研究活動情報

• Lipid nanoparticle-encapsulated microRNA-192: An anti-inflammatory adjuvant that enhances vaccine efficacy in aged mice.
  Yuriko Takagi; Tasuku Nishimura; Suraiya Aktar; Daisuke Katayama; Ken Takashima; Tomomi Kawakita; Toshiki Sekiya; Masashi Shingai; Hiroki Tanaka; Hidetaka Akita; Yusuke Miyashita; Kimitoshi Nakamura; Takahisa Kouwaki; Hiroyuki Oshiumi
  Molecular therapy. Nucleic acids, 37, 1, 102784, 102784, 2026年03月12日, ［国際誌］
  英語, 研究論文（学術雑誌）, Proinflammatory cytokines are essential for initiating immune responses; however, excessive or aging-related chronic inflammation impairs immunity and reduces vaccine efficacy. In this study, we developed lipid nanoparticles (LNPs) encapsulating anti-inflammatory microRNA-192 (miR-192) to attenuate inflammation and improve vaccine performance in the elderly. Results revealed that specific proinflammatory cytokines, including interleukin (IL)-6 and tumor necrosis factor (TNF)-α at the vaccination site, diminished antigen-specific antibody production. Notably, miR-192 endowed LNPs with strong anti-inflammatory properties, markedly enhancing vaccine efficacy, especially in aged mice. Transcriptomic analyses demonstrated that miR-192 downregulated multiple pro-inflammatory cytokines, such as senescence-associated secreted phenotype factors, which hinder vaccine responses. Additionally, miR-192 inhibited key components of the JAK-STAT signaling pathway, crucial for cytokine receptor signaling in myeloid cells. Overall, these findings indicate that miR-192 effectively suppresses harmful inflammatory responses, substantially enhancing vaccine efficacy, and highlight the therapeutic potential of the anti-inflammatory microRNA-based adjuvants for improving vaccination outcomes in the elderly.
• A Case of Infection with Leptospires from Three Different Serovars During a Flood in the Philippines.
  Yasutake Yanagihara; Toshiyuki Masuzawa; Masashi Shingai; Marumi Ohno; Toshiki Sekiya; Naoki Nomura; Tomomi Kawakita; Chimuka Handabile; Yuichi Koshiishi; Richard Obeng-Kyeremeh; Toshihiro Ito; Mitsumasa Saito; Sharon Y A M Villanueva; Nina G Gloriani; Hiroshi Kida
  The American journal of tropical medicine and hygiene, 2025年07月01日, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, Leptospirosis a significant and life-threatening zoonosis with global reach. If diagnosis and treatment are delayed, the infection may lead to fatal Weil's disease. In the Philippines, a 21-year-old man was admitted to the hospital with leptospirosis-like symptoms, including fever, myalgia, headache, anuria, jaundice, hemorrhage, skin rash, and diarrhea. Although he was immediately treated with penicillin G, the patient died shortly after admission. The serological test result was negative for Leptospira; however, three leptospires were isolated from the patient through selective cultivation under varying conditions. Microscopic agglutination tests and a phylogenetic analysis of the flaB gene encoding the flagellin subunit protein revealed that the isolates belonged to three different serovars, suggesting that the patient was simultaneously infected with at least three distinct Leptospira serovars. In this case report, we highlight the potential risk of multiple infections with leptospires in humans, a critical consideration for diagnosis and treatment strategies.
• ARNAX, but not conventional adjuvants, alum and squalene, induces antigen-specific CD8+ T cell proliferation in vaccination with influenza split vaccine.
  Akari Yamaya; Kento Sonoda; Tomomi Kawakita; Yuichi Koshiishi; Masashi Shingai; Misako Matsumoto; Tsukasa Seya
  International immunopharmacology, 159, 114895, 114895, 2025年06月26日, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, The influenza split vaccine (ISV) is generally administered without adjuvant via intra-muscular (i.m.) or intranasal route. Research has demonstrated that SV alone exhibits no capacity to elicit a Th1 (T helper-1) response and cellular immunity represented by cytotoxic T lymphocytes (CTL) induction. This finding is consistent with the observation that SV merely up-regulates antibody (Ab) production in the pre-disposed population. This tendency is reproduced in the SV model in conjunction with conventional adjuvants, including alum or squalene. These adjuvants amplify the Th2 shift, consequently stimulating Th2-dependent Ab production in mice. Conventional adjuvants, when employed with SV in animal studies, demonstrate a limited capacity to stimulate cellular immune activation. The present study aims to demonstrate the efficacy of a Th1-inducing adjuvant, ARNAX, in promoting antigen-presenting DC and subsequent Ag-specific CTL proliferation, culminating in Th1 polarization. Consequently, the synthesis of the two effectors, CTL and Ab, is augmented by the incorporation of ARNAX in SV. The enhanced SV-mediated protective immunity may be further augmented by the induction of antigen-specific CTL and the unique profile of Th1-mediated Ab production.
