya Mizutani, Hiroki Tanabe, Kumiko Yoshimatsu, Jiro Arikawa, Ikuo Takashima
    J Vet Med Sci. 63 6 637 - 645 2001年06月 [査読無し][通常論文]
  • ダニ媒介性脳炎(TBE)ウイルスシベリア型及び極東型の病原性の比較とワクチンの効果
    早坂 大輔, 後藤 明子, 好井 健太朗, 水谷 哲也, 苅和 宏明, 高島 郁夫
    日本獣医学会学術集会講演要旨集 131回 121 - 121 (公社)日本獣医学会 2001年03月 [査読有り][通常論文]
  • ダニ媒介性脳炎ウイルス組み換え蛋白の作成と抗原性状の解析
    好井 健太朗, 早坂 大輔, 後藤 明子, 水谷 哲也, 苅和 宏明, 高島 郁夫
    日本獣医学会学術集会講演要旨集 131回 121 - 121 (公社)日本獣医学会 2001年03月 [査読有り][通常論文]
  • Epidemiology of tick-borne encephalitis (TBE) and phylogenetic analysis of TBE viruses in Japan and Far Eastern Russia
    Ikuo Takashima, Daisuke Hayasaka, Akiko Goto, Hiroaki Kariwa, Tetsuya Mizutani
    Jpn J Infect Dis. 54 1 1 - 11 2001年01月 [査読無し][通常論文]
  • In vitro antiviral activity of lactoferrin and ribavirin upon hantavirus
    Michael E. Murphy, Hiroaki Kariwa, Tetsuya Mizutani, Kumiko Yoshimatsu, Jiro Arikawa, Ikuo Takashima
    Arch. Virol. 145 8 1571 - 1582 2000年08月 [査読無し][通常論文]
  • Tetsuya Mizutani, Hisae Inagaki, Mitsuhiro Tada, Daisuke Hayasaka, Michael E. Murphy, Toshiyoshi Fujiwara, Jun-ichi Hamada, Hiroaki Kariwa, Ikuo Takashima
    Microbirol. Immunol. 44 7 597 - 603 2000年07月 [査読無し][通常論文]
  • Characterization of monoclonal antibodies against Hokkaido strain tick-borne encephalitis virus
    Kimiyo Komoro, Daisuke Hayasaka, Tetsuya Mizutani, Hiroaki Kariwa, Ikuo Takashima
    Microbiol Immunol. 44 6 533 - 536 2000年06月 [査読無し][通常論文]
  • Hiroaki Kariwa, Kumiko Yoshimatsu, Kouichi Araki, Chayama K, Kumada H, Michiko Ogino, Ebihara H, Michael E. Murphy, Tetsuya Mizutani, Ikuo Takashima, Jiro Arikawa
    Microbiol Immunol. 44 5 357 - 362 2000年05月 [査読無し][通常論文]
  • Phylogenetic and virulence analysis of Tick-borne encephalitis viruses from Japan and Far East Russia
    Daisuke Hayasaka, Yoshiyuki Suzuki, Hiroaki Kariwa, Leonid Ivanov, Vladimir Volkov, Vladimir Demenev, Tetsuya Mizutani, Takashi Gojobori, Ikuo Takashima
    J. Gen. Virol. 80 Pt12 3127 - 3135 1999年12月 [査読無し][通常論文]
  • Tetsuya Mizutani, Hisae Inagaki, Daisuke Hayasaka, Hiroaki Kariwa, Ikuo Takashima
    J. Vet. Med. Sci. 61 7 831 - 834 1999年07月 [査読無し][通常論文]
  • 腎症候性出血熱
    苅和宏明, 水谷哲也, 高島郁夫
    「最新医学」 6月増刊号 54 1425 - 1431 1999年06月 [査読無し][通常論文]
  • Nobuyuki Chiba, Mihoro Osada, Kimiyo Komoro, Tetsuya Mizutani, Hiroaki Kariwa, Ikuo Takashima
    Vaccine 17 11-12 1532 - 1539 1999年03月 [査読無し][通常論文]
  • T Mizutani, M Ogino, Y Nishino, T Kimura, H Inagaki, D Hayasaka, H Kariwa, Takashima, I
    JAPANESE JOURNAL OF VETERINARY RESEARCH 46 4 165 - 169 1999年02月 [査読有り][通常論文]
     
    There are few copies of Borna disease virus (BDV) genome in peripheral blood mononuclear cells and no reliable standard reverse transcriptase-polymerase chain reaction (RT-PCR) method for the detection of BDV RNA, which is both highly sensitive and free of contamination. Single-step RT-PCR, in which both reverse transcription and amplification by Tag DNA polymerase work efficiently in a single buffer, was applied to detect the p24 region of BDV RNA in vitro and in vivo. Using in vitro synthesized RNA, it was demonstrated that at least 100 copies of BDV RNA could be detected and the sensitivity and specificity were nearly equal to those obtained by RT-nested PCR. We could detect BDV RNA from more than 1 pg of cellular RNA obtained from BDV-persistently infected MDCK cells. Furthermore, this method was successfully performed on brain specimens obtained from a BDV-infected rat at II weeks post-inoculation. This single-step RT-PCR method will be convenient for detecting limited amounts of BDV RNA in various cells and tissue samples.
  • Nobuyuki Chiba, Takuya Iwasaki, Tetsuya Mizutani, Hiroaki Kariwa, Takeshi Kurata, Ikuo Takashima
    Vaccine 17 7-8 779 - 787 1999年02月 [査読無し][通常論文]
  • Koji Tsujimura, Tetsuya Mizutani, Kumiko Yoshimatsu, Michiko Oginio, Yuko Morii, Hisae Inagaki, Jiro Arikawa, Hiroaki Kariwa, Ikuo Takashima
    J. Vet. Med. Sci. 61 2 113 - 117 1999年02月 [査読無し][通常論文]
  • T Mizutani, H Inagaki, D Hayasaka, S Shuto, N Minakawa, A Matsuda, H Kariwa, Takashima, I
    ARCHIVES OF VIROLOGY 144 10 1937 - 1946 1999年 [査読有り][通常論文]
     
    Regulation of viral RNA levels in infected cells is considered important in the investigation of viral transcription and replication. Amounts of Borna disease virus (BDV) RNAs were increased in confluent persistently BDV-infected MDCK cells (MDCK/BDV) cells, while maintained at low levels in growing cells. The amount of 1.9-kb RNA without cap formation and polyadenylation at the 5' and 3' ends respectively were remarkably increased (200% per day) in confluent MDCK/BDV cells. Both the full-length genomic and anti-genomic RNAs were increased accompained by 1.9-kb RNA, suggesting the transcription of the 1.9-kb RNA was important for replication of BDV. Ribavirin has an inhibitory effect on replication and transcription of BDV at concentrations from 1 to 10 mu g/ml [Mizutani T et al., Arch Virol (1998) 143: 2039-2044]. BDV transcripts were decreased with ribavirin treatment and increased after its removal which indicated that ribavirin has a reversible inhibitory effect on BDV transcription. Furthermore, BDV transcription was also decreased by two agents, RMNPA and EICAR, which selectively inhibit enzyme activity related to cap formation at the 5' end of mRNA. On the contrary, when the growing MDCK/BDV cells were treated with actinomycin D, transcripts of BDV RNA were increased for 24 h. These agents and culture conditions in this study were found to be useful tools for up-and down-regulation of BDV transcription in persistently BDV-infected cells.
  • Tetsuya Mizutani, Yoshii Nishino, Hiroaki Kariwa, Ikuo Takashima
    J. Vet. Med. Sci. 61 1 77 - 80 1999年01月 [査読無し][通常論文]
  • 高島郁夫, 有川二郎, 苅和宏明, 根占哲也
    北海道の研究開発 1998 95 - 100 1998年12月 [査読無し][通常論文]
  • Mizutani T, Ogino M, Nishino Y, Kimura T, Kariwa H, Tsujimura K, Inagaki H, Takashima I
    The Japanese journal of veterinary research 46 73 - 81 2-3 1998年11月 [査読有り][通常論文]
  • Tetsuya Mizutani, Hisae Inagaki, Koichi Araki, Hiroaki Kariwa, Jiro Arikawa, Ikuo Takashima
    Arch. Virol. 143 10 2039 - 2044 1998年10月 [査読無し][通常論文]
  • M Morii, K Yoshimatsu, J Arikawa, GZ Zhou, H Kariwa, Takashima, I
    JOURNAL OF CLINICAL MICROBIOLOGY 36 9 2514 - 2521 1998年09月 [査読有り][通常論文]
     
    Hantaan virus (HTN) and Seoul virus (SEO) are members of the genus Hantavirus in the family Bunyaviridae and are causative agents of hemorrhagic fever with renal syndrome. The complete and truncated nucleocapsid proteins (NP) of HTN and SEO were expressed by a recombinant baculovirus system, Antigenic characterization of the NP using monoclonal antibodies (Mabs)indicated that the binding sites For the serotype-specific MAbs were located between amino acids (aa) 155 and 429, a Western blot assay indicated that the serotype-specific epitopes were conformation dependent. An indirect immunofluorescence antibody (IFA) assay with the truncated NP (aa 155 to 429) was able to distinguish convalescent-phase sera from HTN and SEO patients. However, the antibody liters with the truncated NP were lower than those with the whole NP, The truncated NP of SEO (aa 155 to 429) could be used as an enzyme-linked immunosorbent assay (ELISA) antigen, but the truncated NP from HTN lost its reactivity when used for ELISA. The LFA assay using baculovirus-expressed truncated NP as an antigen is a rapid, simple, and safe test for distinguishing between HTN and SEO infections by serotype.
  • H Kariwa, M Fujiki, K Yoshimatsu, J Arikawa, Takashima, I, N Hashimoto
    ARCHIVES OF VIROLOGY 143 2 365 - 374 1998年 [査読有り][通常論文]
     
    To understand the mode of transmission of Seoul type hantavirus in Wistar rats, we examined the shedding of the virus and antibody production in infected rats. When 1-day-old rats were inoculated with KI-83-262 strain of Seoul virus, the S segment of the viral genome was detected in lungs, clots, urine, saliva, submaxillary glands, rectums, and-kidneys by nested reverse transcriptase PCR. On the other hand, when 8-week-old rats were infected with the virus, viral genome was detected only in the lungs and rectum. In uninfected newborn rats intranasally administered urine from infected newborn rats, four of six rats shed the virus into their urine. In addition, three of eight rats kept in a same cage with infected animals also shed the virus into urine. Moreover, the virus genome was detected in the urine of urban rats (Rattus norvegicus) in an enzootic focus. These findings suggest that the urine containing virus from infected rats is an actual source of Seoul virus infection.
  • Takashima, I, M Hiyoshi, H Kariwa, R Mukaiya, N Hashimoto
    MICROBIOLOGY AND IMMUNOLOGY 40 4 265 - 270 1996年 [査読有り][通常論文]
     
    Japanese quail were used for the infection model of avian chlamydiosis. One-day-old Japanese quail were highly susceptible to lethal infection by a Chlamydia psittaci strain of budgerigar origin upon inoculation via the air sac route with 10(4.1) FFU of the organism, showing an acute and lethal course with chlamydial propagation. In contrast, 7-day-old quail developed resistance to the infection as shown by the lack of lethal effect with the same dose. The resistance of 7-day-old birds was abolished by immunosuppressive treatment with cyclophosphamide. Upon inoculation with a sublethal dose of 10(2.1) FFU, latent infection was established in 1-day-old birds with a minimum number of the organism. The latent infection in the birds was converted to the lethal form by treatment with cyclophosphamide along with chlamydial propagation and suppression of antibody production.
  • Kariwa H, Kimura M, Yoshizumi S, Arikawa J, Yoshimatsu K, Takashima I, Hashimoto N
    Archives of virology 141 12 2327 - 2338 12 1996年 [査読有り][通常論文]
  • Yoshimatsu K, Arikawa J, Yoshida R, Li H, Yoo YC, Kariwa H, Hashimoto N, Kakinuma M, Nobunaga T, Azuma I
    Laboratory animal science 45 641 - 646 6 1995年12月 [査読有り][通常論文]
  • Kariwa H, Yoshizumi S, Arikawa J, Yoshimatsu K, Takahashi K, Takashima I, Hashimoto N
    The American journal of tropical medicine and hygiene 53 222 - 227 3 1995年09月 [査読有り][通常論文]
  • Kariwa H, Kamimura M, Arikawa J, Yoshimatsu K, Takashima I, Hashimoto N
    Microbiology and immunology 39 35 - 41 1 1995年 [査読有り][通常論文]
  • Kariwa H, Arikawa J, Takashima I, Isegawa Y, Yamanishi K, Hashimoto N
    Journal of virological methods 49 2 235 - 244 2 1994年09月 [査読有り][通常論文]
  • Kariwa H, Isegawa Y, Arikawa J, Takashima I, Ueda S, Yamanishi K, Hashimoto N
    Virus research 33 1 27 - 38 1 1994年07月 [査読有り][通常論文]
  • JG GARCIA, TAKASHIMA, I, H KARIWA, N HASHIMOTO
    JOURNAL OF VIROLOGICAL METHODS 48 1 31 - 41 1994年06月 [査読有り][通常論文]
     
    A modified antibody-forming cell assay was used to enumerate splenocytes secreting antibodies to Japanese encephalitis (JE) virus and four other flaviviruses in infected cells as the target antigens. The optimal viral antigen expression for the assay was standardized in the infected cells for each virus and the incubation time varied depending of the cell line and the virus. The kinetics of response of JE virus-infected mice readily showed IgG isotype switching. Antibody-forming splenocytes of mice primed or boosted with one flavivirus could distinguish, in variable proportions, the homologous virus antigen from heterologous ones depending on the serocomplex of the virus. Antibody-forming cells from flavivirus-infected mice were flavivirus specific as evidenced by the lack of recognition of Getah virus (alphavirus) antigen. After JE virus-infected mice were cross-primed with another flavivirus, the IgG-forming cell response resembled the secondary response to both viruses but had higher affinity to JE virus than to the cross-priming virus.
  • Arikawa J, Ito M, Yao JS, Kariwa H, Takashima I, Hashimoto N
    The Journal of veterinary medical science / the Japanese Society of Veterinary Science 56 1 27 - 32 1 1994年02月 [査読有り][通常論文]
  • Yoshimatsu K, Arikawa J, Kariwa H
    The Journal of veterinary medical science / the Japanese Society of Veterinary Science 55 6 1047 - 1050 6 1993年12月 [査読有り][通常論文]
  • J GARCIAGARCIA, TAKASHIMA, I, H KARIWA, N HASHIMOTO
    JOURNAL OF IMMUNOLOGICAL METHODS 157 1-2 259 - 267 1993年01月 [査読有り][通常論文]
     
    A sensitive modified method for the detection in vitro of antibody-forming cells (AFCs) in Japanese encephalitis (JE) virus infected mice is described. The procedure involves the selection of the most appropriate incubation period, after the infection of monolayer cells, for expression of viral antigens to be recognized by AFCs. Adequate preservation of the expressed antigens was achieved after fixation with paraformaldehyde-lysine-periodate buffer (PLP). The development of spots which corresponded to AFCs was successfully obtained with the use of the avidin-biotin complex (ABC) amplification reaction. The number of the spots developing after completion of the assay was greater after PLP fixation compared with other fixation solutions such as methanol, formalin and acetone-water. Optimum antigen expression on the infected cells for recognition by lymphoid cells was reached after 24 h of infection. However, it was possible to detect viral antigens on the infected cells 8 h after infection. The ABC reaction was shown to be the best procedure for developing spots compared with other immunocytochemical methods. A linear increment was observed in the number of spots that developed at different spleen cell densities. This assay is simple to perform and could detect virus-specific Ig producing cells from the spleens of mice infected with JE virus. It also makes it possible to enumerate accurately the kinetics of the AFC response in different lymphoid organs during infection or after vaccination.
  • JS YAO, J ARIKAWA, H KARIWA, K YOSHIMATSU, TAKASHIMA, I, N HASHIMOTO
    JAPANESE JOURNAL OF VETERINARY RESEARCH 40 2-3 87 - 97 1992年09月 [査読有り][通常論文]
     
    The effect of neutralizing monoclonal antibodies (N MAbs) on Hantaan virus infection of macrophages was investigated using P388D1 cells, a murine macrophage cell line. MAbs to the G1 protein (G1b) and the G2 protein (G2a and G2c) neutralized viral infectivity in P388D1 cells. N MAbs to G1b showed much higher neutralizing potency than those to G2a and G2c. With each N MAbs, two distinct effects were observed: neutralization of viral infectivity occurring at high concentrations and enhancement of that at low concentrations. Non-neutralizing MAbs, on the other hand, showed only enhancement of viral infectivity even at high concentrations without any inhibitory effects. The Fab fragments of N MAbs showed neither neutralizing nor enhancing activities. However, when the virus-Fab complexes were reacted with the anti-Fab antibodies, both neutralization and enhancement of viral infectivity were restored depending on the dose of Fab fragments. These results indicate that Hantaan virus infection of P388D1 cells is mediated by the Fc portion of the antibodies and neutralization is dependent on the concentration of N antibodies bound bivalently to the neutralization site on the virion.
  • H KARIWA, J ARIKAWA, TAKASHIMA, I, N HASHIMOTO
    JOURNAL OF VIROLOGICAL METHODS 37 3 345 - 354 1992年06月 [査読有り][通常論文]
     
    A new serodiagnostic method designated protein-G antibody assay (PGA) was developed for detection of hantavirus infection in various species of animals. The assay procedure includes reacting the sera with hantavirus-infected cells on glass slides, followed by incubation of biotinylated protein G and amplification with the avidin-biotinylated peroxidase complex. Specific antibody in rabbit, rat, mouse and Mongolian gerbil serum was detected by this method. The PGA titres were similar to those of the neutralization titre. In the sera of Mongolian gerbils infected with strain SR-11, antibody was first detected 10 days post infection, and the titre increased to 1:256 at 18 days post-infection. PGA was evaluated using sera of urban rats (Rattus norvegicus) captured in an endemic area of hantavirus infection. The negative (less-than-or-equal-to 1:1, 24/62, 38.7%) and positive groups (greater-than-or-equal-to 1:16, 38/62, 61.3%) were clearly distinguished. PGA titres were closely related to IFA titres in the sera. Two of 10 sera from Clethrionomys rufocanus and one from Apodemus speciosus captured in the same endemic area were positive to both PGA and IFA. These data indicate that PGA is a simple and useful method for seroepizootiological surveys of hantavirus infection, especially in wild rodent reservoirs.
  • JS YAO, H KARIWA, TAKASHIMA, I, K YOSHIMATSU, J ARIKAWA, N HASHIMOTO
    ARCHIVES OF VIROLOGY 122 1-2 107 - 118 1992年 [査読有り][通常論文]
     
    Antibody-dependent enhancement (ADE) of hantavirus infections (strains Hantaan 76-118 and SR-11) was studied using macrophage-like cell lines (J774.1, P388D 1, and U937). Significantly higher virus titers (1,000 to 4,000 FFU/ml) were obtained by pretreatment of the virus with immune serum as compared to normal serum (< 20 FFU/ml). Monoclonal antibodies (MAbs) to strain Hantaan 76-118 were employed to determine the antigenic determinants responsible for the ADE activity. ADE of the infection occurred with MAbs to both G1 and G2 envelope glycoproteins, but not with MAbs to nucleocapsid protein. Antigenic determinants related to haemagglutination or virus neutralization were found to cause ADE of the infection.
  • 苅和宏明, 有川二郎, よう健勝, 高島郁夫, 橋本信夫
    北海道獣医師会雑誌 35 3 69 - 74 1991年03月 [査読無し][通常論文]

MISC

  • ウエストナイルウイルスのカプシドタンパク質とAMPKの相互作用の解析
    鈴木 健矢, 小林 進太郎, 好井 健太朗, 苅和 宏明 日本獣医学会学術集会講演要旨集 162回 420 -420 2019年08月 [査読無し][通常論文]
  • 2017、2018年に北海道道央地域のヤマトマダニから分離されたダニ媒介性脳炎ウイルスの性状解析
    高橋 侑嗣, 小林 進太郎, 石塚 万里子, 中尾 亮, 苅和 宏明, 好井 健太朗 日本獣医学会学術集会講演要旨集 162回 421 -421 2019年08月 [査読無し][通常論文]
  • ダニ媒介性脳炎ウイルス由来非コードRNAと相互作用するマダニ宿主蛋白質の同定及び機能解析
    西山 祥子, 平野 港, 武藤 芽未, 小林 進太郎, 神谷 亘, 苅和 宏明, 好井 健太朗 日本獣医学会学術集会講演要旨集 162回 421 -421 2019年08月 [査読無し][通常論文]
  • エンベロープタンパク質のアミノ酸変異によるウエストナイルウイルスの増殖および病態形成への影響
    小林 進太郎, 金子 知里, 川上 怜子, 長谷部 理絵, 澤 洋文, 好井 健太朗, 苅和 宏明 日本獣医学会学術集会講演要旨集 162回 431 -431 2019年08月 [査読無し][通常論文]
  • 中釜尚人, 平野港, 武藤芽未, 西山祥子, 中尾亮, 小林進太郎, 苅和宏明, 好井健太朗 日本獣医学会学術集会講演要旨集 161st 383 2018年08月21日 [査読無し][通常論文]
  • ダニ媒介性脳炎ウイルスのウイルス様粒子を用いた新規IgM-ELISA系の開発
    中安 美樹, 小林 進太郎, 平野 港, 武藤 芽未, 苅和 宏明, 好井 健太朗 日本獣医学会学術集会講演要旨集 161回 384 -384 2018年08月 [査読無し][通常論文]
  • 2017年に北海道道央地域で分離されたダニ媒介性脳炎ウイルスの性状解析
    高橋 侑嗣, 石塚 万里子, 平野 港, 武藤 芽未, 西山 祥子, 小林 進太郎, 苅和 宏明, 好井 健太朗 日本獣医学会学術集会講演要旨集 161回 384 -384 2018年08月 [査読無し][通常論文]
  • ダニ媒介性脳炎ウイルス由来非コードRNAの感染時における機能解析
    西山 祥子, 平野 港, 武藤 芽未, 神原 真生, 小林 進太郎, 苅和 宏明, 好井 健太朗 日本獣医学会学術集会講演要旨集 161回 384 -384 2018年08月 [査読無し][通常論文]
  • ユビキチンの蓄積に着目したウエストナイルウイルスの病原性解析
    川上 怜子, 小林 進太郎, 好井 健太朗, 苅和 宏明 日本獣医学会学術集会講演要旨集 161回 384 -384 2018年08月 [査読無し][通常論文]
  • Hokkaidoウイルスのエンベロープ糖タンパク質Gnの組換えタンパク質発現とその応用
    岡崎 はるか, 小林 進太郎, 好井 健太朗, 苅和 宏明 日本獣医学会学術集会講演要旨集 161回 385 -385 2018年08月 [査読無し][通常論文]
  • 北海道のアライグマを対象としたダニ媒介性脳炎ウイルスの血清疫学調査
    佐鹿 万里子, 石塚 万里子, 三瓶 孝男, 小林 進太郎, 西山 祥子, 苅和 宏明, 好井 健太朗 日本獣医学会学術集会講演要旨集 161回 394 -394 2018年08月 [査読無し][通常論文]
  • 北海道と福島県の野生動物におけるダニ媒介性脳炎ウイルス(TBEV)の血清学的調査(A serological survey for tick-borne encephalitis virus(TBEV) in wild animals in Hokkaido and Fukushima prefecture)
    Dulamjav Jamsransuren, 小川 晴子, 佐々木 基樹, 福本 晋也, 松本 高太郎, 好井 健太朗, 苅和 宏明, 浅川 満彦, 奥田 圭, 山口 英美 日本獣医学会学術集会講演要旨集 161回 395 -395 2018年08月 [査読無し][通常論文]
  • ウエストナイルウイルス感染で起こるオートファジーの抑制と神経病態形成の関係
    小林 進太郎, 好井 健太朗, Wallaya Phongpaew, 武藤 芽未, 平野 港, 大場 靖子, 澤 洋文, 苅和 宏明 日本獣医学会学術集会講演要旨集 161回 395 -395 2018年08月 [査読無し][通常論文]
  • ウエストナイルウイルスのカプシドタンパク質によるオートファジーの抑制による変性タンパク質の蓄積と神経病態形成についての解析
    小林 進太郎, 好井 健太朗, Phonqphaew Wallaya, 武藤 芽未, 平野 港, 大場 靖子, 澤 洋文, 苅和 宏明 生命科学系学会合同年次大会 2017年度 [3P -1110] 2017年12月 [査読無し][通常論文]
  • ウエストナイルウイルスの脳内侵入におけるエンベロープタンパク質のアミノ酸の解析
    金子 知里, 小林 進太郎, 好井 健太朗, 苅和 宏明 日本獣医学会学術集会講演要旨集 160回 411 -411 2017年08月 [査読無し][通常論文]
  • ダニ媒介性ウイルス感染におけるダニRNAi関連因子の機能解析
    神原 真生, 平野 港, 武藤 芽未, 石塚 万里子, 小林 進太郎, 苅和 宏明, 好井 健太朗 日本獣医学会学術集会講演要旨集 160回 411 -411 2017年08月 [査読無し][通常論文]
  • ダニ媒介性脳炎ウイルスの3'非翻訳領域に関わる宿主因子の検索
    武藤 芽未, 神谷 亘, 境 瑞紀, 平野 港, 小林 進太郎, 苅和 宏明, 好井 健太朗 日本獣医学会学術集会講演要旨集 160回 411 -411 2017年08月 [査読無し][通常論文]
  • Hokkaidoウイルスと他のハンタウイルスの共感染による増殖性の補完に関する研究
    梶河 紗代, 岩崎 里奈, 真田 崇弘, 小林 進太郎, 好井 健太朗, 苅和 宏明 日本獣医学会学術集会講演要旨集 159回 408 -408 2016年08月 [査読無し][通常論文]
  • フラビウイルスゲノムの神経細胞内輸送機構の解析
    平野 港, 境 瑞紀, 武藤 芽未, 小林 進太郎, 苅和 宏明, 好井 健太朗 日本獣医学会学術集会講演要旨集 159回 409 -409 2016年08月 [査読無し][通常論文]
  • ウエストナイルウイルスの粒子放出過程におけるRab8bタンパク質の役割
    小林 進太郎, 鈴木 忠樹, 川口 晶, Phongphaew Wallaya, 好井 健太朗, 苅和 宏明, 大場 靖子, 澤 洋文 日本獣医学会学術集会講演要旨集 159回 409 -409 2016年08月 [査読無し][通常論文]
  • In trans補完系によるダニ媒介性脳炎ウイルスのゲノム複製機構の解析
    山内 沙也果, 小林 進太郎, 平野 港, 武藤 芽未, 石塚 万里子, 苅和 宏明, 好井 健太朗 日本獣医学会学術集会講演要旨集 159回 409 -409 2016年08月 [査読無し][通常論文]
  • ダニ媒介性脳炎ウイルスのNucleoside Inhibitor耐性変異の解析
    近藤 寛史, 平野 港, 石塚 万里子, 武藤 芽未, 小林 進太郎, 苅和 宏明, Daniel Ruzek, 好井 健太朗 日本獣医学会学術集会講演要旨集 159回 423 -423 2016年08月 [査読無し][通常論文]
  • ダニ媒介性脳炎ウイルスの神経細胞内におけるウイルスゲノムRNA輸送機構の解析
    平野 港, 境 瑞紀, 苅和 宏明, 小林 進太郎, 好井 健太朗 日本生化学会大会・日本分子生物学会年会合同大会講演要旨集 88回・38回 [1P1134] -[1P1134] 2015年12月 [査読無し][通常論文]
  • ウエストナイルウイルス感染による変性タンパク質蓄積機構の解析
    小林 進太郎, Phongphaew Wallaya, 好井 健太朗, 平野 港, 武藤 芽未, 大場 靖子, 澤 洋文, 苅和 宏明 日本生化学会大会・日本分子生物学会年会合同大会講演要旨集 88回・38回 [1P1135] -[1P1135] 2015年12月 [査読無し][通常論文]
  • ダニ媒介性脳炎ウイルスのウイルス様粒子を用いた動物種非特異的な新規血清学的診断法の開発
    稲垣 恵理, 境 瑞紀, 平野 港, 武藤 芽未, 苅和 宏明, 好井 健太朗 日本獣医学会学術集会講演要旨集 158回 365 -365 2015年08月 [査読無し][通常論文]
  • イムノクロマトグラフィー法によるハンタウイルスの迅速診断法の開発
    小山 芽以, 吉松 組子, 好井 健太朗, 有川 二郎, 苅和 宏明 日本獣医学会学術集会講演要旨集 158回 373 -373 2015年08月 [査読無し][通常論文]
  • 中尾亮, 梶原将大, 邱永晋, 森亜紀奈, 直亨則, 村松美笑子, 好井健太朗, 苅和宏明, 澤洋文, 杉本千尋, 高田礼人 日本獣医学会学術集会講演要旨集 157th 451 2014年08月11日 [査読無し][通常論文]
  • モンゴルにおけるダニ媒介性脳炎ウイルスの分離と性状解析
    武藤 芽未, Bazartseren Boldbaatar, 好井 健太朗, 苅和 宏明 日本獣医学会学術集会講演要旨集 157回 439 -439 2014年08月 [査読無し][通常論文]
  • レポーター遺伝子発現ダニ媒介性脳炎ウイルスの作製と性状解析
    池端 真帆, 好井 健太朗, 境 瑞紀, 平野 港, 苅和 宏明 日本獣医学会学術集会講演要旨集 157回 450 -450 2014年08月 [査読無し][通常論文]
  • 苅和宏明, 好井健太朗, 高島郁夫 日本臨床 618 -621 2013年12月20日 [査読無し][通常論文]
  • Hiroaki Kariwa, Ryo Murata, Masashi Totani, Kentaro Yoshii, Ikuo Takashima INTERNATIONAL JOURNAL OF ENVIRONMENTAL RESEARCH AND PUBLIC HEALTH 10 (12) 7144 -7164 2013年12月 [査読無し][通常論文]
     
    In this review, we discuss the possibility that the glycosylation of West Nile (WN) virus E-protein may be associated with enhanced pathogenicity and higher replication of WN virus. The results indicate that E-protein glycosylation allows the virus to multiply in a heat-stable manner and therefore, has a critical role in enhanced viremic levels and virulence of WN virus in young-chick infection model. The effect of the glycosylation of the E protein on the pathogenicity of WN virus in young chicks was further investigated. The results indicate that glycosylation of the WN virus E protein is important for viral multiplication in peripheral organs and that it is associated with the strong pathogenicity of WN virus in birds. The micro-focus reduction neutralization test (FRNT) in which a large number of serum samples can be handled at once with a small volume (15 L) of serum was useful for differential diagnosis between Japanese encephalitis and WN virus infections in infected chicks. Serological investigation was performed among wild birds in the Far Eastern region of Russia using the FRNT. Antibodies specific to WN virus were detected in 21 samples of resident and migratory birds out of 145 wild bird samples in the region.
  • 好井健太朗, 寸田祐嗣, 五十嵐学, 横澤香菜, 境瑞紀, 苅和宏明, HOLBROOK Michael 日本ウイルス学会学術集会プログラム・抄録集 61st 233 2013年10月29日 [査読無し][通常論文]
  • 好井健太朗, 寸田祐嗣, 五十嵐学, 横澤香菜, 境瑞紀, 苅和宏明, HOLBROOK Michael, 高島郁夫 日本獣医学会学術集会講演要旨集 156th 303 2013年08月30日 [査読無し][通常論文]
  • 【人と動物の共通感染症最前線10】 日本のげっ歯類におけるハンタウイルス感染の血清疫学調査とエゾヤチネズミが保有するHokkaidoウイルスの分離
    苅和 宏明, 尾崎 由佳, 真田 崇弘, 池中 良徳, 石塚 真由美, 坪田 敏男, 好井 健太朗, 吉松 組子, 有川 二郎, 高島 郁夫 獣医畜産新報 66 (4) 262 -264 2013年04月 [査読無し][通常論文]
     
    ハンタウイルスはげっ歯類媒介性の人獣共通感染症の病原体で、腎症候性出血熱(HFRS)またはハンタウイルス肺症候群(HPS)を引き起こす。近年の日本におけるげっ歯類のハンタウイルス感染状況を明らかにするために、1994年から2010年にかけて国内の様々な地域で捕獲された1658匹のげっ歯類の血清について、抗ハンタウイルス抗体の検出を行った。840例のRattus属げっ歯類(ドブネズミとクマネズミ)は全て抗体陰性であった。北海道以外の地域で捕獲された野生げっ歯類113例はいずれも抗体陰性であったのに対し、北海道で捕獲された705例の野性げっ歯類のうち、エゾヤチネズミの7.4%(26/352)とアカネズミの1.2%(2/168)が抗体を保有していた。(著者抄録)
  • 駒 貴明, 吉松 組子, 垂石 みどり, 宮下 大輔, 遠藤 理香, 清水 健太, 安田 俊平, 天田 貴子, 瀬戸 隆弘, 村田 亮, 吉田 喜香, 苅和 宏明, 高島 郁夫, 有川 二郎 北海道醫學雜誌 = Acta medica Hokkaidonensia 88 (1) 35 -35 2013年01月01日 [査読無し][通常論文]
  • 中尾桃子, 真田崇弘, 佐々木宣哉, サーサ ンゴンダ, 好井健太朗, 亀山武志, 高岡晃教, 苅和宏明 日本ウイルス学会学術集会プログラム・抄録集 60th 429 2012年10月31日 [査読無し][通常論文]
  • 中尾桃子, 真田崇弘, 佐々木宣哉, SAASA Ngonda, 好井健太朗, 亀山武志, 高岡晃教, 苅和宏明 日本獣医学会学術集会講演要旨集 154th 265 2012年08月31日 [査読無し][通常論文]
  • Hiroaki Kariwa, Haruka Yoshida, Cornelio Sanchez-Hernandez, Maria de Lourdes Romero-Almaraz, Jose Alberto Almazan-Catalan, Celso Ramos, Daisuke Miyashita, Takahiro Seto, Ayako Takano, Masashi Totani, Ryo Murata, Ngonda Saasa, Mariko Ishizuka, Takahiro Sanada, Kentaro Yoshii, Kumiko Yoshimatsu, Jiro Arikawa, Ikuo Takashima VIRUS RESEARCH 163 (2) 486 -494 2012年02月 [査読無し][通常論文]
     
    A variety of hantaviruses are harbored by rodents in North and South America, some of which can cause hantavirus pulmonary syndrome. To obtain greater evolutionary insight into hantaviruses in the Americas, a total of 211 rodents were captured in the Mexican states of Guerrero and Morelos in 2006. Anti-hantavirus antibodies were detected in 27 of 211 serum samples (12.8%) by ELISA. The distribution of seropositive rodents was: 17 Peromyscus beatae, 1 Megadontomys thomasi,1 Neotoma picta, 6 Reithrodontomys sumichrasti, and 2 Reithrodontomys megalotis. The hantavirus small (S), medium (M), and large (L) genome segments from P. beatae, R. sumichrasti, and R. megalotis were amplified and the sequences covering the open reading frames were determined. The hantaviruses from P. beatae, R. sumichrasti, and R. megalotis were provisionally designated Montano (MTN), Carrizal (CAR), and Huitzilac (HUI), respectively. The M segment amino acid identities among the Mexican hantaviruses were 80.8-93.0%. When these M segments were compared to those of known hantaviruses, MTN virus was most closely related to Limestone Canyon (LSC) virus (88.9% amino acid identity), while the CAR and HUI viruses were most closely related to El Moro Canyon (ELMC) virus (90-91% identity). Phylogenetic analysis revealed that the MTN, CAR, and HUI viruses occupy a monophyletic clade with the LSC, ELMC, and Rio Segundo viruses, which are harbored by Peromyscus boylii, R. megalotis, and Reithrodontomys mexicanus, respectively. The data obtained in this study provide important information for understanding the evolution of hantaviruses in the Americas. (C) 2011 Elsevier B.V. All rights reserved.
  • Takahiro Seto, Noriyo Nagata, Keisuke Yoshikawa, Osamu Ichii, Takahiro Sanada, Ngonda Saasa, Yuka Ozaki, Yasunori Kon, Kentaro Yoshii, Ikuo Takashima, Hiroaki Kariwa VIRUS RESEARCH 163 (1) 284 -290 2012年01月 [査読無し][通常論文]
     
    Hantaan virus (HTNV) is a causative agent of hemorrhagic fever with renal syndrome (HFRS). The pathogenesis of HFRS has not been fully elucidated, mainly due to the lack of a suitable animal model. In laboratory mice, HTNV causes encephalitis. However, that symptom is dissimilar to human hantavirus infections. We found that HTNV strain AA57 (isolated from Apodemus agrarius in Far East Russia) caused pulmonary disease in 2-week-old ICR mice. The clinical signs of the infected mice were piloerection, trembling, hunching, labored breathing, and body-weight loss. A large volume of pleural effusion was collected from thoracic cavities of the dead mice. Overall, 45% of the mice inoculated with 3000 focus forming units (FFU) of the virus began to show clinical symptoms at 8 days post-inoculation, and 25% of the inoculated mice died within 3 days of onset of the disease. The morbidity and mortality rates of the mice inoculated with 30-30,000 FFU of HTNV strain AA57 were roughly equivalent. The highest rates of virus positivity (11/12) and the highest titers of HTNV strain AA57 were detected in the lungs of the dead mice, while lower detection rates and viral titers were found in the heart, kidneys, spleen, and brain. Interstitial pneumonia, perivascular edema, hemorrhage, inflammatory infiltration and vascular failure were observed in the lungs of the sick mice. Hantaviral antigens were detected in the lung endothelial cells of the sick mice. The symptoms and pathology of this mouse model resemble those of hantavirus pulmonary syndrome (HPS) and, to a certain extent, those of HFRS. This is the first report that, in laboratory mice, the HFRS-related hantavirus causes a HPS-like disease and shares some symptom similarities with HFRS. (C) 2011 Elsevier B.V. All rights reserved.
  • Kentaro Yoshii, Manabu Igarashi, Osamu Ichii, Kana Yokozawa, Kimihito Ito, Hiroaki Kariwa, Ikuo Takashima JOURNAL OF GENERAL VIROLOGY 93 (1) 27 -38 2012年01月 [査読無し][通常論文]
     
    Flaviviruses are assembled to bud into the lumen of the endoplasmic reticulum (ER) and are secreted through the vesicle transport pathway, but the details of the molecular mechanism of virion assembly remain largely unknown. In this study, a highly conserved region in the prM protein was identified among flaviviruses. In the subviral particle (SP) system of tick-borne encephalitis virus (TBEV) and Japanese encephalitis virus, secretion of SPs was impaired by a mutation in the conserved region in the prM protein. Viral proteins were sparse in the Golgi complex and accumulated in the ER. Ultrastructural analysis revealed that long filamentous structures, rather than spherical SPs, were observed in the lumen of the ER as a result of the mutation. The production of infectious virions derived from infectious cDNA of TBEV was also reduced by mutations in the conserved region. Molecular modelling analysis suggested that the conserved region is important for the association of prM-envelope protein heterodimers in the formation of a spike of immature virion. These results are the first demonstration that the conserved region in the prM protein is a molecular determinant for the flavivirus assembly process.
  • Masashi Totani, Kentaro Yoshii, Hiroaki Kariwa, Ikuo Takashima AVIAN DISEASES 55 (4) 561 -568 2011年12月 [査読無し][通常論文]
     