• Optimization of the preparation method of inactivated intact virus particle vaccine for COVID-19.
  Marumi Ohno; Toshiki Sekiya; Richard Obeng-Kyeremeh; Chimuka Handabile; Minori Haruta; Naoki Nomura; Tomomi Kawakita; Masashi Shingai; Hiroshi Kida
  Vaccine, 56, 127173, 127173, 2025年05月22日, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, It has been recognized that it is difficult to maintain the virus particle structure of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which may be associated with lower immunogenicity of inactivated vaccines against coronavirus disease 2019 (COVID-19). We have previously demonstrated that an intact structure of the virus particles is critical for influenza inactivated whole virus particle vaccine (WPV) to be immunogenically potent. Here, we tested 37, 35, 33, and 31 °C for the virus propagation temperatures and the timing of formaldehyde treatment of the virus before and after centrifugation-based purification to obtain virus particles with an intact structure. Virus particles cultured at 33 °C retained spike proteins on the surface the most abundantly. The pretreatment of the virus with formaldehyde prevented the dissociation of the spike proteins from the viral surface during the centrifugation-based purification. The immunogenicity of the prepared vaccines, intact WPV and non-intact WPV that had lost the spike proteins, was evaluated in a mouse model. A single dose of intact WPV effectively induced humoral immunity compared to non-intact WPV, as indicated by higher titers of neutralizing antibodies. After a virus challenge, the mice vaccinated with a single dose of inactivated intact WPV showed less severe weight loss and lower virus titers in the lungs compared to those vaccinated with non-intact WPV. These results demonstrate the importance of the structural integrity of WPV in inducing effective and protective immunity, and provide significant insight into the development of COVID-19 WPV for practical use.
• ARNAX is an ideal adjuvant for COVID-19 vaccines to enhance antigen-specific CD4+ and CD8+ T-cell responses and neutralizing antibody induction.
  Tomomi Kawakita; Toshiki Sekiya; Yayoi Kameda; Naoki Nomura; Marumi Ohno; Chimuka Handabile; Akari Yamaya; Hideo Fukuhara; Yuki Anraku; Shunsuke Kita; Shinsuke Toba; Hirotake Tsukamoto; Tomohiro Sawa; Hiroyuki Oshiumi; Yasushi Itoh; Katsumi Maenaka; Akihiko Sato; Hirofumi Sawa; Yasuhiko Suzuki; Lorena E Brown; David C Jackson; Hiroshi Kida; Misako Matsumoto; Tsukasa Seya; Masashi Shingai
  Journal of virology, e0229024, 2025年04月15日, ［査読有り］, ［最終著者, 責任著者］, ［国際誌］
  英語, 研究論文（学術雑誌）, UNLABELLED: ARNAX is a synthetic nucleotide-based Toll-like receptor 3 (TLR3) ligand that specifically stimulates the TLR3/TIR domain-containing adaptor molecule 1 (TICAM-1) pathway without activating inflammatory responses. ARNAX activates cellular immunity via cross-presentation; hence, its practical application has been demonstrated in cancer immunotherapy. Given the importance of cellular immunity in virus infections, ARNAX is expected to be a more effective vaccine adjuvant for virus infections than alum, an adjuvant approved for human use that mainly enhances humoral immunity. In the present study, the trimeric recombinant spike protein of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was prepared as a vaccine antigen and formulated with ARNAX. When T-cell and neutralizing antibody responses were evaluated in immunized mice, antigen formulated with ARNAX generated significantly larger numbers of antigenspecific CD4+ and CD8+ T cells, as well as higher titers of neutralizing antibodies, compared to antigen alone or antigen formulated with alum. In experiments where immunized mice were challenged with a SARS-CoV-2 mouse-adapted virus derived from the ancestral strain, immunization with antigen formulated with ARNAX reduced virus titers in the lungs at 3 days post-infection to a much greater extent than did immunization with either antigen alone or that formulated with alum. These results show that ARNAX potently enhances the levels of both cellular and humoral immunity above those seen with alum, providing significantly greater viral clearing responses. Thus, ARNAX may act as a useful adjuvant for prophylactic vaccines, particularly for viral infectious diseases. IMPORTANCE: Cellular immunity is a critical immunological defense system against virus infections. However, aluminum salts, the most widely used adjuvant for vaccines for human use, do not promote strong cellular immunity. To prepare for the next pandemic of viral origin, the development of Th1-type adjuvants with low adverse reactions that induce cellular immunity is necessary. ARNAX is a TLR3 agonist consisting of DNA-RNA hybrid nucleic acid, which is expected to be an adjuvant that induces cellular immunity. The present study using a coronavirus disease 2019 mouse model demonstrated that ARNAX potently induces cellular immunity in addition to humoral immunity with minimal induction of inflammatory cytokines. Therefore, ARNAX has the potential to be used as a potent and welltolerated adjuvant for vaccines against pandemic viruses emerging in the future.