    Birds are important for the transmission of West Nile virus (WNV) in nature, but the significance of the potential N-linked glycosylation at position 154 in the WNV envelope (E) protein with regard to viral replication in young chickens has not been assessed. In this study, the effect of glycosylation of the WNV E protein on viral pathogenicity in birds was investigated using young domestic chicks. A higher viral load was detected in the blood and the peripheral organs, particularly the hearts, of 2-day-old chicks inoculated with a glycosylated WNV variant compared to those inoculated with the nonglycosylated variant. There was no significant difference in the neutralizing antibody titers and cytokine expression profiles in chickens inoculated with the glycosylated and the nonglycosylated WNV variants. In contrast, no virus was detected in the blood and the tissues of 3-wk-old chicks, although the host immune response was induced to similar levels as in the 2-day-old chicks. These data indicate the utility of young domestic chicks as an animal model of WNV infection; they also indicate that glycosylation of the E protein of WNV enhances multiplication in the blood and peripheral organs, which is associated with the strong pathogenicity of WNV in birds.
  • Yuki Omori-Urabe, Kentaro Yoshii, Ayae Ikawa-Yoshida, Hiroaki Kariwa, Ikuo Takashima MICROBIOLOGY AND IMMUNOLOGY 55 (12) 893 -897 2011年12月 [査読無し][通常論文]
     
    Tick-borne encephalitis virus (TBEV) causes severe encephalitis in humans. It is endemic in one area of Japan; however no commercial vaccine is available in that country. In this Japan-based study, the efficacy of subviral particles (SPs) of TBEV administered by needle-free injector was evaluated as a vaccine candidate. Inoculation with SP-encoding DNA by needle-free injector induced neutralizing antibodies more efficiently than when administered by needle and syringe, and mice vaccinated with one dose by needle-free injector survived challenge with a lethal dose of TBEV. These results suggest that SP vaccines delivered by needle-free injector can protect against TBEV infection.
  • Ayako Takano, Kentaro Yoshii, Yuki Omori-Urabe, Kana Yokozawa, Hiroaki Kariwa, Ikuo Takashima ARCHIVES OF VIROLOGY 156 (11) 1931 -1941 2011年11月 [査読無し][通常論文]
     
    Tick-borne encephalitis virus (TBEV) causes severe encephalitis in humans. The Sofjin-HO strain is the prototype strain of the TBEV Far-Eastern subtype and is highly pathogenic in a mouse model. In this study, we constructed replicons and infectious cDNA clones of the Sofjin-HO strain. The replication of the replicon RNA was confirmed, and infectious viruses were recovered from the infectious cDNA clone. The recombinant viruses showed similar virulence characteristics to those of the parental virus. While characterizing the replicon and infectious cDNA, several amino acid differences derived from cell culture adaptations were analysed. The amino acids differences at E position 496 and NS4A position 58 were found to affect viral replication. The Gly- or Ala-to-Glu substitution at E position 122 was shown to increase neuroinvasiveness in mice. These replicons and infectious cDNA clones are useful in revealing the viral molecular determinants involved in the replication and pathogenicity of TBEV.
  • Takahiro Sanada, Hiroaki Kariwa, Noriyo Nagata, Yoichi Tanikawa, Takahiro Seto, Kumiko Yoshimatsu, Jiro Arikawa, Kentaro Yoshii, Ikuo Takashima VIRUS RESEARCH 160 (1-2) 108 -119 2011年09月 [査読無し][通常論文]
     
    The mechanism of hantavirus persistent infection in natural hosts is poorly understood due to a lack of laboratory animal models. Herein, we report that Syrian hamsters (Mesocricetus auratus) infected with Puumala virus (PUUV) at 4 weeks old show persistent infection without clinical symptoms for more than 2 months. IgG and IgM antibodies against the viral nucleocapsid protein and neutralizing antibody were first detectable at 14 days postinoculation (dpi) and maintained through 70 dpi. Viral RNA was first detected from 3 dpi in lungs and blood clots, and was detected in all tissues tested at 7 dpi. The viral RNA persisted for at least 70 days in the lungs, kidney, spleen, heart, and brain. The highest level of RNA copies was observed at 14 dpi in the lungs. Slight inflammatory reactions were observed in the lungs, adrenal glands, and brain. Immunohistochemical analysis revealed that PUUV antigen persisted until 56 dpi in the kidneys and adrenal glands. Infected hamsters showed no body weight loss or clinical signs. These results indicate that PUUV infection in hamsters is quite similar to the hantavirus infection of natural host rodents. (C) 2011 Elsevier B.V. All rights reserved.
  • 尾崎由佳, 萩谷友洋, 真田崇弘, 瀬戸隆弘, TAYLOR Kyle, 吉川佳佑, IVANOV Leonid, 好井健太朗, 坪田敏男, 池中良徳, 石塚真由美, 吉松組子, 有川二郎, 苅和宏明 日本獣医学会学術集会講演要旨集 152nd 256 2011年08月31日 [査読無し][通常論文]
  • 尾崎由佳, 萩谷友洋, 真田崇弘, 瀬戸隆弘, TAYLOR Kyle, 吉川佳佑, IVANOV Leonid I, 好井健太朗, 坪田敏男, 池中良徳, 石塚真由美, 吉松組子, 有川二郎, 苅和宏明 北海道獣医師会雑誌 55 (8) 415 2011年08月10日 [査読無し][通常論文]
  • Mayumi Obara, Takeo Yamauchi, Mamoru Watanabe, Sumiyo Hasegawa, Yasufumi Ueda, Kentaro Matsuno, Masae Iwai, Eiji Horimoto, Takeshi Kurata, Takenori Takizawa, Hiroaki Kariwa, Ikuo Takashima AMERICAN JOURNAL OF TROPICAL MEDICINE AND HYGIENE 84 (5) 695 -708 2011年05月 [査読無し][通常論文]
     
    To determine the mechanisms of maintenance and evolution of Japanese encephalitis virus (JEV) in a temperate zone, we attempted to isolate JEV from mosquitoes and pigs in Toyama Prefecture, Japan. A total of 87 JEVs were isolated from female Culex tritaeniorhynchus mosquitoes and pigs during 2005-2009. The prevalence of JEV in Toyama Prefecture was seasonally late in comparison with that of the virus during 1966-1972. Furthermore, JEVs were isolated after the peak in the number of female Cx. tritaeniorhynchus. Among JEV strains isolated in this study, two distinct groups were observed within genotype I of the phylogeny generated from nucleotide sequence information derived from the envelope and capsid/premembrane genes: strains belonging to the major type were isolated during 2005-2009, and strains from the minor type were isolated only in 2007. The major type has exhibited gradual change in its envelope and capsid/premembrane genes, and all isolates obtained in 2008 and 2009 had a novel deletion of seven nucleotides in the variable region of the 3'-untranslated region.
  • Takahiro Seto, Evgeniy A. Tkachenko, Vyacheslav G. Morozov, Yoichi Tanikawa, Sergey I. Kolominov, Sergey N. Belov, Ichiro Nakamura, Nobuo Hashimoto, Yasuhiro Kon, Alexander E. Balakiev, Tamara K. Dzagurnova, Olga A. Medvedkina, Mina Nakauchi, Mariko Ishizuka, Kentaro Yoshii, Kumiko Yoshimatsu, Leonid V. Ivanov, Jiro Arikawa, Ikuo Takashima, Hiroaki Kariwa JOURNAL OF VIROLOGICAL METHODS 173 (1) 17 -23 2011年04月 [査読無し][通常論文]
     
    Puumala virus (PUUV) and other Arvicolinae-borne hantaviruses are difficult to cultivate in cell culture. To isolate these hantaviruses efficiently, hantavirus nucleocapsid protein (NP)-positive but seronegative wild rodents were selected by NP-detection ELISA. Three of 68 Myodes glareolus captured in Samara, Russia, were NP-positive and seronegative. Syrian hamsters were inoculated with lung homogenates from NP-positive rodents for virus propagation. Virus isolation in vitro was carried out by inoculation of lung homogenates of NP-positive hamsters to Vero E6 cell monolayers. Two PUUV strains (Samara49/CG/2005 and Samara94/CG/2005) from M. glareolus were isolated in Vero E6 cells. Nucleotide and amino acid sequence identities of the S segment of these isolates to those of PUUV F-s808 from a fatal HFRS patient in Samara region were 96.7-99.3% and 99.3-100.0%, respectively. Morphologic features of Vero E6 cells infected with PUUV strain Samara49/CG/2005 were quite similar to those of Hantaan virus-infected cells. Isolation of Hokkaido virus from Myodes rufocanus captured in Hokkaido, Japan, was also performed. Hokkaido virus NP and RNA were recovered and maintained in hamsters. These results suggest that inoculation of Syrian hamsters with rodent samples is an efficient method for the isolation and maintenance of PUUV and other Arvicolinae-borne hantaviruses. (C) 2011 Elsevier B.V. All rights reserved.
  • 瀬戸隆弘, 吉川佳佑, 真田崇弘, SAASA Ngonda, 尾崎由佳, 市居修, 好井健太朗, 昆泰寛, 苅和宏明 日本獣医学会学術集会講演要旨集 151st 245 -245 2011年03月01日 [査読無し][通常論文]
  • Ryo Murata, Kazuaki Hashiguchi, Kentaro Yoshii, Hiroaki Kariwa, Kensuke Nakajima, Leonid I. Ivanov, Galina N. Leonova, Ikuo Takashima AMERICAN JOURNAL OF TROPICAL MEDICINE AND HYGIENE 84 (3) 461 -465 2011年03月 [査読無し][通常論文]
     
    West Nile (WN) virus has been spreading geographically to non-endemic areas in various parts of the world. However, little is known about the extent of WN virus infection in Russia. Japanese encephalitis (JE) virus, which is closely related to WN virus, is prevalent throughout East Asia. We evaluated the effectiveness of a focus reduction neutralization test in young chicks inoculated with JE and WN viruses, and conducted a survey of WN infection among wild birds in Far Eastern Russia. Following single virus infection, only neutralizing antibodies specific to the homologous virus were detected in chicks. The neutralization test was then applied to serum samples from 145 wild birds for WN and JE virus, Twenty-one samples were positive for neutralizing antibodies to WN. These results suggest that WN virus is prevalent among wild birds in the Far Eastern region of Russia.
  • Kentaro Yoshii, Keita Mottate, Yuki Omori-Urabe, Yumiko Chiba, Takahiro Seto, Takahiro Sanada, Junko Maeda, Mayumi Obara, Shuji Ando, Naoto Ito, Makoto Sugiyama, Hiroshi Sato, Hiroshi Fukushima, Hiroaki Kariwa, Ikuo Takashima JOURNAL OF VETERINARY MEDICAL SCIENCE 73 (3) 409 -412 2011年03月 [査読無し][通常論文]
     
    Tick-borne encephalitis virus (TBEV) is a zoonotic agent causing severe encephalitis in humans. Rodent species that are potential hosts for TBEV are widely distributed in various regions in Japan. In this study, we carried out large-scale epizootiological surveys in rodents from various areas of Japan. A total of 931 rodent and insectivore sera were collected from field surveys. Rodents seropositive for TBEV were found in Shimane Prefecture in Honshu and in several areas of Hokkaido Prefecture. These results emphasize the need for further epizootiological and epidemiological research of TBEV and preventive measures for emerging tick-borne encephalitis in Japan.
  • Ayae Ikawa-Yoshida, Kentaro Yoshii, Kazue Kuwahara, Mayumi Obara, Hiroaki Kariwa, Ikuo Takashima MICROBIOLOGY AND IMMUNOLOGY 55 (2) 100 -107 2011年02月 [査読無し][通常論文]
     
    Tick-borne encephalitis (TBE) virus causes severe encephalitis with serious sequelae in humans. An epizootiological survey of wild rodents is effective to detect TBE virus-endemic areas; however, limited serological diagnostic methods are available to detect anti-TBE virus antibodies in wild rodents. In this study, ELISAs for the detection of rodent antibodies against the TBE virus were developed using two recombinant proteins, domain III of the E protein (EdIII) and subviral particles (SPs), as the antigens. As compared with the neutralization test, the ELISA using EdIII had 77.1% sensitivity and 80.0% specificity, and the ELISA using SPs had 91.4% sensitivity and 100% specificity. Furthermore, when the ELISAs were applied to the epizootiological survey in the TBE virus-endemic area, both of the ELISAs was able to detect wild rodents with TBE virus-specific antibodies. This is the first study to show that ELISAs using recombinant antigens can be safe and useful in the detection of TBE virus-infected wild rodents in epizootiological research.
  • Kentaro Yoshii, Manabu Igarashi, Kimihito Ito, Hiroaki Kariwa, Michael R. Holbrook, Ikuo Takashima VIRUS RESEARCH 155 (1) 61 -68 2011年01月 [査読無し][通常論文]
     
    Omsk hemorrhagic fever virus (OHFV) is a member of the tick-borne encephalitis serocomplex of flaviviruses, and causes hemorrhagic disease in humans. In this study, an infectious cDNA of OHFV was constructed to investigate the molecular mechanisms involved in OHFV pathogenesis for the first time. Our cDNA clone was capable of producing infectious virus which is genetically identical to the parental Guriev strain, and the recombinant virus showed similar biological properties to the parental virus including growth kinetics and virulence characteristics. While characterizing the cDNAs, fortuitous mutations at NS2A position 46 and NS5 position 836 were found to affect viral production. By using a viral replicon expressing luciferase, it was shown that both of the mutations produced a defect in RNA replication and that the NS5 mutation induced a temperature-sensitive phenotype, indicating the importance of these residues in RNA replication. This infectious cDNA will be a useful tool to study the replication and pathogenesis of OHFV. (C) 2010 Elsevier B.V. All rights reserved.
  • Erdenesaikhan Tegshduuren, Kumiko Yoshimatsu, Midori Taruishi, Rika Endo, Kenta Shimizu, Takaaki Koma, Shumpei P. Yasuda, Hiroaki Kariwa, Jiro Arikawa, Chiaki Ishihara COMPARATIVE IMMUNOLOGY MICROBIOLOGY AND INFECTIOUS DISEASES 33 (6) E67 -E73 2010年12月 [査読無し][通常論文]
     
    Tula virus (TULV) and Puumala virus (PUUV) are hantaviruses carried by the bank vole (Myodes glareolus) and European common vole (Microtus arvalis), respectively. PUUV is a causative agent of hemorrhagic fever with renal syndrome (HFRS), while TULV is thought to be apathogenic to humans. The N-terminal regions of the N proteins from TULV and PUUV were expressed and applied as enzyme-linked immunosorbent assay (ELISA) antigens. Colonized Japanese grass voles (Microtus montebelli) and BALB/c mice were used for experimental inoculation of the vole-borne hantaviruses TULV and PUUV. Voles and mice showed significant antibody production toward both viruses, but these antisera showed little cross-reactivity between TULV and PUUV in the immunofluorescence antibody assay and ELISA. In contrast, sera from patients with HFRS caused by PUUV exhibited high cross-reactivity against the TULV antigen, and sera from a natural rodent reservoir showed moderate cross-reactivity against the heterologous antigen, indicating that the antigenic cross-reactivity between TULV and PUUV differs in sera from rodents and humans. (C) 2010 Elsevier Ltd. All rights reserved.
  • Hyun-Kyoung Lee, Byoung-Hee Lee, Seung-Hyeok Seok, Min-Won Baek, Hui-Young Lee, Dong-Jae Kim, Yi-Rang Na, Kyoung-Jin Noh, Sung-Hoon Park, Dutta Noton Kumar, Hiroaki Kariwa, Mina Nakauchi, Suk-Jin Heo, Jae-Hak Park JOURNAL OF VETERINARY SCIENCE 11 (2) 165 -167 2010年06月 [査読無し][通常論文]
     
    Severe acute respiratory syndrome (SARS) is a life-threatening disease for which accurate diagnosis is essential. Although many tools have been developed for the diagnosis of SARS, false-positive reactions in negative sera may occur because of cross-reactivity with other coronaviruses. We have raised polyclonal and monoclonal antibodies (Abs) using a recombinant form of the SARS virus nucleocapsid protein. Cross-reactivity of these anti-SARS Abs against human coronavirus (HCoV) 229E and HCoV OC43 were determined by Western blotting. The Abs produced reacted with recombinant SARS virus nucleocapsid protein, but not with HCoV 229E or HCoV OC43.
  • Ryo Murata, Yuki Eshita, Akihiko Maeda, Junko Maeda, Saki Akita, Tomohisa Tanaka, Kentaro Yoshii, Hiroaki Kariwa, Takashi Umemura, Ikuo Takashima AMERICAN JOURNAL OF TROPICAL MEDICINE AND HYGIENE 82 (4) 696 -704 2010年04月 [査読無し][通常論文]
     
    Many West Nile (WN) virus isolates associated with significant outbreaks possess a glycosylation site on the envelope (E) protein. E-protein glycosylated variants of New York (NY) strains of WN virus are more neuroinvasive in mice than the non-glycosylated variants. To determine how E protein glycosylation affects the interactions between WN virus and avian hosts, we inoculated young chicks with NY strains of WN virus containing either glycosylated or nonglycosylated variants of the E protein. The glycosylated variants were more virulent and had higher viremic levels than the non-glycosylated variants. The glycosylation status of the variant did not affect viral multiplication and dissemination in mosquitoes in vivo. Glycosylated variants showed more heat-stable propagation than non-glycosylated variants in mammalian (BHK) and avian (QT6) cells but not in mosquito (C6/36) cells. Thus, E-protein glycosylation may be a requirement for efficient transmission of WN virus from avian hosts to mosquito vectors.
  • Hiroaki Kariwa, Evgeniy A. Tkachenko, Vyacheslav G. Morozov, Takahiro Seto, Yoichi Tanikawa, Sergey I. Kolominov, Sergey N. Belov, Ichiro Nakamura, Nobuo Hashimoto, Alexander E. Balakiev, Tamara K. Dzagurnova, Nur Hardy bin Abu Daud, Daisuke Miyashita, Olga A. Medvedkina, Mina Nakauchi, Mariko Ishizuka, Kentaro Yoshii, Kumiko Yoshimatsu, Jiro Arikawa, Ikuo Takashima JOURNAL OF VETERINARY MEDICAL SCIENCE 71 (12) 1569 -1578 2009年12月 [査読無し][通常論文]
     
    European Russia is a highly endemic area of hemorrhagic fever with renal syndrome (HFRS), a rodent-borne zoonotic disease, caused by hantaviruses. In total, 145 small mammals of four species (Myodes glareolus, Apodemus flavicollis, A. agrarius, and A. uralensis) were trapped in the Samara region of European Russia in August 2005 and examined for the presence of hantavirus (HV). Anti-HV antibodies were found in six of 68 (8.8%) M. glareolus and in one of 19 (5.3%) A. flavicollis by indirect immunofluorescent antibody assay (IFA). The Puumala virus (PUUV), which is one of the hantavirus species, was detected in the lungs of seven M. glareolus by RT-PCR. The virus S-segment was extremely similar (96.2% to 99.3%) to the sequence found in a fatal case of HFRS in the Samara region. Phylogenetic analyses of S and M segments showed that the Samara PUUVs form a cluster within the Russian Volga lineage and apparently differ from other European PUUVs. Anti-PUUV antibodies were found in blood sera from seven HFRS patients and from one undiagnosed patient from the Samara region, using IFA and an enzyme-linked immunosorbent assay (ELISA). These data Suggest that the bank vole M. glareolus is a primary natural reservoir and vector for PUUV, which is the main causative agent of HFRS in humans in the Samara region.
  • Kentaro Yoshii, Ayae Ikawa, Yumiko Chiba, Yuki Omori, Junko Maeda, Ryo Murata, Hiroaki Kariwa, Ikuo Takashima JOURNAL OF VIROLOGICAL METHODS 161 (1) 173 -176 2009年10月 [査読無し][通常論文]
     
    Previously, a system for packaging tick-borne encephalitis virus (TBEV) subgenomic replicon RNAs into single-round infectious virus-like particles (VLPs) was developed. In the present study, VLPs were applied to measuring the levels of neutralizing antibodies against TBEV as an alternative to performing neutralization tests with live virus. As markers of VLP infection, the genes for GFP and luciferase were inserted into the TBEV replicon, which was then packaged into VLPs. The reporter genes were expressed in cells that were infected with the VLPs, and this infection was inhibited by neutralizing antibodies to TBEV. Serum samples from wild rodents were used to evaluate the neutralization test using VLPs. All the sera that were positive in the conventional neutralization test were also found to be positive in the neutralization test using VLPs, and there were highly significant correlations between the neutralization titres obtained using the native virus and those using VLPs. These results indicate that VLPs that express reporter genes represent a useful and safe alternative to conventional neutralization testing using live virus. (C) 2009 Elsevier B.V. All rights reserved.
  • Akihiko Maeda, Ryo Murata, Minoru Akiyama, Ikuo Takashima, Hiroaki Kariwa, Tomomasa Watanabe, Ichiro Kurane, Junko Maeda VIRUS RESEARCH 144 (1-2) 35 -43 2009年09月 [査読無し][通常論文]
     
    Viral reverse genetics, particularly infectious cloning, is a valuable tool with applications to many areas of viral research including the generation of vaccine candidates. However, this technology is sometimes insufficient for the construction cDNA clones as the genome sequences and/or encoding proteins of some viral agents may be toxic to the host cells used for cloning. To circumvent this problem, we developed a polymerase chain reaction (PCR)-based protocol for generating a complete West Nile virus (WNV) cDNA. The fragmented cDNAs were synthesized from WNV RNA by reverse transcription-PCR, and subsequently cloned into plasmids for use as templates for WNV cDNA synthesis. The fragmented cDNAs were amplified and assembled by PCR to generate a full-length WNV cDNA. Using this ONA as a template, WNV RNA was synthesized in vitro and transfected into mammalian cells. We also examined the generation of a mutant recombinant WNV containing a site-directed mutation within the viral genome sequence. Here, we discuss the possibility of developing a method for the generation of recombinant WNVs. (C) 2009 Elsevier B.V. All rights reserved.
  • 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 2009年08月 [査読無し][通常論文]
     
    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.
  • Mina Nakauchi, Hiroaki Kariwa, Yasuhiro Kon, Kentaro Yoshii, Akihiko Maeda, Ikuo Takashima MICROBIOLOGY AND IMMUNOLOGY 52 (12) 625 -630 2008年12月 [査読無し][通常論文]
     
    SARS-CoV has four major structural proteins: the N, S, M, and E proteins. To investigate the mechanism of SARS-CoV assembly, we cloned the genes encoding these four proteins into the eukaryotic expression vector pCAGGS and transfected them into 293T cells. When all four expression vectors were co-transfected VLP formed, as confirmed using electron microscopy. Using a rabbit polyclonal antibody specific to the N protein, N-protein-containing particles similar in size to the VLP were also observed by immunoelectron microscopy, indicating that the VLP contained the N protein. Co-immunoprecipitation analyses demonstrated an interaction between the N and M proteins, suggesting that N protein binds directly to M protein to be incorporated into VLP.
  • Nur Hardy Abu Daud, Hiroaki Kariwa, Evgeniy Tkachenko, Tamara Dzagurnova, Olga Medvedkina, Petr Tkachenko, Mariko Ishizuka, Takahiro Seto, Daisuke Miyashita, Takahiro Sanada, Mina Nakauchi, Kentaro Yoshii, Akihiko Maeda, Kumiko Yoshimatsu, Jiro Arikawa, Ikuo Takashima JAPANESE JOURNAL OF VETERINARY RESEARCH 56 (3) 151 -165 2008年11月 [査読無し][通常論文]
     
    Puumala virus (PUUV), a causative agent of hemorrhagic fever with renal syndrome (HFRS), is prevalent in Europe and European Russia. No vaccine has been developed for PUUV-associated HFRS, primarily because of the low viral yield in cultured cells. A PUUV strain known as DTK/Ufa-97 was isolated in Russia and adapted for growth in Vero E6 cells maintained in serum-free medium. The DTK/Ufa-97 strain produced a higher viral titer in serum-free medium, suggesting that it may prove useful in the development of an HFRS vaccine. When PUUV-infected Vero E6 cells were grown in serum-free medium, the DTK/Ufa-97 strain yielded more copies of intracellular viral RNA and a higher viral titer in the culture fluid than did the Sotkamo strain. Phylogenetic analysis revealed that PUUVs can be classified into multiple lineages according to geographical origin, and that the DTK/Ufa-97 strain is a member of the Bashkiria-Saratov lineage. The deduced amino acid sequences of the small, medium, and large segments of the DTK/Ufa-97 strain were 99.2% to 100%, 99.3% to 99.8%, and 99.8% identical, respectively, to those of the Bashkirian PUUV strains and 96.9%, 92.6%, and 97.4% identical, respectively, to those of the Sotkamo strain, indicating that the PUUVs are genetically diverse. However, DTK/Ufa-97 and other strains of PUUV exhibited similar patterns of binding to a panel of monoclonal antibodies against Hantaan virus. In addition, diluted antisera (i.e., ranging from 1:160 to 1:640) specific to three strains of PUUV neutralized both homologous and heterologous viruses. These results suggest that the DTK/Ufa-97 strain is capable of extensive growth and is antigenically similar to genetically distant strains of PUUV.
  • Hyun-Kyoung Lee, Byoung-Hee Lee, Noton Kumar Dutta, Seung-Hyeok Seok, Min-Won Baek, Hui-Young Lee, Dong-Jae Kim, Yi-Rang Na, Kyoung-Jin Noh, Sung-Hoon Park, Hiroaki Kariwa, Mina Nakauchi, Le Quynh Mai, Suk-Jin Heo, Jae-Hak Park JOURNAL OF MICROBIOLOGY AND BIOTECHNOLOGY 18 (10) 1717 -1721 2008年10月 [査読無し][通常論文]
     
    Severe acute respiratory syndrome (SARS) is a life-threatening emerging respiratory disease caused by the coronavirus, SARS-CoV The nucleocapsid (N) protein of SARS-CoV is highly antigenic and may be a suitable candidate for diagnostic applications. We constructed truncated recombinant N proteins (N1 |1-422 aa|, N2 |-109 aa|, and N3 |110-422 aa|) and determined their antigenicity by Western blotting using convalescent SARS serum. The recombinants containing N1 and N3 reacted with convalescent SARS serum in Western blotting. However, the recombinant with N2 did not. In ELISA using N1 or N3 as the antigens. positive results were observed in 10 of 10 (100%,) SARS-CoV-positive human sera. None of 50 healthy sera gave positive results in either assay. These data indicate that the ELISA using NI or N3 has high sensitivity and specificity. These results suggest that the middle or C-terminal region of the SARS N protein is important for eliciting antibodies against SARS-CoV during, the immune response, and ELISA reactions using N1 or N3 may be a valuable tool for SARS diagnosis.
  • Ichiro Nakamura, Kumiko Yoshimatsu, Byoung-Hee Lee, Megumi Okumura, Midori Taruishi, Koichi Araki, Hiroaki Kariwa, Ikuo Takashima, Jiro Arikawa ARCHIVES OF VIROLOGY 153 (8) 1537 -1542 2008年08月 [査読無し][通常論文]
     
    To distinguish Thailand virus infection from infections with other hantaviruses, we established an ELISA serotyping system using a truncated nucleocapsid protein of Thailand virus lacking 49 amino acids at the N-terminus. In evaluations using patient and rodent sera, Thailand virus infection was readily distinguished from Hantaan and Seoul virus infections. Therefore, this ELISA system is an effective alternative to neutralization tests.
  • Noton Kumar Dutta, Kaushiki Mazumdar, Byoung-Hee Lee, Min-Won Baek, Dong-Jae Kim, Yi-Rang Na, Sung-Hoon Park, Hyun-Kyoung Lee, Hiroaki Kariwa, Le Quynh Mai, Jae-Hak Park IMMUNOLOGY LETTERS 118 (1) 65 -71 2008年06月 [査読無し][通常論文]
     
    It is believed today that nucleocapsid protein (N) of severe acute respiratory syndrome (SARS)-CoV is one of the most promising antigen candidates for vaccine design. In this study, three fragments [NI (residues: 1-422); N2 (residues: 1-109); N3 (residues: 110-422)] of N protein of SARS-CoV were expressed in Escherichia coli and analyzed by pooled sera of convalescence phase of SARS patients. Three gene fragments [N1 (1-1269 nt), N2 (1-327 nt) and N3 (328-1269 nt)-expressing the same proteins of NI, N2 and N3, respectively] of SARS-N were cloned into pVAX-1 and used to immunize BALB/c mice by electroporation. Humoral (by enzyme-linked immunosorbent assay, ELISA) and cellular (by cell proliferation and CD4(+):CD8(+) assay) immunity was detected by using recombinant N1 and N3 specific antigen. Results showed that NI and N3 fragments of N protein expressed by E. coli were able to react with sera of SARS patients but N2 could not. Specific humoral and cellular immunity in mice could be induced significantly by inoculating SARS-CoV N1 and N3 DNA vaccine. In addition, the immune response levels in N3 were significantly higher for antibody responses (IgG and IgG1 but not IgG2a) and cell proliferation but not in CD4(+):CD8(+) assay compared to N1 vaccine. The identification of antigenic N protein fragments has implications to provide basic information for the design of DNA vaccine against SARS-CoV. The present results not only suggest that DNA immunization with pVax-N3 could be used as potential DNA vaccination approaches to induce antibody in BALB/c mice, but also illustrates that gene immunization with these SARS DNA vaccines can generate different immune responses. (c) 2008 Elsevier B.V. All rights reserved.
  • Hiroaki Kariwa, Hiroshi Noda, Mina Nakauchi, Mariko Ishizuka, Kazuaki Hashiguchi, Shingo Hashimoto, Kentaro Yoshii, Atsushi Asano, Takashi Agui, Hiroyuki Kogaki, Yoshihiro Kurano, Yoshiaki Uchida, Nobuyuki Fuji, Masahisa Okada, Ikuo Takashima JAPANESE JOURNAL OF VETERINARY RESEARCH 55 (4) 115 -127 2008年02月 [査読無し][通常論文]
     
    The sudden emergence of severe acute respiratory syndrome (SARS) at the end of 2002 resulted in 774 reported deaths from more than 8000 cases worldwide. As no effective vaccines or antiviral agents are available, the most effective measure to prevent the expansion of a SARS epidemic is the rapid diagnosis and isolation of SARS patients. To establish specific diagnostic methods, we generated nine clones of monoclonal antibodies to nucleocapsid protein (NP) of SARS-coronavirus (SARS-CoV). On immunofluorescent antibody assay and Western blotting analysis, none of the monoclonal antibodies showed cross-reactivity to authentic and recombinant NPs of human coronavirus (HCoV) 229E strain. To determine the region on the NP molecule where the monoclonal antibodies bind, we generated four truncated recombinant NPs and analyzed the reactivity between monoclonal antibodies and truncated NPs. Two monoclonal antibodies reacted with a truncated NP covering from amino acid residues 111 to 230, and seven reacted with another truncated NP covering from amino acid residues 221 to 340. Epitope mapping analysis indicated that monoclonal antibody SN5-25 recognized the amino acid sequence Q(245)TVTKW(250) on SARS-NP. Within the epitope, Q245, T246, V247, K249, and K250 appeared to form an essential motif for monoclonal antibody SN5-25 to bind. The information about binding sites and epitopes of monoclonal antibodies may be useful for the development of new diagnostic methods for SARS and for analyzing the function of N protein of SARS-CoV.
  • Kentaro Yoshii, Akiko Goto, Kazue Kawakami, Hiroaki Kariwa, Ikuo Takashima JOURNAL OF GENERAL VIROLOGY 89 (1) 200 -211 2008年01月 [査読無し][通常論文]
     
    We have previously reported a system for packaging tick-borne encephalitis (TBE) virus subgenomic replicon RNAs into single-round infectious virus-like particles (VLPs) by using in trans expression of viral C/prM/E structural proteins. In this study, the trans-packaging system was applied to the generation of chimeric VLPs with mosquito-borne Japanese encephalitis (JE) virus. Although trans-expression of TBE virus C and JE virus prM/E proteins resulted in the secretion of VLPs, the expression of JE virus C/prM/E proteins did not lead to the secretion of VLPs, suggesting that homologous interaction between C and non-structural proteins or the genomic RNA is important for efficient assembly of infectious particles. Neutralization testing showed that the antigenic characteristics of the VLPs; were similar to those of the native virus. Furthermore, the infectivities of the TBE virus- and JE virus-enveloped VLPs for the ISE6 tick cell line and C6/36 mosquito cell line were investigated. The VLPs were able to enter only those cells that were derived from the natural vectors for the respective viruses. TBE virus replicon RNA packaged in VLPs produced TBE virus non-structural proteins in tick cells, but could neither replicate nor produce viral proteins in mosquito cells. These findings indicate the importance of specific cellular factors for virus entry and replication during flavivirus infection of arthropods. These results demonstrate that chimeric VLPs are useful tools for the study of viral genome packaging and cellular factors involved in vector specificity, with the additional safety aspect that these chimeric VLPs can be used instead of full-length chimeric viruses.
  • Nur Hardy Abu Daud, Hiroaki Kariwa, Yoich Tanikawa, Ichiro Nakamura, Takahiro Seto, Daisuke Miyashita, Kentaro Yoshii, Mina Nakauchi, Kumiko Yoshimatsu, Jiro Arikawa, Ikuo Takashima MICROBIOLOGY AND IMMUNOLOGY 51 (11) 1081 -1090 2007年 [査読無し][通常論文]
     
    Hokkaido virus (HOKV) is a member of the genus Hantavirus, in the family Bunyaviridae. To investigate HOKV infection in the host Myodes rufocanus, the grey red-backed vole, 199 animals were captured at Tobetsu (October 2004 and July 2005) and Nakagawa (October 2004) in Hokkaido, Japan, for detection of antibody, antigen, and viral RNA. In the surveys in Tobetsu (2004) and Nakagawa (2004), seropositive animals were detected at a frequency of 6.0% (5/84) and 10.4% (5/48), respectively. No seropositive animals were detected in Tobetsu in 2005. Seroprevalence in males in Tobetsu and Nakagawa in 2004 was 25 % (1/4) and 45.5 % (5111), respectively, which was higher than in females, at 5.0 % (4/80) and 0% (0/37), respectively (P<0.01). These results suggest that male animals play an important role in the maintenance of HOKV in M. rufocanus. Two females were seronegative but viral RNA-positive, indicating that these animals had acute infections before antibody was produced. Another five infected animals in Nakagawa were all male and had high levels of antibodies and viral RNA, suggesting that they had persistent infections. Viral RNA copies in organs of infected animals in Nakagawa were quantified by real-time polymerase chain reaction. Two acutely infected animals had >= 10 times the number of RNA copies in their lungs compared to those of persistently infected animals. In most cases, lungs or spleen had the highest RNA copy number, regardless of infection status.
  • Sirima Pattamadilok, Byoung-Hee Lee, Sanit Kumperasart, Kumiko Yoshimatsu, Megumi Okumura, Ichiro Nakamura, Koichi Araki, Yuvaluk Khoprasert, Prayadh Dangsupa, Pornpitak Panlar, Burkhard Jandrig, Detlev H. Krueger, Boris Klempa, Thomas Jaekel, Jonas Schmidt, Rainer Ulrich, Hiroaki Kariwa, Jiro Arikawa AMERICAN JOURNAL OF TROPICAL MEDICINE AND HYGIENE 75 (5) 994 -1002 2006年11月 [査読無し][通常論文]
     
    Phylogenetic investigations, sequence comparisons, and antigenic cross-re activity studies confirmed the classification of Thailand virus (THAIV) as a distinct hantavirus species. The examination of sera from 402 rodents trapped in 19 provinces of Thailand revealed that five greater bandicoot rats (Bandicota indica) and one lesser bandicoot rat (B. savilei) from four provinces were focus reduction neutralization test (FRNT) antibody-positive for THAIV. One of 260 patients from Surin province in Thailand (initially suspected of having contracted leptospirosis, but found to be negative) showed symptoms compatible with hemorrhagic fever with renal syndrome (HFRS). The serum of this patient showed high titers of hantavirus-reactive IgM and IgG. FRNT investigations confirmed virus-neutralizing antibodies against THAIV. These observations suggest that THAIV or THAI-like viruses occur throughout Indochina and may represent an additional causative agent of HFRS.
  • Kazuya Shirato, Hirotsugu Miyoshi, Hiroaki Kariwa, Ikuo Takashima VIRUS RESEARCH 121 (1) 11 -16 2006年10月 [査読無し][通常論文]
     
    The envelope (E) protein glycosylation status of the New York strain of West Nile (WN) virus is an important determinant of virus neuroinvasiveness. To elucidate the determinant of the difference between E protein-glycosylated and non-glycosylated WN virus infections, the cytokine expression of murine peritoneal macrophages infected with each virus was examined. Tumor necrosis factor (TNF) alpha and interleukin (IL)-1 beta were up-regulated with replication of the E protein-glycosylated virus. Interferon (IFN) beta and IL-6 were up-regulated with the clearance of both viruses. These results suggest that TNF alpha and IL-1 beta expression are related to the virulence of E protein-glycosylated WN virus. (c) 2006 Elsevier B.V. All rights reserved.
  • M Obara, K Yoshii, T Kawata, D Hayasaka, A Goto, T Mizutani, H Kariwa, Takashima, I JOURNAL OF VIROLOGICAL METHODS 134 (1-2) 55 -60 2006年06月 [査読無し][通常論文]
     
    The similarity of symptoms produced by tick-borne encephalitis (TBE) and Japanese encephalitis (JE) and the high degree of cross-reactivity between TBE and JE viruses by serological tests make the development of a differential diagnostic test a priority. In this study, recombinant prM/E proteins of TBE virus strain Oshima 5-10 expressed in mammalian cells resulted in the release of subviral particles (SPs) into the culture medium. Using the SPs as antigens, enzyme-linked immunosorbent assay (ELISA) systems were developed to detect TBE virus-specific IgM and IgG antibodies, designated SP-IgG and SP-IgM ELISAs, respectively. Of 83 serum samples from encephalitis patients in Khabarovsk, Russia, which were positive with the neutralization test (NT), 82 were positive by the SP-IgG ELISA, for a sensitivity of 98.8%, which was higher than that of a commercial ELISA kit. All 12 NT-negative samples were also negative by the SP-IgG ELISA (specificity, 100%). Of 17 patient samples that were NT-positive, 16 (94.1%) were positive by the SP-IgM ELISA. Of 15 paired serum samples that yielded equivocal results by NT, 11 had positive results with the SP-IgM ELISA, indicating a diagnosis of TBE infection. The SP-IgG and SP-IgM ELISAs showed no cross-reactivity with antibodies to the JE virus. The results indicate that these ELISAs will be useful for the detection of TBE-specific antibodies. (c) 2005 Elsevier B.V. All rights reserved.
  • T Okabayashi, H Kariwa, S Yokota, S Iki, T Indoh, N Yokosawa, Takashima, I, H Tsutsumi, N Fujii JOURNAL OF MEDICAL VIROLOGY 78 (4) 417 -424 2006年04月 [査読無し][通常論文]
     
    The pathogenesis of severe acute respiratory syndrome (SARS) is poorly understood and cytokine dysregulation has been suggested as one relevant mechanism to be explored. We compared the cytokine profile in Caco2 cells after infection of SARS coronavirus (SARS-CoV) with other respiratory viruses including respiratory syncytial virus (RSV), influenza A virus (FluAV), and human parainfluenza virus type 2 (hPIV2). Interferon (IFN) system (production and response) was not suppressed by SARS-CoV infection. Therefore, SARS-CoV replication was suppressed by pretreatment with IFN. SARS-CoV and RSV induced high levels of IL-6 and RANTES compared with FluAV and hPIV2. Induction level of suppressor of cytokine signaling-3 (SOCS3) by SARS-CoV was significantly lower than that by RSV in spite of the significant production of IL-6. Toll-like receptors 4 and 9, which correlate with the induction of inflammatory response, were upregulated by SARS-CoV infection. Collectively, overinduction of inflammatory cytokine and dysregulation of cytokine signaling may contribute to the immunopathology associated with "severe" inflammation in SARS.
  • LJ Baek, H Kariwa, K Lokugamage, K Yoshimatsu, J Arikawa, Takashima, I, JI Kang, SS Moon, SY Chung, EJ Kim, HJ Kang, KJ Song, TA Klein, R Yanagihara, JW Song JOURNAL OF MEDICAL VIROLOGY 78 (2) 290 -297 2006年02月 [査読無し][通常論文]
     
    Hantaan (HTN) virus, the etiologic agent of clinically severe hemorrhagic fever with renal syndrome (HFRS), was first isolated in 1976 from lung tissue of a striped-field mouse (Apodemus agrarius) captured in Songnae-ri, Gyeonggi Province, Republic of Korea. Found primarily in mountainous areas, the Korean field mouse (A. peninsulae) is the second-most dominant field rodent species found throughout Korea. A new hantavirus, designated Soochong (SOO), was isolated in Vero E6 cells from four A. peninsulae captured in August 1997 at Mt. Gyebang in Hongcheon-gun, Mt. Gachil, Inje-gun, Gangwon Province, and in September 1998 at Mt. Deogyu, Muju-gun, Jeollabuk Province. The entire S, M, and L genomic segments of SOO virus, amplified by RT-PCR from lung tissues of seropositive A. peninsulae and from virus-infected Vero E6 cells, diverged from HTN virus (strain 76-118) by 15.6%, 22.8%, and 21.7% at the nucleotide level and 3.5%, 9.5%, and 4.6% at the amino acid level, respectively. Phylogenetic analyses of the nucleotide and deduced amino acid sequences, using the maximum parsimony and neighbor-joining methods, indicated that SOO virus was distinct from A. agrarius-borne HTN virus. SOO virus shared a common ancestry with Amur virus from Far East Russia, as well as with H5 and B78 hantaviruses, previously isolated from HFRS patients in China. Cross-focus-reduction neutralizating antibody tests showed that SOO virus which is the first hantavirus isolated in cell culture from A. peninsulae, could be classified as a new hantavirus serotype.
  • H Kariwa, N Fujii, K Takashima DERMATOLOGY 212 (Suppl 1) 119 -123 2006年 [査読無し][通常論文]
     
    The efficacy of several povidone-iodine (PVP-I) products, a number of other chemical agents and various physical conditions were evaluated for their ability to inactivate the severe acute respiratory syndrome coronavirus; (SARS-CoV). Treatment of SARS-CoV with PVP-I products for 2 min reduced the virus infectivity from 1.17 x 10(6) TCID50/ml to below the detectable level. The efficacy of 70% ethanol was equivalent to that of PVP-I products. Fixation of SARS-CoV-infected Vero E6 cells with a fixative including formalin, glutaraldehyde, methanol and acetone for 5 min or longer eliminated all infectivity. Heating the virus at 56 degrees C for 60 min or longer reduced the infectivity of the virus from 2.6 x 10(7) to undetectable levels. Irradiation with ultraviolet light at 134 mu W/cm(2) for 15 min reduced the infectivity from 3.8 x 10(7) to 180 TCID50/ml; however, prolonged irradiation (60 min) failed to eliminate the remaining virus, leaving 18.8 TCID50/ml. Copyright (c) 2006 S. Karger AG, Basel.
  • K Yoshii, D Hayasaka, A Goto, K Kawakami, H Kariwa, Takashima, I VACCINE 23 (30) 3946 -3956 2005年06月 [査読無し][通常論文]
     
    Yoshii, K., Hayasaka, D., Goto, A., Kawakami, K., Kariwa, H., and Takashima, I.:"Packaging the replicon RNA of the Far-Eastern subtype of tick-borne
    encephalitis virus into single-round infectious particles: development of a
    heterologous gene delivery system", Vaccine, 23:3946-3956.(2005)*
  • K Shirato, H Miyoshi, H Kariwa, Takashima, I JOURNAL OF VIROLOGICAL METHODS 126 (1-2) 119 -125 2005年06月 [査読無し][通常論文]
     
    A diagnostic method to distinguish between West Nile virus (WNV) and Japanese encephalitis virus (JEV) based on fluorogenic real-time polymerase chain reaction (TaqMan) assays was developed. To detect WNV and JEV with a single probe, a probe was designed to correspond to sequences in the core protein region that are shared by both viruses. The specificity of this assay depended on the primer sets used, which were specific to the target virus sequences: the primer set for WNV Could detect only WNV strains and the primer set for JEV could detect only JEV strains. The assays were tested by detection of viruses from experimentally infected animal tissues. The method described in this study will be useful for the simultaneous discrimination of WNV and JEV in areas where JEV is endemic, such as East Asia. © 2005 Elsevier B.V. All rights reserved.
  • Kogaki, H., Uchida, Y., Fujii, N., Kurano, Y., Miyake, K., Kido, Y., Kariwa, H., Takashima, I., Tamashiro, H., Ling, A.E., and Okada, M.:"Novel rapid immunochromatographic test based on an enzyme immunoassay for detecting nucleocapsid antigen in SARS-a・・・
    2005年 [査読無し][通常論文]
     
    Kogaki, H., Uchida, Y., Fujii, N., Kurano, Y., Miyake, K., Kido, Y., Kariwa, H., Takashima, I., Tamashiro, H., Ling, A.E., and Okada, M.:"Novel rapid immunochromatographic test based on an enzyme immunoassay for detecting nucleocapsid antigen in SARS-associated coronavirus", J Clin Lab Anal, 19:150-159. (2005)*
  • Goto, A., Yoshii, K., Obara, M., Ueki, T., Mizutani, T., Kariwa, H., and Takashima, I.:"Role of the N-linked glycans of the prM and E envelope proteins in tick-borne encephalitis virus particle secretion", Vaccine, 23:3043-3052(2005)*
    2005年 [査読無し][通常論文]
  • H Kariwa, N Fujii, Takashima, I JAPANESE JOURNAL OF VETERINARY RESEARCH 52 (3) 105 -112 2004年11月 [査読無し][通常論文]
     
    number of other chemical agents, and various physical conditions were evaluated for their ability to inactivate the severe acute respiratory syndrome coronavirus (SARS-CoV). Treatment of SARS-CoV with PVP-I products for 2 min reduced the virus infectivity from 1.17 x 10(6) TCID50/Ml to below the detectable level. The efficacy of 70% ethanol was equivalent to that of PVP-I products. Fixation of SARS-CoV-infected Vero E6 cells with a fixative including formalin, glutaraldehyde, methanol, and acetone for 5 min or longer eliminated all infectivity. Heating the virus at 56degreesC for 5 min dramatically reduced the infectivity of the virus from 2.6 x 10(7) to 40 TCID50/ml, whereas heating the virus for 60 min or longer eliminated all infectivity. Irradiation with ultraviolet light at 134mu W/cm(2) for 15 min reduced the infectivity from 3.8 x 10(7) to 180 TCID50/ml; however, prolonged irradiation (60 min) failed to eliminate the remaining virus, leaving 18.8 TCID50/ml. We believe that these findings will be useful for the implementation of infection control measures against SARS, and for the establishment of effective guidelines for the prevention of SARS outbreaks.
  • MA Iwasa, H Kariwa, BZ Cui, K Lokugamage, N Lokugamage, T Hagiya, T Mizutani, Takashima, I ARCHIVES OF VIROLOGY 149 (5) 929 -941 2004年05月 [査読無し][通常論文]
     
    Iwasa MA, Kariwa H, Cui BZ, Lokugamage K, Lokugamage N, Hagiya T, Mizutani T, Takashima I. Modes of hantavirus transmission in a population of Clethrionomys rufocanus bedfordiae inferred from mitochondrial and microsatellite DNA analyses. Arch Virol. 149(5): 929-941, 2004.*
  • Zamoto A, Tsuji M, Wei Q, Cho SH, Shin EH, Kim TS, Leonova GN, Hagiwara K, Asakawa M, Kariwa H, Takashima I, Ishihara C. Epizootiologic survey for Babesia microti among small wild mammals in northeastern Eurasia and a geographic diversity in the beta-t・・・
    2004年 [査読無し][通常論文]
     
    Zamoto A, Tsuji M, Wei Q, Cho SH, Shin EH, Kim TS, Leonova GN, Hagiwara K, Asakawa M, Kariwa H, Takashima I, Ishihara C. Epizootiologic survey for Babesia microti among small wild mammals in northeastern Eurasia and a geographic diversity in the beta-tubulin gene sequences. J Vet Med Sci. 66(7):785-792, 2004*
  • Shirato K, Miyoshi H, Goto A, Ako Y, Ueki T, Kariwa H, Takashima I. Viral envelope protein glycosylation is a molecular determinant of the neuroinvasiveness of the New York strain of West Nile virus. J Gen Virol. 85(Pt 12): 3637-3645, 2004.*
    2004年 [査読無し][通常論文]
  • Lokugamage K, Kariwa H, Lokugamage N, Iwasa M, Hagiya T, Araki K, Tachi A, Mizutani T, Yoshimatsu K, Arikawa J, Iwasaki T, Takashima I. Comparison of virulence of various hantaviruses related to hemorrhagic fever with renal syndrome in newborn mouse mo・・・
    2004年 [査読無し][通常論文]
     
    Lokugamage K, Kariwa H, Lokugamage N, Iwasa M, Hagiya T, Araki K, Tachi A, Mizutani T, Yoshimatsu K, Arikawa J, Iwasaki T, Takashima I. Comparison of virulence of various hantaviruses related to hemorrhagic fever with renal syndrome in newborn mouse model. Jpn J Vet Res. 51(3-4): 143-149, 2004.*
  • Araki K, Yoshimatsu K, Lee BH, Okumura M, Kariwa H, Takashima I, Arikawa J. Age-dependent hantavirus-specific CD8(+) T-cell responses in mice infected with Hantaan virus. Arch Virol. 149(7): 1373-1382, 2004.*
    2004年 [査読無し][通常論文]
  • Lokugamage N, Kariwa H, Lokugamage K, Iwasa MA, Hagiya T, Yoshii K, Tachi A, Ando S, Fukushima H, Tsuchiya K, Iwasaki T, Araki K, Yoshimatsu K, Arikawa J, Mizutani T, Osawa K, Sato H, Takashima I. Epizootiological and epidemiological study of hantavir・・・
    2004年 [査読無し][通常論文]
     
    Lokugamage N, Kariwa H, Lokugamage K, Iwasa MA, Hagiya T, Yoshii K, Tachi A, Ando S, Fukushima H, Tsuchiya K, Iwasaki T, Araki K, Yoshimatsu K, Arikawa J, Mizutani T, Osawa K, Sato H, Takashima I. Epizootiological and epidemiological study of hantavirus infection in Japan. Microbiol Immunol. 48(11): 843-851, 2004.*
  • Hayasaka D, Gritsun TS, Yoshii K, Ueki T, Goto A, Mizutani T, Kariwa H, Iwasaki T, Gould EA, Takashima I. Amino acid changes responsible for attenuation of virus neurovirulence in an infectious cDNA clone of the Oshima strain of tick-borne encephaliti・・・
    2004年 [査読無し][通常論文]
     
    Hayasaka D, Gritsun TS, Yoshii K, Ueki T, Goto A, Mizutani T, Kariwa H, Iwasaki T, Gould EA, Takashima I. Amino acid changes responsible for attenuation of virus neurovirulence in an infectious cDNA clone of the Oshima strain of tick-borne encephalitis virus. J Gen Virol. 85(Pt 4): 1007-1018, 2004.*
  • Lokugamage K, Kariwa H, Lokugamage N, Miyamoto H, Iwasa M, Hagiya T, Araki K, Tachi A, Mizutani T, Yoshimatsu K, Arikawa J, Takashima I. Genetic and antigenic characterization of the Amur virus associated with hemorrhagic fever with renal syndrome. Vir・・・
    2004年 [査読無し][通常論文]
     
    Lokugamage K, Kariwa H, Lokugamage N, Miyamoto H, Iwasa M, Hagiya T, Araki K, Tachi A, Mizutani T, Yoshimatsu K, Arikawa J, Takashima I. Genetic and antigenic characterization of the Amur virus associated with hemorrhagic fever with renal syndrome. Virus Res. 101(2): 127-134, 2004.*
  • Shirato K, Kimura T, Mizutani T, Kariwa H, Takashima I. Different chemokine expression in lethal and non-lethal murine West Nile virus infection. J Med Virol. 74(3): 507-513, 2004.*
    2004年 [査読無し][通常論文]
  • Yoshii K, Konno A, Goto A, Nio J, Obara M, Ueki T, Hayasaka D, Mizutani T, Kariwa H, Takashima I. Single point mutation in tick-borne encephalitis virus prM protein induces a reduction of virus particle secretion. J Gen Virol. 85(Pt 10):3049-3058, 20・・・
    2004年 [査読無し][通常論文]
     
    Yoshii K, Konno A, Goto A, Nio J, Obara M, Ueki T, Hayasaka D, Mizutani T, Kariwa H, Takashima I. Single point mutation in tick-borne encephalitis virus prM protein induces a reduction of virus particle secretion. J Gen Virol. 85(Pt 10):3049-3058, 2004.*
  • J Arikawa, K Yoshimatsu, H Wang, H Kariwa, K Lokugamage, N Lokugamage, Takashima, I JOURNAL OF CLINICAL VIROLOGY 28 S28 -S28 2003年12月 [査読無し][通常論文]
  • Mizutani, T., Kobayashi, M., Eshita, Y., Shirato, K., Kimura, T., Ako, Y., Miyoshi, H., Takasaki, T., Kurane, I., Kariwa, H., Umemura, T., and Takashima, I.:"Involvement of the JNK-like protein of the Aedes albopictus mosquito cell line, C6/36, in phag・・・
    2003年 [査読無し][通常論文]
     
    Mizutani, T., Kobayashi, M., Eshita, Y., Shirato, K., Kimura, T., Ako, Y., Miyoshi, H., Takasaki, T., Kurane, I., Kariwa, H., Umemura, T., and Takashima, I.:"Involvement of the JNK-like protein of the Aedes albopictus mosquito cell line, C6/36, in phagocytosis, endocytosis and infection of West Nile virus", Insect Molecular Biology, 12:491-499(2003)*
  • Goto, A., Hayasaka, D., Yoshii, K., Mizutani, T., Kariwa, H., and Takashima, I.:"A BHK-21 cell culture-adapted tick-borne encephalitis virus mutant is attenuated for neuroinvasiveness", Vaccine 21:4043-4051(2003)*
    2003年 [査読無し][通常論文]
  • Mizutani, T., Kobayashi, M., Eshita, Y., Inanami, O., Yamamori, T., Goto, A., Ako, Y., Miyoshi, H., Miyamoto, H., Kariwa, H., Kuwabara, M., and Takashima, I.:"Characterization of JNK-like protein derived from a mosquito cell line, C6/36", Insect Molec・・・
    2003年 [査読無し][通常論文]
     
    Mizutani, T., Kobayashi, M., Eshita, Y., Inanami, O., Yamamori, T., Goto, A., Ako, Y., Miyoshi, H., Miyamoto, H., Kariwa, H., Kuwabara, M., and Takashima, I.:"Characterization of JNK-like protein derived from a mosquito cell line, C6/36", Insect Molecular Biology, 12(1):61-66(2003)*
  • Kariwa, H., Tanabe, H., Mizutani, T., Kon, Y., Lokugamage, K., Lokugamage, N., Iwasa, M.A., Hagiya, T., Araki, K., Yoshimatsu, K., Arikawa, J., and Takashima, I.:"Synthesis of Seoul virus RNA and structural proteins in cultured cells", Archives of Viro・・・
    2003年 [査読無し][通常論文]
     
    Kariwa, H., Tanabe, H., Mizutani, T., Kon, Y., Lokugamage, K., Lokugamage, N., Iwasa, M.A., Hagiya, T., Araki, K., Yoshimatsu, K., Arikawa, J., and Takashima, I.:"Synthesis of Seoul virus RNA and structural proteins in cultured cells", Archives of Virology 148:1671-1685(2003)*
  • Lee, B.H., Yoshimatsu, K., Araki, K., Ogino, M., Okumura, M., Tsuchiya, K., Kariwa, H., and Arikawa, J.:"Detection of antibody for the serodiagnosis of hantavirus infection in different rodent species", Archives of Virology, 148:1885-1897(2003)*
    2003年 [査読無し][通常論文]
  • Shirato, K., Mizutani, T., Kariwa, H., and Takashima, I.:"Discrimination of West Nile virus and Japanese encephalitis virus strains using RT-PCR RFLP analysis", Microbiology and Immunology, 47:439-445(2003)*
    2003年 [査読無し][通常論文]
  • Miyamoto, H., Kariwa, H., Araki, K., Lokugamage, K., Hayasaka, D., Cui, B.Z., Lokugamage, N., Ivanov, L.I., Mizutani, T., Iwasa, M.A., Yoshimatsu, K., Arikawa, J., and Takashima, I.:" Serological analysis of hemorrhagic fever with renal syndrome (HFRS)・・・
    2003年 [査読無し][通常論文]
     
    Miyamoto, H., Kariwa, H., Araki, K., Lokugamage, K., Hayasaka, D., Cui, B.Z., Lokugamage, N., Ivanov, L.I., Mizutani, T., Iwasa, M.A., Yoshimatsu, K., Arikawa, J., and Takashima, I.:" Serological analysis of hemorrhagic fever with renal syndrome (HFRS) patients in Far Eastern Russia and identification of the causative hantavirus genotype", Archives of Virology, 148:1543-1556(2003)*
  • Araki, K., Yoshimatsu, K., Lee, B.H., Kariwa, H., Takashima, I., and Arikawa, J.:"Hantavirus-specific CD8(+)-T-cell responses in newborn mice persistently infected with Hantaan virus", Journal of Virology, 77:8408-8417(2003)*
    2003年 [査読無し][通常論文]
  • Lokugamage, N., Kariwa, H., Lokugamage, K., Hagiya, T., Miyamoto, H., Iwasa, M.A., Araki, K., Yoshimatsu, K., Arikawa, J., Mizutani, T., and Takashima, I.:"Development of an efficient method for recovery of Puumala and Puumala-related viruses by inocul・・・
    2003年 [査読無し][通常論文]
     
    Lokugamage, N., Kariwa, H., Lokugamage, K., Hagiya, T., Miyamoto, H., Iwasa, M.A., Araki, K., Yoshimatsu, K., Arikawa, J., Mizutani, T., and Takashima, I.:"Development of an efficient method for recovery of Puumala and Puumala-related viruses by inoculation of Mongolian gerbils", Journal of Veterinary Medical Science 65:1189-1194(2003)*
  • Yoshii, K., Hayasaka, D., Goto, A., Obara, M., Araki, K., Yoshimatsu, K., Arikawa, J., Ivanov, L., Mizutani, T., Kariwa, H., and Takashima, I.:"Enzyme-linked immunosorbent assay using recombinant antigens expressed in mammalian cells for serodiagnosis ・・・
    2003年 [査読無し][通常論文]
     
    Yoshii, K., Hayasaka, D., Goto, A., Obara, M., Araki, K., Yoshimatsu, K., Arikawa, J., Ivanov, L., Mizutani, T., Kariwa, H., and Takashima, I.:"Enzyme-linked immunosorbent assay using recombinant antigens expressed in mammalian cells for serodiagnosis of tick-borne encephalitis", Journal of Virological Methods, 108:171-179(2003)*
  • Y Yahara, Y Ohkubo, H Kariwa, Takashima, I JOURNAL OF VETERINARY MEDICAL SCIENCE 64 (7) 583 -588 2002年07月 [査読無し][通常論文]
     
    Yahara, Y., Ohkubo, Y., Kariwa, H., Takashima, I.:"Evaluation of enzyme-linked immunosorbent assay (ELISA) and immunofluorescent antibody (IFA) test for the detection of porcine reproductive and respiratory syndrome virus (PRRSV) antibody in pigs from conventional farms"
    J Vet Med Sci, 64:583-588(2002)*
  • Goto, A., Hayasaka, D., Yoshii, K., Mizutani, T., Kariwa, H., Takashima, I. :"Genetic and biological comparison of tick-borne encephalitis viruses from Hokkaido and far-eastern Russia": Jpn J Vet Res, 49:297-307(2002)*
    2002年 [査読無し][通常論文]
  • Lokugamage, K., Kariwa, H., Hayasaka, D., Cui, B.Z., Iwasaki, T., Lokugamage, N., Ivanov, L.I., Volkov, V.I., Demenev, V.A., Slonova, R., Kompanets, G., Kushnaryova, T., Kurata, T., Maeda, K., Araki, K., Mizutani, T., Yoshimatsu, K., Arikawa, J., Takas・・・
    2002年 [査読無し][通常論文]
     
    Lokugamage, K., Kariwa, H., Hayasaka, D., Cui, B.Z., Iwasaki, T., Lokugamage, N., Ivanov, L.I., Volkov, V.I., Demenev, V.A., Slonova, R., Kompanets, G., Kushnaryova, T., Kurata, T., Maeda, K., Araki, K., Mizutani, T., Yoshimatsu, K., Arikawa, J., Takashima, I.:"Genetic characterization of hantaviruses transmitted by the Korean field mouse (Apodemus peninsulae), Far East Russia"
    Emerg Infect Dis, 8:768-776(2002)*
  • H Wang, K Yoshimatsu, H Ebihara, M Ogino, K Araki, H Kariwa, ZX Wang, ZZ Luo, DX Li, CS Hang, J Arikawa VIROLOGY 278 (2) 332 -345 2000年12月 [査読無し][通常論文]
     
    The antigenic and genetic properties of 46 hantaviruses from China, 13 from patients, 23 from rodents, and 10 from unknown hosts, were compared with those of other hantaviruses. The viruses were classified as either Hantaan (HTN) or Seoul (SEO) viruses. A phylogenetic analysis of the partial M (300 bp) and S (around 485 bp) genomes of HTN viruses identified nine distinct genetic subtypes, one consisting of isolates from Korea. The SEC viruses were divided into five genetic subtypes, although they had less variability than the HTN subtypes. There was a correlation between the subtype and province of origin for four subtypes of HTN viruses, confirming geographical clustering. Hantaan virus NC167 isolated from Niviventer confucianus and SEO virus Gou3 isolated from Rattus rattus were the basal clades in each virus. The phylogenetic trees constructed from the entire S and M segments suggested that NC167 was introduced to N. confucianus in a host-switching event. The reactivity of a panel of 35 monoclonal antibodies was almost exactly the same in NC167 and a representative HTN virus and in Gou3 and a representative SEC virus. However. there was a one-way cross-neutralization between them. These results confirm the varied nature of Murinae-associated hantaviruses in China. (C) 2000 Academic Press.
  • H Ebihara, K Yoshimatsu, M Ogino, K Araki, Y Ami, H Kariwa, Takashima, I, DX Li, J Arikawa JOURNAL OF VIROLOGY 74 (19) 9245 -9255 2000年10月 [査読無し][通常論文]
     
    Ebihara, H., Yoshimatsu, K., Ogino, M., Araki, K., Ami, Y., Kariwa, H., Takashima I, Li, D. and Arikawa J.: "Pathogenicity of Hantaan virus in newborn mice: genetic reassortant study demonstrating that a single amino acid change in glycoprotein G1
    is related to virulence", Journal of Virology, 74:9245-9255(2000)*
  • Wang, H., Yoshimatsu, K., Ebihara, H., Ogino, M., Araki, K., Kariwa, H., Wang, Z., Luo, Z., Li, D., Hang, C. and Arikawa, J.: "Genetic diversity of hantaviruses isolated in china and characterization of novel hantaviruses isolated from Niviventer confu・・・
    2000年 [査読無し][通常論文]
     
    Wang, H., Yoshimatsu, K., Ebihara, H., Ogino, M., Araki, K., Kariwa, H., Wang, Z., Luo, Z., Li, D., Hang, C. and Arikawa, J.: "Genetic diversity of hantaviruses isolated in china and characterization of novel hantaviruses isolated from Niviventer confucianus and Rattus rattus", Virology, 278:332-345(2000)*
  • Kariwa, H., Yoshimatsu, K., Araki, K., Chayama, K., Kumada, H., Ogino, M., Ebihara, H., Murphy, M.E., Mizutani, T., Takashima, I. and Arikawa, J.: "Detection of hantaviral antibodies among patients with hepatitis of unknown etiology in Japan", Microbio・・・
    2000年 [査読無し][通常論文]
     
    Kariwa, H., Yoshimatsu, K., Araki, K., Chayama, K., Kumada, H., Ogino, M., Ebihara, H., Murphy, M.E., Mizutani, T., Takashima, I. and Arikawa, J.:
    "Detection of hantaviral antibodies among patients with hepatitis of unknown etiology in Japan", Microbiology and Immunology, 44:357-362(2000)*
  • Murphy, M.E., Kariwa, H., Mizutani, T., Yoshimatsu, K., Arikawa, J. and Takashima I.: "In vitro antiviral activity of lactoferrin and ribavirin upon hantavirus", Archives of Virology, 145:1571-1582(2000)*
    2000年 [査読無し][通常論文]
  • Tetsuya Mizutani, Hisae Inagaki, Mitsuhiro Tada, Daisuke Hayasaka, Michael Murphy, Toshiyoshi Fujiwara, Jun-Ichi Hamada, Hiroaki Kariwa, Ikuo Takashima Microbiology and Immunology 44 (7) 597 -603 2000年 [査読無し][通常論文]
     
    Mizutani, T., Inagaki, H., Tada, M., Hayasaka, D., Murphy, M., Fujiwara, T., Hamada, J., Kariwa, H. and Takashima, I.: "The mechanism of actinomycin D-mediated increase of Borna disease virus (BDV) RNA in cells persistently infected by BDV", Microbiology and Immunology, 44:597-603(2000)*
  • Komoro, K., Hayasaka, D., Mizutani, T., Kariwa, H. and Takashima, I.: "Characterization of monoclonal antibodies against Hokkaido strain tick-borne encephalitis virus", Microbiology and Immunology,44:533-536(2000)*
    2000年 [査読無し][通常論文]
  • H Kariwa, K Yoshimatsu, J Sawabe, E Yokota, J Arikawa, Takashima, I, H Fukushima, A Lundkvist, FN Shubin, LM Isachkova, RA Slonova, GN Leonova, N Hashimoto VIRUS RESEARCH 59 (2) 219 -228 1999年02月 [査読無し][通常論文]
     
    Seroepizootiologic surveys among wild rodents were carried out in Japan and Far East Russia in 1995 and 1996. Seropositive animals were only identified in Clethrionomys rufocanus (23/134) in Hokkaido, Japan. On the other hand, seropositives were identified in C. rufocanus (1/8), Apodemus argrarius (2/66), Apodemus spp. (2/26) and Microtus fortis (3/22) in Vladivostok, Far East Russia. Total RNA was isolated from lungs of seropositive animals and the S genome segments were amplified by PCR, cloned and sequenced. The S and M genomes of hantavirus, derived from Japanese C. rufocanus (Tobetsu genotype), were most closely related with Puumala viruses (76-79% nucleotide and 95% amino acid identities for S genome, 70-78% nucleotide and 87-92% amino acid identities for M genome). The recombinant nucleocapsid protein of Tobetsu genotype was antigenically quite similar with that of Sotkamo. These suggest that the virus endemic in Japanese C. rufocanus belongs to Puumala virus. Phylogenetic analysis indicates that the genotype forms a distinct lineage within Puumala viruses. Partial S segment (1-1251 nt), derived from seropositive M. fortis in Vladivostok, was sequenced and analyzed. The S genome segment, which was designated Vladivostok genotype, was most closely related with Khabarovsk virus (79% nucleotide and 90% amino acid identities) which was isolated from M. fortis. (C) 1999 Elsevier Science B.V. All rights reserved.
  • Kariwa H, Yoshimatsu K, Sawabe J, Yokota E, Arikawa J, Takashima I, Fukushima H, Lundkvist A, Shubin FN, Isachkova LM, Slonova RA, Leonova GN, Hashimoto N.:"Genetic diversities of hantaviruses among rodents in Hokkaido, Japan and Far East Russia", Viru・・・
    1999年 [査読無し][通常論文]
     
    Kariwa H, Yoshimatsu K, Sawabe J, Yokota E, Arikawa J, Takashima I, Fukushima H, Lundkvist A, Shubin FN, Isachkova LM, Slonova RA, Leonova GN, Hashimoto N.:"Genetic diversities of hantaviruses among rodents in Hokkaido, Japan and Far East Russia", Virus Res., 59(2):219-28 (1999)*
  • Hayasaka D, Suzuki Y, Kariwa H, Ivanov L, Volkov V, Demenev V, Mizutani T, Gojobori T, Takashima I.:"Phylogenetic and virulence analysis of tick-borne encephalitis viruses from Japan and far-Eastern Russia", J Gen Virol., 80 (12):3127-3135 (1999)*
    1999年 [査読無し][通常論文]
  • Mizutani T, Inagaki H, Hayasaka D, Kariwa H, Takashima I.:"Enhancement of Borna disease virus transcription in persistently infected cells by serum starvation", J Vet Med Sci., 61(7):831-834 (1999)*
    1999年 [査読無し][通常論文]
  • Chiba N, Osada M, Komoro K, Mizutani T, Kariwa H, Takashima I.:"Protection against tick-borne encephalitis virus isolated in Japan by active and passive immunization", Vaccine., 17(11-12):1532-1539 (1999)*
    1999年 [査読無し][通常論文]
  • T. Mizutani, H. Inagaki, D. Hayasaka, S. Shuto, N. Minakawa, A. Matsuda, H. Kariwa, I. Takashima Archives of Virology 144 (10) 1937 -1946 1999年 [査読無し][通常論文]
     
    Regulation of viral RNA levels in infected cells is considered important in the investigation of viral transcription and replication. Amounts of Borna disease virus (BDV) RNAs were increased in confluent persistently BDV-infected MDCK cells (MDCK/BDV) cells, while maintained at low levels in growing cells. The amount of 1.9-kb RNA without cap formation and polyadenylation at the 5' and 3' ends respectively were remarkably increased (200% per day) in confluent MDCK/BDV cells. Both the full-length genomic and anti-genomic RNAs were increased accompanied by 1.9-kb RNA, suggesting the transcription of the 1.9-kb RNA was important for replication of BDV. Ribavirin has an inhibitory effect on replication and transcription of BDV at concentrations from 1 to 10 μg/ml [Mizutani T et al., Arch Virol (1998)143:2039-2044]. BDV transcripts were decreased with ribavirin treatment and increased after its removal which indicated that ribavirin has a reversible inhibitory effect on BDV transcription. Furthermore, BDV transcription was also decreased by two agents, RMNPA and EICAR, which selectively inhibit enzyme activity related to cap formation at the 5' end of mRNA. On the contrary, when the growing MDCK/BDV cells were treated with actinomycin D, transcripts of BDV RNA were increased for 24 h. These agents and culture conditions in this study were found to be useful tools for up- and down-regulation of BDV transcription in persistently BDV-infected cells.
  • Mizutani T, Nishino Y, Kariwa H, Takashima I.:"Reverse transcription-nested polymerase chain reaction for detecting p40 RNA of Borna disease virus, without risk of plasmid contamination", J Vet Med Sci., 61(1):77-80 (1999)*
    1999年 [査読無し][通常論文]
  • Mizutani T, Ogino M, Nishino Y, Kimura T, Inagaki H, Hayasaka D, Kariwa H, Takashima I.:"Single-step reverse transcriptase-polymerase chain reaction for detection of Borna disease virus RNA in vitro and in vivo", Jpn J Vet Res., 46(4):165-169 (1999)*
    1999年 [査読無し][通常論文]
  • Tsujimura K, Mizutani T, Kariwa H, Yoshimatsu K, Ogino M, Morii Y, Inagaki H, Arikawa J, Takashima I.:"A serosurvey of Borna disease virus infection in wild rats by a capture ELISA", J Vet Med Sci., 61(2):113-7 (1999)*
    1999年 [査読無し][通常論文]
  • Chiba N, Iwasaki T, Mizutani T, Kariwa H, Kurata T, Takashima I.:"Pathogenicity of tick-borne encephalitis virus isolated in Hokkaido, Japan in mouse model", Vaccine., 17(7-8):779-787 (1999)*
    1999年 [査読無し][通常論文]
  • H Kariwa, M Fujiki, K Yoshimatsu, J Arikawa, Takashima, I, N Hashimoto ARCHIVES OF VIROLOGY 143 (1) 15 -24 1998年 [査読無し][通常論文]
     
    To understand the mode of transmission of Seoul type hantavirus in Wistar rats, we examined the shedding of the virus and antibody production in infected rats. When 1-day-old rats were inoculated with the KI-83-262 strain of Seoul virus, S segment of the viral genome was detected in lungs, clots, urine, saliva, submaxillary glands, rectums, and kidneys by nested reverse transcriptase PCR. On the other hand, when 8-week-old rats were infected with the virus, viral genome was detected only in the lungs and rectum. In newborn rats intranasally administered urine from infected newborn rats, four of six rats shed the virus into their urine. In addition, three of eight rats kept in the same cage with infected animals also shed the virus into urine. Moreover, the virus genome was detected in the urine of urban rats (Rattus norvegicus) in an enzootic focus. These findings suggest that the urine containing virus from infected rats is an actual source of the Seoul virus infection.
  • *Kariwa, H., Fujiki, M., Yoshimatsu, K., Arikawa, J., Takashima, I., Hashimoto, N. "Urine-associated horizontal transmission of Seoul virus among rats", Archives of Virology, 143: 15-24 (1998)*
    1998年 [査読無し][通常論文]
  • Morii, M., Yoshimatsu, K., Arikawa, J., Zhou, G, Kariwa, H., Takashima, I. "Antigenic characterization of Hantaan and Seoul virus nucleocapsid proteins expressed by recombinant baculovirus: application of a truncated protein, lacking an antigenic regio・・・
    1998年 [査読無し][通常論文]
     
    Morii, M., Yoshimatsu, K., Arikawa, J., Zhou, G, Kariwa, H., Takashima, I. "Antigenic characterization of Hantaan and Seoul virus nucleocapsid proteins expressed by recombinant baculovirus: application of a truncated protein, lacking an antigenic region common to the two viruses, as a serotyping antigen", Journal of Clinical Microbiology, 36: 2514-2521 (1998) *
  • Takashima, I, K Morita, M Chiba, D Hayasaka, T Sato, C Takezawa, A Igarashi, H Kariwa, K Yoshimatsu, J Arikawa, N Hashimoto JOURNAL OF CLINICAL MICROBIOLOGY 35 (8) 1943 -1947 1997年08月 [査読無し][通常論文]
     
    A case of tick-borne encephalitis (TEE) has not been reported for many years in Japan, although a serological survey of sera from domestic animals suggested the presence of TEE foci in Hokkaido, the northern island of Japan. Studies were conducted to prove the presence of an endemic focus of TEE virus in Japan by means of serology and virus isolation. In October 1993 in Hokkaido, a severe case of encephalitis in a dairy farmer's nife was diagnosed as TEE. Serological examination of paired serum specimens showed a rise in the neutralization antibody titer to Russian spring summer encephalitis virus. A seroepizootiological survey of dogs showed that the TEE-related virus was prevalent in the area. Three virus isolates were obtained from the blood of sentinel dogs, and antigenic analysis grouped the isolates into TEE-related viruses. Sequence analysis of the envelope protein gene identified one of the isolates as being of the same subtype as the Russian spring summer encephalitis (Far Eastern TEE) virus. The results provide evidence that TEE is endemic in a certain area of Japan.
  • Takashima, I., Morita, K., Chiba, M., Hayasaka, D., Sato, T., Takezawa, C., Igarashi, A., Kariwa, H., Yoshimatsu, K., Arikawa, J. and Hashimoto, N. : "A case of tick-borne encephalitis in Japan and isolation of the the virus", J. Clin. Microbiol., 35 :・・・
    1997年 [査読無し][通常論文]
     
    Takashima, I., Morita, K., Chiba, M., Hayasaka, D., Sato, T., Takezawa, C., Igarashi, A., Kariwa, H., Yoshimatsu, K., Arikawa, J. and Hashimoto, N. : "A case of tick-borne encephalitis in Japan and isolation of the the virus", J. Clin. Microbiol., 35 : 1943-1947 (1997)*
  • Ennis, F. A., Cruz, J., Spiropoulou, C. F., Waite, D., Peters, C. J., Nichol, S. T., Kariwa, H. and Koster, F. T. : "Hantavirus pulmonary syndrome : CD8+ and CD4+ cytotoxic T lymphocytes to epitopes on Sin Nombre virus nucleocapsid protein isolated dur・・・
    1997年 [査読無し][通常論文]
     
    Ennis, F. A., Cruz, J., Spiropoulou, C. F., Waite, D., Peters, C. J., Nichol, S. T., Kariwa, H. and Koster, F. T. : "Hantavirus pulmonary syndrome : CD8+ and CD4+ cytotoxic T lymphocytes to epitopes on Sin Nombre virus nucleocapsid protein isolated during acute illness", Virology, 238 : 380-390 (1997)*
  • A Ito, M Okamoto, H Kariwa, T Ishiguro, A Hashimoto, M Nakao JOURNAL OF HELMINTHOLOGY 70 (4) 355 -357 1996年12月 [査読無し][通常論文]
     
    Two Norway rats, Rattus norvegicus, were found to be naturally infected with Echinococcus multilocularis in Japan. One of them was simultaneously infected with at least three different sized metacestodes of Taenia taeniaeformis. These two R. norvegicus rats and another R. norvegicus naturally infected with T. taeniaeformis and Capillaria hepatica were examined to see if they showed any antibody responses against these two cestode parasites with the view to obtaining more information on the importance of rats as the intermediate host for E. multilocularis. These R. norvegicus showed very poor antibody responses against the two cestode species, although the Wistar rats, R. rattus, experimentally infected with a single smaller sized metacestode of T. taeniaeformis showed stronger responses not only against T. taeniaeformis but also against E. multilocularis. Therefore the three R. norvegicus naturally infected with E. multilocularis and/or T. taeniaeformis demonstrated virtually no immune response, at least against these cestodes.
  • K Yoshimatsu, J Arikawa, M Tamura, R Yoshida, A Lundkvist, B Niklasson, H Kariwa, Azuma, I JOURNAL OF GENERAL VIROLOGY 77 695 -704 1996年04月 [査読無し][通常論文]
     
    We characterized the antigenic sites on the nucleocapsid protein (NP) of Hantaan virus (HTN) using 10 monoclonal antibodies (MAbs). At least seven antigenic sites were revealed by a competitive binding assay and divided into three partially overlapping antigenic regions (I, II and III). Regions I [amino acids (aa) 1-103], II (aa 104-204) and III (aa 205-402) were mapped on NP by examinincr the reactivity of truncated gene products. Those that corresponded to region I reacted with immune mouse serum, indicating that the region contained major linear epitopes as reported with Four corners virus (FCV) and Puumala virus (PUU) NP. At least one MAb to each region inhibited viral growth when they were introduced into cells by scrape-loading. In addition, they conferred protection from a lethal HTN challenge to newborn mice. A PEPSCAN assay localized the epitope of MAb E5/G6 between aa 166-175. Since E5/G6, which had the highest inhibitory effect both in cells and in mice, showed no virus neutralization activity by ordinary neutralization test, this region is suggested to be important for the virus growth after entry into the cells.
  • K Yoshimatsu, J Arikawa, H Li, H Kariwa, N Hashimoto JOURNAL OF VETERINARY MEDICAL SCIENCE 58 (1) 71 -74 1996年01月 [査読無し][通常論文]
     
    K. Yoshimatsu, J. Arikawa, H. Li, H. Kariwa and N. Hashimoto : "Western blotting using recombinant Hantaan virus nucleocapsid protein expressed in silkworm as a serological confirmation of hantavirus infection in human sera", J. Vet. Med. Sci., 58 : 71-74 (1996)*
  • I. Takashima, M. Hiyoshi, H. Kariwa, R. Mukaiya and N. Hashimoto : "Experimental Chlamydia psittaci infection of Japanese quail", Microbiol. Immunol., 40 : 265-701 (1996)*
    1996年 [査読無し][通常論文]
  • I. Takashima, Y. Imai, N. Itoh, H. Kariwa and N. Hashimoto : "Polymerase chain reaction for the detection of Chlamydia psittaci in the feces of budgerigars", Microb. Immunol., 40 : 21-26 (1996)*
    1996年 [査読無し][通常論文]
  • H. Kariwa, M. Kimura, S. Yoshizumi, J. Arikawa, K. Yoshimatsu, I. Takashima and N. Hashimoto : "Different modes of Seoul virus infection : persistency in newborn rats and transiency in adult rats", Arch. Virol., 141 : 2327-2338 (1996)*
    1996年 [査読無し][通常論文]

書籍等出版物

  • Animal viruses
    Transworld Research Network 2010年
  • SARS
    Transworld Research Network 2006年
  • Hantaviruses and Hantavirus infections
    Times New Roman 2003年

担当経験のある科目(授業)

  • 畜産食品衛生学北海道大学
  • 人獣共通感染症学「北海道大学」「帯広畜産大学」
  • 獣医公衆衛生学北海道大学

所属学協会

  • 人と動物の共通感染症研究会   獣医疫学会   日本ウイルス学会   日本獣医学会   日本獣医公衆衛生学会   The Japan Society of Veterinary Epidemiology   The Japanese Society for Virology   The Japanese Society of Veterinary Science   The Japanese Society of Veterinary Public Health   

共同研究・競争的資金等の研究課題

  • 日本学術振興会:科学研究費助成事業 基盤研究(B)
    研究期間 : 2013年04月 -2017年03月 
    代表者 : 苅和 宏明, 有川 二郎, 好井 健太朗, 森松 組子
     
    本研究では、近年国内外で問題となっているウイルス性人獣共通感染症のうち、重篤化しやすく致死率も高いハンタウイルス感染症、ダニ媒介性脳炎、およびウエストナイル熱について簡便な診断法の開発を試みた。ハンタウイルスの核蛋白質抗原とし、様々なげっ歯類の抗体検出が可能なELISAとイムノクロマトグラフィー法を開発した。IgGもしくはStrep-tagを付加したダニ媒介性脳炎ウイルスのウイルス様粒子 (SPs) を用いた抗体検出用のELISAを開発した。相同組換えを利用した、安定性の高いウエストナイルウイルスのリバースジェネティック法を開発した。
  • 近隣地域からの侵入が危惧されるわが国にない感染症の発生予防に関する研究
    厚生労働省:厚生労働科学研究費補助金
    研究期間 : 2013年04月 -2016年03月 
    代表者 : 苅和 宏明
  • 日本学術振興会:科学研究費助成事業 若手研究(B)
    研究期間 : 2012年04月 -2014年03月 
    代表者 : 好井 健太朗, 苅和 宏明
     
    フラビウイルスには人獣共通感染症の原因ウイルスが多く属しており、その中にはヒトや家畜に重篤な脳炎を引き起こす病原体もある。本研究では、フラビウイルス特異的抵抗性遺伝子であるOas1bを正常に発現するコンジェニックマウスを使用することで、脳炎のより詳細な病態解析が可能なことを示した。さらに、ダニ媒介性フラビウイルスの高病原性化には、哺乳動物への適応により生じる3'非翻訳領域の変異が重要であることを明らかにした。
  • 海外からの侵入が危惧される野生鳥獣媒介性感染症の疫学、診断・予防法等に関する研究
    厚生労働省:厚生労働科学研究費補助金
    研究期間 : 2010年04月 -2013年03月 
    代表者 : 苅和 宏明
  • わが国とアメリカ大陸のウイルス性人獣共通感染症の流行阻止のための研究
    日本学術振興会:科学研究費補助金
    研究期間 : 2009年04月 -2013年03月 
    代表者 : 苅和 宏明
  • 日本学術振興会:科学研究費助成事業 基盤研究(B)
    研究期間 : 2009年 -2012年 
    代表者 : 苅和 宏明, 有川 二郎, 好井 健太朗, 森松 組子, 高島 郁夫
     
    メキシコのげっ歯類から3種類の新規ハンタウイルスを検出した。各種ハンタウイルス感染の検査に応用可能な抗体検出法を開発した。メキシコのハンタウイルスのヌクレオキャプシド(NP)に対して作出されたモノクローナル抗体6種類のうち5種類はげっ歯類を宿主とするハンタウイルスに対して広く反応した。ウエストナイルウイルスのE蛋白上の糖鎖の有無がニワトリのヒナにおける病原性発現に大きな影響を与えることが判明した。
  • 日本学術振興会:科学研究費助成事業 若手研究(B)
    研究期間 : 2010年 -2011年 
    代表者 : 好井 健太朗, 高島 郁夫, 苅和 宏明, 安居院 高志
     
    感染症に対する宿主側の自然免疫による防御機構において、オリゴアデニル酸合成酵素(OAS)は重要な働きを持つ。中でもげっ歯類のOAS遺伝子ファミリーのOas1bはフラビウイルス感染を特異的に防御する。本研究では、このOas1bによるフラビウイルス特異的な感染防御機構を解析することで、OASがフラビウイルスの感染時におけるウイルスの排除や病態形成に重要な役割を果たしていることを示した。
  • 国内で発生のないベクター媒介性感染症の疫学診断法等の研究
    厚生労働省:厚生労働科学研究費補助金
    研究期間 : 2007年04月 -2010年03月 
    代表者 : 苅和 宏明
  • 日本に侵入する危険性の高い北米産ウイルス性人獣共通感染症の疫学的研究
    日本学術振興会:科学研究費補助金
    研究期間 : 2004年04月 -2008年03月 
    代表者 : 苅和 宏明
  • 日本学術振興会:科学研究費助成事業 基盤研究(B)
    研究期間 : 2006年 -2008年 
    代表者 : 有川 二郎, 森松 組子, 遠藤 理香, 苅和 宏明, 森松 組子, 苅和 宏明, 垂石 みどり
     
    ハンタウイルスは持続感染齧歯類が感染源となる人獣共通感染症でヒトに腎症候性出血熱やハンタウイルス肺症候群という重篤な疾病を引き起こす。齧歯類は血中に高い中和抗体を保有しつつ不顕性にウイルスを排泄し続けるというきわめて特徴的な持続感染を成立させている。本研究では齧歯類におけるウイルス特異的CTL抑制を中心とする免疫抑制を介したハンタウイルス持続感染成立機構の詳細を明らかにすることを目的とする。
  • 日本学術振興会:科学研究費助成事業 基盤研究(A)
    研究期間 : 2005年 -2008年 
    代表者 : 高島 郁夫, 苅和 宏明, 前田 秋彦, 有川 二郎, 橋本 信夫
     
    近年、国内外で問題となっているウイルス性人獣共通感染症のうち、侵入または流行の危険性があり、重篤に経過し致死率が高いウエストナイル熱、ダニ媒介性脳炎およびハンタウイルス感染症について診断法を開発し、疫学調査を実施し、分離ウイルスの性状解析を行った。 ウエストナイルウイルスと日本脳炎ウイルスの遺伝子鑑別診断法としてリアルタイムPCR法を開発した。さらに準ウイルス粒子とウイルス様粒子を用いた血清学的な鑑別診断法を開発した。極東ロシアの野鳥の中和抗体を測定したところ、91羽中15羽(16.5%)にウエストナイル特異的な中和抗体が検出されたことから、極東ロシアの野鳥間にウエストナイルウイルスの流行していることが示唆された。 ダニ媒介性脳炎については準ウイルス粒子を用いたヒトおよび野ネズミ用の抗体検出ELISA を開発した。本ELISAを用いて国内各地の野ネズミ血清について抗体調査を実施したところ、島根県の野ネズミ58検体中2検体がダニ媒介性脳炎ウイルス特異抗体陽性となり、新しいウイルス汚染地が特定された。 ハンタウイルス感染症では、ウイルス核タンパクを対象にしたヒトおよび野ネズミの抗体検出ELISA および抗原検出ELISAを開発した。この抗体検出ELISAを用いてタイのレプトスピラ感染症を疑われた患者に、ハンタウイルス抗体陽性例を検出したため、ハンタウイルス感染症と症状の関連が示唆された。ロシアのボルガ川流域のサマラ市において、野ネズミの疫学調査を行い、流行中のウイルスはPuumala型のハンタウイルスであることを明らかにした。北海道の中川町と当別町においてエゾヤチネズミの疫学調査を行い、ハンタウイルスの野ネズミ集団内では、オスがメスよりも有意に高い感染率を示すことを明らかにした。
  • 日本に侵入する危険性の高い北米産ウイルス性人獣共通感染症の疫学的研究
    日本学術振興会:科学研究費助成事業 基盤研究(B)
    研究期間 : 2004年 -2007年 
    代表者 : 苅和 宏明, 高島 郁夫, 有川 二郎, 吉松 組子
     
    本研究では、日本国内に侵入または流行の危険性の高いウエストナイル熱とハンタウイルス感染症について、診断法を開発するとともに、疫学調査を実施した。 1)ウエストナイル熱の診断法開発:ウエストナイルウイルス感染の日本への侵入を監視するため、抗ウエストナイルウイルス抗体の検出法として微量中和法や阻害ELISA法を開発した。 2)ロシア国内で捕獲された鳥類におけるウエストナイルウイルスの感染調査:ロシアウラジオストック近郊のハンカ湖とハバロフスク北部のアニュイ川河畔で捕獲された98例の鳥類の腎臓からReal-time PCRによりウエストナイルウイルスの遺伝子検出を行ったが、いずれの検体からもウイルス遺伝子は検出されなかった。しかし、微量中和法により血清の得られた91例中うち15例から抗ウエストナイルウイルス抗体が検出され、極東ロシアで野鳥においてウエストナイルウイルスの感染のあることが示唆された。極東ロシアからは日本に渡り鳥が多数飛来することから、今後わが国におけるウエストナイル熱の防疫体制の強化が強く望まれる。 3)メキシコのげっ歯類におけるハンタウイルス感染症の疫学調査:北アメリカ産ハンタウイルスのS遺伝子を標的としたRT-PCRによる遺伝子検出法を開発した。そこでメキシコ国内で捕獲されたげっ歯類213例の肺からRNAを抽出し、RT-PCRによりハンタウイルス遺伝子の検出を試みたところ、21例からウイルス遺伝子が検出された。これらの遺伝子の検出された個体の21例中19例では抗体も検出された。したがって、これまでハンタウイルスの流行が明らかにされていなかったメキシコにおいて、げっ歯類にハンタウイルスが感染していることが判明した。
  • 動的構造解析法によるプリオンの構造と異常型への変遷機構の解明
    日本学術振興会:科学研究費助成事業 基盤研究(B)
    研究期間 : 2005年 -2006年 
    代表者 : 稲波 修, 堀内 基広, 苅和 宏明, 稲葉 睦, 桑原 幹典
     
    BSEやCJDなどのプリオン病では、構造異常を呈する異常型プリオンタンパク質(PrPSc)が正常型PrPCと相互作用し、その高次構造を異常型に変換させ、病態を引き起こすと考えられている。このPrPの構造変化機構には、酸性エンドソームが関与していると考えられている。本研究ではsite-directed spin labeling (SDSL)法を用い、αヘリックス1(H1)の裏側、βシート1(S1)とβシート2(S2)の3つのpH感受性領域をESRによる解析で見出した。近年、molecular dynamics (MD) simulationを用いた研究により、酸性pHが引き起こすPrPSc様のβ-sheet-rich構造への転換にアスパラギン酸177(Dl77)とヒスチジン186(H186)が重要な役割を持つと報告されたが、実際には実験的な証明はなされていない。本研究ではさらにD177とR163が形成する塩橋とH176とH186の2つのヒスチジン残基について、β-sheet2(S2)のpH感受性に対する影響をSDSL-ESR法で解析した。その結果、D177が形成する塩橋の形成がS2の中性付近でのβシート構造の維持に重要であることが明らかとなったが、近傍のH176の存在がこの構造安定性にも重要であることが見いだされた。また、H186のCJD変異についてはS2の構造安定化には寄与しておらず、βシート領域以外の別の構造変化がPrps^cの形成の引き金となることが示唆された。以上の結果は、D178Nといったアミノ酸の突然変異で発症する遺伝性CJDやFFIは、S2の構造変化が引き金となりPrPScが形成されると考えられる。また、本研究では、シングルラベルしたプリオンタンパク質にパルスESR法を適応することにより、低pHあるいは塩酸グアニジンにより誘導されるプリオン凝集体で見られる距離情報を得ることが出来ることも見いだした。これは将来、他種類のシングルラベルしたプリオンタンパク質に適応することにより、異常型プリオンの構造情報を得ることが出来る方法として期待できるものである。
  • 日本と極東ロシアの野生げっ歯類を病原巣動物とする人獣共通感染症の比較疫学的研究
    日本学術振興会:科学研究費補助金
    研究期間 : 2001年04月 -2004年03月 
    代表者 : 苅和 宏明
  • わが国のげっ歯類を病原巣動物とするウイルス性人獣共通感染症の疫学的研究
    日本学術振興会:科学研究費補助金
    研究期間 : 2001年04月 -2004年03月 
    代表者 : 苅和 宏明
  • 西ナイルウイルスの日本上陸に備えた、蚊における新しい感染防御システムの開発
    日本学術振興会:科学研究費助成事業 基盤研究(C)
    研究期間 : 2003年 -2004年 
    代表者 : 水谷 哲也, 江下 優樹, 苅和 宏明, 倉根 一郎
     
    本研究の目的は、蚊の発育におけるJNKの生物学的機能を明らかにするとともに、JNKの機能を阻害することにより蚊の発育を抑制する方法を確立し、新しい殺虫剤の開発のための基礎研究を行なうことにあった。 平成15,16年度の研究から以下のことが明らかになった。 1.JNKの阻害剤(SP600125)をヒトスジシマカの1齢幼虫に暴露することにより、発育を阻害することに成功した。蚊の培養細胞では、JNKは抗アポトーシスの働きをしていることを確認した。 2.リポポリサッカライド(LPS)を蚊の細胞に添加することにより、JNKの活性化が観察され、抗細菌ペプチドをコードしていると考えられる新規遺伝子を発見した。 3.JNKの機能を幼虫で評価するために、siRNAによる機能阻害方法について検討した。3齢幼虫にsiRNAをマイクロインジェクションすることにより成虫になる割合を激減させることに成功したが、多くの幼虫に施すことは不可能であった。そこで、市販のトランスフェクション試薬を検討したが、マイクロインジェクションに勝る方法を発見することはできなかった。導入方法については引き続き検討を行なっている。 4.放線菌から採取された種々のキチナーゼの阻害剤を含むと思われる培養液中で1齢幼虫を飼育し、2種類の培養液で発育が阻害された。阻害物質をコードする遺伝子を同定している。 本研究から、ヒトスジシマカの幼虫においてJNKが重要な働きをしていることが明らかとなった。また、キチナーゼを阻害することも発育阻害に重要なことが明らかとなった。
  • 西ナイル熱ウイルスなどのフラビウイルス感染症の診断法、疫学および予防法
    日本学術振興会:科学研究費助成事業 基盤研究(A)
    研究期間 : 2002年 -2004年 
    代表者 : 高島 郁夫, 苅和 宏明, 森田 公一, 只野 昌之, 竹上 勉, 江下 優樹, 水谷 哲也
     
    本研究において、日本に存在しているかまたは侵入の可能性のあるフラビウイルス感染症の流行予防のための診断法の開発、病原性の解明および蚊の調査に関する研究を実施して以下の成果を得た。 1.診断法の開発 1)ウエストナイル熱について、RT-PCR RFLP法、リアルタイムPCR法およびRT-LAMP法による遣伝子診断法を開発した。この方法によると簡便な設備により、ウエストナイルウイルスと日本脳炎ウイルスおよびウエストナイルウイルス株間の簡別診断が可能となった。 2)ダニ媒介性脳炎の迅速な血清診断法として、ウイルス株粒子を用いたヒト用のIgGおよびIgM-ELISA法を開発した。 2.病原性の解明 1)ウエストナイルウイルスの神経侵襲性毒力がウイルスエンベロープ蛋白への糖鎖付加により決定されることを明らかにした。 2)ダニ媒介性脳炎ウイルスの神経侵襲性毒力は、ウイルスエンベロープ蛋白の1個のアミノ酸の置換により電荷が陽性に変化することにより低下することが示された。 3)ダニ媒介性脳炎ウイルスの感染性cDNAクローンを用いた解析により、NS5とエンベロープ蛋白における各々一ヶ所のアミノ酸変異が神経毒力の低下に相乗的に作用していることが示唆された。 3.蚊の調査 1)ウエストナイルウイルスに対する、日本産蚊4種アカイエカ、イナトミシオカ、ヒトスジシマカの感受性、およびアカイエカの媒介能を証明した。 2)ウエストナイルウイルス・デングウイルス媒介蚊ヒトスジシマカのJNKタンパクの機能を解析した。
  • 組換え可溶性外被蛋白を用いたハンタウイルスワクチンおよび迅速診断キットの開発
    日本学術振興会:科学研究費助成事業 基盤研究(B)
    研究期間 : 2001年 -2003年 
    代表者 : 森松 組子, 小野 恵利子, 森松 正美, 苅和 宏明, 有川 二郎
     
    1.CAG promotorを用いて発現させた組み換えハンタウイルスエンベロープ蛋白が細胞融合活性を持つことを明らかにした。ハンタウイルスの中和エピトープの中央に細胞融合関連エピトープが位置することから、この組み換え蛋白が中和エピトープを保持していることが示された。また、同時に核蛋白抗原もCAG promotorを用いて発現させることに成功し、エンベロープ蛋白との相互作用について検討した。その結果核蛋白はエンベロープ蛋白の発現と輸送を押さえる可能性があること。この二つの蛋白のみではウイルス様粒子は形成されないことが示された。 2.CAG promotorを用いて発現させた組み換えハンタウイルスエンベロープ蛋白を用いて、外皮のみがハンタウイルスエンベロープ蛋白であり、内部が不稔のvesicular stomatitis virus (VSV)であるPseudotype VSV (VSVΔG^*HTN)を作製し、迅速で安全な中和抗体測定システムを構築した。また、分泌型の組み換えハンタウイルスエンベロープ蛋白を抗原としてマウスへ免疫し、中和抗体の誘導の可否を検討した。その結果、中和抗体は誘導されなかった。そのため、次にVSVΔG^*HTN粒子を抗原としてワクチネーションを試みた。その結果、免疫されたマウスにはエンベロープ蛋白に対する抗体の上昇が確認された。さらにこのマウスに攻撃接種したところ、核蛋白に対する抗体の上昇、および活性はほとんど見られず、感染が防御されたと考えられた。以上のことから、エンベロープ蛋白をウイルス様にパッケージングすることで、感染防御活性を有する抗体応答が得られることが分かった。これらの結果から、この組み換え蛋白を用いた安全なワクチン開発の可能性が示された。
  • 日本と極東ロシアの野生げっ歯類を病原巣動物とする人獣共通感染症の比較疫学的研究
    日本学術振興会:科学研究費助成事業 基盤研究(B)
    研究期間 : 2001年 -2003年 
    代表者 : 苅和 宏明, 有川 二郎, 水谷 哲也, 高島 郁夫, 岩崎 琢也
     
    本研究ではげっ歯類を病原巣動物とする人獣共通感染症のうち、ハンタウイルス感染症とダニ媒介性脳炎を対象に主にげっ歯類を中心に疫学調査を行い、両感染症の極東ロシアにおける感染状況を明らかにすることを試みた。 これまでに本研究により極東ロシアのウラジオストック周辺で捕獲されたハントウアカネズミがヒトに重症型の腎症候性出血熱(HFRS)を引き起こす新型のハンタウイルス(Amurウイルス)を保有していることが判明した。ハントウアカネズミは中国や韓国、および北海道にも分布することから、東アジアでハントウアカネズミによって媒介されるHFRSが発生していることが示唆された。極東ロシアで検出されたAmurウイルスに近縁のウイルス(H5とB78株)が中国のHFRS患者から分離されていることから、これらの中国由来株の遺伝子性状と抗原性状について解析を行い、H5とB78株のSとM遺伝子全長の塩基配列を明らかにした。これら中国由来株と極東ロシアのAmurウイルスの核蛋白のアミノ酸配列はそれぞれ98%以上の高い一致率を示した。また中国株のウイルスの抗原性はこれまで知られていたHantaan型と明らかに異なることが判明した。ダニ媒介性脳炎ウイルスの北海道分離株であるOshima5-10株をプラック精製して、大型のプラックを形成する細胞馴化株を得た。本株の遺伝子解析の結果、ウイルス糖蛋白上の糖付加部位のアミ.ノ酸が変異して糖鎖が失われたことが明らかになった。さらにこの株はマウスに対して親株よりも非常に弱い病原性しか示さないことから、糖鎖の付加がウイルスの毒力に関連することが示された。
  • わが国のげっ歯類を病原巣動物とするウイルス性人獣共通感染症の疫学的研究
    日本学術振興会:科学研究費助成事業 基盤研究(C)
    研究期間 : 2001年 -2003年 
    代表者 : 苅和 宏明, 有川 二郎, 水谷 哲也, 高島 郁夫, 福島 博, 土屋 公幸
     
    本研究ではげっ歯類を病原巣動物とする人獣共通感染症のうち、ハンタウイルス感染症とダニ媒介性脳炎を対象に主にげっ歯類を中心に疫学調査を行い、両感染症の日本における感染状況を明らかにすることを試みた。 まず、北海道、宮崎県、富山県、島根県、でげっ歯類の捕獲調査を行い、捕獲されたげっ歯類についてハンタウイルス抗体の検出を試みた。北海道では捕獲されたげっ歯類のうち、,エゾヤチネズミとドブネズミで抗体が検出された。特に千歳空港で捕獲されたエゾヤチネズミと小樽港のドブネズミに抗体が検出されたことから、空港と港湾でのネズミ類の防除対策の強化が必要と考えられた。北海道のエゾヤチネズミが保有するウイルスを分離するために、様々な動物にウイルスを接種して最適な動物種を探索したところ、スナネズミが実験動物の中でも最も感受性が高かった。そこで、スナネズミにエゾヤチネズミの材料を接種して2回継代し、さらに培養細胞に接種したところ、接種細胞中にウイルス抗原とウイルス遺伝子が検出され、分離に成功したことが確認された。さらに本ウイルスのS遺伝子とM遺伝子の全長の塩基配列の決定により、それぞれの遺伝子がコードする核蛋白と糖蛋白のアミノ酸配列が明らかになった。本ウイルスの核蛋白と糖蛋白のアミノ酸配列は最も近縁と考えられていたヨーロッパのPuumalaのウイルスとそれぞれ5%と10%の相違があり、本ウイルスが独立したウイルス型に属するものと考えられた。 また、ダニ媒介性脳炎の抗体測定にはこれまで中和試験が主に使用されてきたが、危険度の高い生きたウイルスを用いたことや、時間がかかるなどの欠点があった。そこで、ウイルス遺伝子を含まない非感染性のウイルス様粒子(VLP)を作出し、VLPを用いたELISAを開発した。本法により、ダニ媒介性脳炎患者の抗体を安全で迅速に検出することが可能になった。
  • SARSコロナウイルスのウイルス粒子形成機構に関する研究
    研究期間 : 2003年
  • Study on the mechanism of virion formation of SARS-coronavirus
    研究期間 : 2003年
  • ダニ媒介脳炎の鑑別診断法とワクチンの開発に関する研究
    日本学術振興会:科学研究費助成事業 基盤研究(B)
    研究期間 : 2000年 -2002年 
    代表者 : 高島 郁夫, 高橋 健一, 水谷 哲也, 苅和 宏明, 倉田 毅, 根占 哲也
     
    本研究では北海道で近年発見されたダニ媒介脳炎(TBE)の具体的予防対策のために、安全で簡便な血清診断法を開発し、血清疫学調査によりウイルス汚染地を特定した。さらに近年ロシア等の流行地で流行しているウイルス株に対する市販のワクチンの効果を評価した。 まずTBEウイルス渡島株に対して単クローン性抗体(Mab)を作出しこれらMabを用いた蛍光抗体法により、迅速なウイルス同定法を確立した。次に1999年〜2000年にロシアのハバロフスク市周辺、ウラジオストック市周辺およびイルクーツク市周辺で採集したマダニ類からウイルスを分離して遺伝子解析を行った。ハバロフスクおよびウラジオストックから分離されたTBEウイルス株はすべて極東型と同定された。一方イルクーツクからのウイルス株はすべてシベリア型と同定された。TBEウイルス極東型株とシベリア型株に対するヨーロッパ型TBEウイルスワクチンの効果を判定した。ワクチン接種者は極東型およびシベリア型TBEウイルスに対して良好な中和抗体応答を示した。さらにマウスにおけるワクチン効果を判定したところ、ワクチン接種マウスは両ウイルス株の攻撃に対して有意な防御を示した。従ってTBEウイルスヨーロッパ型ワクチンは現在シベリアと極東地区に流行しているTBEウイルスに有効であることが判明した。遺伝子工学的手法により、TBEウイルスのウイルス様粒子(VLP)を作出し、ELISAによるTBEの血清診断に応用した。IgM-ELISAにより極東ロシアのTBE患者血清を用いて調べたところ、敏感度と特異性に優れた方法であることが判明した。
  • 日本で分離されたダニ媒介脳炎ウイルスの病原性と起源に関する研究
    日本学術振興会:科学研究費助成事業 基盤研究(B)
    研究期間 : 1999年 -2001年 
    代表者 : 高島 郁夫, 高橋 健一, 岩崎 琢也, 苅和 宏明, 水谷 哲也, 丸山 務
     
    本研究ではダニ媒介脳炎の予防対策の資料とするため、北海道におけるウイルス汚染地の特定、マウスを用いた北海道株の病原性の評価およびヨーロッパ型ワクチンの北海道ウイルス株に対する効果判定について検討を加えた。さらに北海道株の分岐時代を極東型との系統学的な比較により推定した。またシベリア地域に分布するウイルス株の亜型を明らかにした。 ダニ媒介性脳炎(TBE)ウイルス北海道Oshima株はマウスにおいて他のTBEウイルス株と共通の病原性を示した。北海道におけるTBEウイルスの汚染地はウマとイヌの血清疫学調査によって渡島、檜山、胆振および後志の道南4支庁に及ぶことが判明した。エゾアカネズミとエゾヤチネズミからTBEウイルスが分離されたことから、これらの野ネズミがTBEウイルスの病原性動物と推定された。北海道と極東ロシアで分離されたTBEウイルスの系統樹解析を実施した。北海道株は極東型と判明した。さらに同義置換率の解析からこれらのウイルスの進化速度を算出し、分岐年代を推定したところ、北海道と極東株は260〜430年前に分岐したと推定された。シベリアと極東ロシアに分布するTBEウイルス株の性状を解析した。この結果極東型ウイルスに加え新しいシベリア型ウイルスの存在が明かとなった。ヨーロッパ型のTBEウイルスワクチンの効果をマウスでの防禦試験とヒトでの中和抗体産生能で調べた。本ワクチンはTBEウイルス北海道株、極東株およびシベリア株に有効であることが判明した。
  • 実験用小動物から微量部分採血された血清を用いた感染症血清診断法の確立に関する研究
    日本学術振興会:科学研究費助成事業 基盤研究(B)
    研究期間 : 1999年 -2001年 
    代表者 : 有川 二郎, 森松 組子(吉松 組子), 苅和 宏明, 佐藤 浩, 高倉 彰
     
    1.ハンタウイルス抗体測定用の抗原として、大腸菌とバキュロウイルスを発現ベクターとしてハンターン型、ソウル型およびドブラバ型のそれぞれの核蛋白抗原の発現に成功し、いずれも、ELISA法診断用抗原として利用が可能である活性と量が得られることが判明した。 2.バキュロウイルス発現ベクターによって、核蛋白のN末端50アミノ酸を削除した抗原を発現させてELISAへ応用すると、血清型鑑別抗原として応用可能であることが、患者血清および感染げっ歯類血清を用いて確認された。 3.マウス肝炎ウイルス(MHV)核蛋白コード遺伝子をイーストの系で発現することに成功した。培養上清中の発現のピークは培養後72時間であった。しかし、ELISAにおける反応性が低く、発現系および核蛋白内の発現部位の再検討が今後の課題である。 4.リンパ球性脈絡髄膜炎(LCM)ウイルス国内分離株及びプロトタイプ株の核蛋白コード領域遺伝子をクローニングし、全長、C末領域および中間領域の3つの領域についてそれぞれCOS細胞での発現に成功した。それぞれの両域抗原の診断的有用性の検討が今後必要である。 5.LCMウイルス核蛋白発現バキュロウイルスを国立感染症研究所から分与を受け、その抗原性を検討した。その結果、感染Sf9細胞はIFA抗原として、また、Tm5細胞にて増殖させたウイルスはELISA抗原に適していることが判明した。本抗原と両診断法を用いて、わが国のマウス動物実験施設、延べ1,117施設由来9,840検体を対象に血清のスクリーニングを行った結果、ELISAは大量検体処理のためのスクリーニング試験に、IFAはその結果の確認試験に有用であることが示された。
  • 本邦と極東ロシアのげっ歯類におけるハンタウイルスの比較疫学的研究
    日本学術振興会:科学研究費補助金
    研究期間 : 1998年04月 -2000年03月 
    代表者 : 苅和 宏明
  • ハンタウイルスの病原性の分子生物学的解析
    日本学術振興会:科学研究費助成事業 基盤研究(B)
    研究期間 : 1999年 -2000年 
    代表者 : 有川 二郎, 森松 組子, 苅和 宏明, 高島 郁夫, 森川 茂
     
    1.中国側研究分担者の所属研究機関を訪問し中国では地域的に強毒と弱毒のハンタウイルスが流行している可能性が示唆された。また、中国におけるワクチン開発の現状と有効性について情報を得た。さらに各国流行株と中国株の遺伝子の相互関係の解析ならびに血清型鑑別診断法の技術交流を行った。 2.スロバキア研究分担者の研究所を訪問し、欧州におけるハンタウイルス遺伝子解析に関する情報交換と技術習得を行った。 3.日本側研究協力者を米国側研究分担者の研究所に派遣しレセプター解析と遺伝子再集合ウイルス作製のための情報交換と技術交流を行った。 4.日本側研究協力者を米国Wisconsin大学獣医学部に派遣しcDNAから人工的にハンタウイルスを合成するための基礎的技術の習得を行った。 5.ヘルシンキ大学、Vaheri博士から北欧流行ハンタウイルスの病原性規定因子についての研究打ち合わせを行うと同時に、当該研究分野の他の研究者と共に情報交換を行った。 6.東南アジア流行ウイルスとの比較を目的にウイルス株および患者と感染動物の血清の取得の可能性について情報交換を行った。 7.ハンタウイルスの感染がGalNacを特異的に認識するレクチンであるDBAとSBAによって増強されること、ならびに感染過程でのウイルスと細胞膜の融合には細胞側の因子が関与することを明らかにした。 8.マウスへの病原性が末梢組織での増殖効率が高いことが関連し、エンベロープ蛋白の単一アミノ酸の変異が規定していることを明らかにした。 9.遺伝子再集合ウイルス作成のためにウイルス遺伝子の発現をMとS遺伝子について完了した。また、転写、複製機構解明のためのミニゲノムを作成した。今後、それら遺伝子発現蛋白とミニゲノムの発現の確認が課題である。
  • ハンタウイルスの中枢神経系及び免疫系組織における持続感染成立機構解析に関する研究
    日本学術振興会:科学研究費助成事業 基盤研究(A)
    研究期間 : 1998年 -2000年 
    代表者 : 有川 二郎, 森川 茂, 苅和 宏明, 森松 組子
     
    1.ハンタウイルスの感染効率が、N-アセチルガラクトサミン結合性レクチン(DBAとSBA)を添加することにより約10倍増強され、細胞表面上のレセプター又はその近傍にあるN-アセチルガラクトサミンとウイルスをレクチンが架橋することにより吸着効率が増強するためと考えられた。 2.ハンタウイルス感染後、細胞融合を惹起する細胞株と起こさない細胞株の確立に成功し、ハンタウイルスエンベロープ蛋白が引き起こす感染細胞融合には細胞側因子も関与していることを明らかにした。 3.強毒株感染マウスでは脳へのウイルスの到達が弱毒株に比べ4-5日早いことがその後の全身感染の拡大と致死的転帰につながるものと考えられた。 4.強毒株は脳微小管内皮細胞と腹腔マクロファージにおいて弱毒株よりも高い増殖率を示したが、神経芽腫細胞では両ウイルスに差は認められなかったことからウイルスの病原性には末梢の標的細胞での増殖率が関連していると考えられた。 5.強毒と弱毒ハンタンウイルス間で遺伝子分節の交換された遺伝子再集合ウイルスを作成し、病原性と関連するウイルス側の要因を解析した結果、Mセグメント遺伝子中のアミノ酸変異(515番目アミノ酸、IleからThr)が病原性に関連し、Lセグメントも部分的に関連することが明らかになった。 6.強毒株感染SCIDマウスに、感染3週後のBALB/cマウス由来脾臓細胞を移入したところ、感染3週目に移入した場合、移入4日目から著しい体重減少が一過性に認められ、免疫関連の病原性発現機構が確認された。 7.ハンタンウイルスの核蛋白遺伝子(S分節)とエンベロープ蛋白遺伝子(M分節)をクローニングし、哺乳動物細胞(293T細胞)で高率に発現させることに成功した。 8.Lセグメントの発現系の確立を目的にLセグメントRNA全長のクローニングに成功した。今後、この発現の有無について検討を加える予定である。
  • 本邦と極東ロシアのげっ歯類におけるハンタウイルスの比較疫学的研究
    日本学術振興会:科学研究費助成事業 基盤研究(C)
    研究期間 : 1998年 -1999年 
    代表者 : 苅和 宏明, 水谷 哲也, 有川 二郎, 高島 郁夫, 吉松 組子
     
    我が国や極東口シアにおいてハンタウイルスの疫学調査を行い、以下のことが明らかになった。 1.実験的に作出した感染ラットの尿中にウイルスが排泄され、この尿を感受性ラットに経鼻的に滴下することにより感染が成立すること、さらに尿接種によって感染したラットの尿からもウイルスが検出されたことから、尿を介した経鼻伝播で本ウイルスが自然界で維持されうることが明らかになった。 2.パキュロウイルスで様々なハンタウイルスの核蛋白を発現させ、これらを抗原に用いたELISA法を開発した。本法により、抗体保有が判定できるばかりでなく、血清を異なった抗原に反応させることで、感染したハンタウイルスの型までも類推することが可能となった。 3.ハンタウイルス肺症候群(HPS)患者の体内のウイルス量を測定するために定量的PCR法を開発した。このPCR法により、HPSの急性期には高力価のウイルスが血中に存在するが、感染の回復期にはウイルスはもはや血中から消失することが明らかにされた。 4.複数の血清診断法を用いて、我が国のヒトにおける大規模なハンタウイルスの血清疫学調査を行った。健常者では抗体保有者は全く存在しないにもかかわらず、原因不明の肝炎患者においては約2〜3%の患者がハンタウイルス抗体を保有することが判明した。これにより、本邦でも一般の市民にハンタウイルスの感染があることが明らかになった。 5.極東口シアのウラジオストックでげっ歯類の捕獲調査を行い、日本の北海道と同じくエゾヤチネズミがハンタウイルスに感染していることが明らかになった。また、日本には生息していないセスジネズミも抗体を保有しており、極東ロシアにおいて強毒のHantaan型のハンタウイルスの存在が示唆された。また、ヨシハタネズミからウイルス遺伝子が検出され、塩基配列の解読を行ったところ、まったく新しい型のハンタウイルスであることが、判明した。
  • フラビウイルスの疫学的研究
    研究期間 : 1999年
  • Epidemiological study of flaviviruses
    研究期間 : 1999年
  • 野生げっ歯類を病原巣とする人畜共通伝染病の診断法、予防法および疫学に関する研究
    日本学術振興会:科学研究費助成事業 基盤研究(A)
    研究期間 : 1996年 -1998年 
    代表者 : 高島 郁夫, 丸山 務, 森田 千春, 金子 賢一, 有川 二郎, 苅和 宏明, 小川 益男
     
    1. ダニ媒介脳炎の成績:ウマ、イヌおよび野ネズミの血清を用いた疫学調査を実施したところ、ウイルスの汚染地は道南の4支庁を中心に広範に分布していることを明らかにした。ダニ媒介脳炎の患者発生地区で採集したヤマトマダニ600匹から2株のウイルス分離に成功した。さらに、エゾアカネズミ11匹中1匹とエゾヤチネズミ79匹中1匹からダニ媒介性脳炎ウイルスを分離した。先に犬から分離したダニ媒介脳炎ウイルスのOshima株の病原性をマウスモデルで検討したところ、本ウイルス株はダニ媒介脳炎ウイルスに共通する神経侵襲性を示した。オーストリア産のダニ媒介脳炎ウイルスワクチンはOshima株に有効なことがマウスの実験感染とヒトの中和抗体産生で証明された。 2. ハンタウイルス感染症の成績:遺伝子学的手法を用いたハンタウイルスの血清診断法を確立するために、バキュロウイルス発現ハンタウイルス組み換え蛋白を用いたELISA法を開発し、実験用ラットを対象としたスクリーニングテストとして実用化した。 3. Q熱の成績:Q熱リケッチア(Coxiella burnetii)の日本分離株は、16SrRNAの塩基配列が,株間で99.4%と高い相同性を示し、これらは1属1種であることが確認された。さらに野外にはモルモットに対して病原性の異なるI相菌が存在することが明らかになった。 4. その他の人畜共通伝染病の成績:エルシニア属菌、サルモネラ属菌、紅班熱リケッチアに野ネズミが高率に汚染していることが明らかになった。
  • わが国の野性げっ歯類におけるハンタウイルスの感染調査と新型ハンタウイルスの分離
    日本学術振興会:科学研究費補助金
    研究期間 : 1995年04月 -1996年03月 
    代表者 : 苅和 宏明
  • ハンタウイルスの生態学的研究
    研究期間 : 1990年
  • Ecological study of hantaviruses
    研究期間 : 1990年


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