• Increased expression of CD38 on endothelial cells in SARS-CoV-2 infection in cynomolgus macaque
  Cong Thanh Nguyen; Misako Nakayama; Hirohito Ishigaki; Yoshinori Kitagawa; Akemi Kakino; Marumi Ohno; Masashi Shingai; Yasuhiko Suzuki; Tatsuya Sawamura; Hiroshi Kida; Yasushi Itoh
  Virology, 110052, 110052, Elsevier BV, 2024年03月, ［査読有り］
  研究論文（学術雑誌）
• Immunogenicity and protective efficacy of a co-formulated two-in-one inactivated whole virus particle COVID-19/influenza vaccine.
  Chimuka Handabile; Marumi Ohno; Toshiki Sekiya; Naoki Nomura; Tomomi Kawakita; Mamiko Kawahara; Masafumi Endo; Tomohiro Nishimura; Minako Okumura; Shinsuke Toba; Michihito Sasaki; Yasuko Orba; Brendon Y Chua; Louise C Rowntree; Thi H O Nguyen; Masashi Shingai; Akihiko Sato; Hirofumi Sawa; Kazumasa Ogasawara; Katherine Kedzierska; Hiroshi Kida
  Scientific reports, 14, 1, 4204, 4204, 2024年02月20日, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, Due to the synchronous circulation of seasonal influenza viruses and severe acute respiratory coronavirus 2 (SARS-CoV-2) which causes coronavirus disease 2019 (COVID-19), there is need for routine vaccination for both COVID-19 and influenza to reduce disease severity. Here, we prepared individual WPVs composed of formalin-inactivated SARS-CoV-2 WK 521 (Ancestral strain; Co WPV) or influenza virus [A/California/07/2009 (X-179A) (H1N1) pdm; Flu WPV] to produce a two-in-one Co/Flu WPV. Serum analysis from vaccinated mice revealed that a single dose of Co/Flu WPV induced antigen-specific neutralizing antibodies against both viruses, similar to those induced by either type of WPV alone. Following infection with either virus, mice vaccinated with Co/Flu WPV showed no weight loss, reduced pneumonia and viral titers in the lung, and lower gene expression of proinflammatory cytokines, as observed with individual WPV-vaccinated. Furthermore, a pentavalent vaccine (Co/qFlu WPV) comprising of Co WPV and quadrivalent influenza vaccine (qFlu WPV) was immunogenic and protected animals from severe COVID-19. These results suggest that a single dose of the two-in-one WPV provides efficient protection against SARS-CoV-2 and influenza virus infections with no evidence of vaccine interference in mice. We propose that concomitant vaccination with the two-in-one WPV can be useful for controlling both diseases.
• Extraction of the CDRH3 sequence of the mouse antibody repertoire selected upon influenza virus infection by subtraction of the background antibody repertoire
  Masashi Shingai; Sayaka Iida; Naoko Kawai; Mamiko Kawahara; Toshiki Sekiya; Marumi Ohno; Naoki Nomura; Chimuka Handabile; Tomomi Kawakita; Ryosuke Omori; Junya Yamagishi; Kaori Sano; Akira Ainai; Tadaki Suzuki; Kazuo Ohnishi; Kimihito Ito; Hiroshi Kida
  Journal of Virology, American Society for Microbiology, 2024年02月07日, ［査読有り］, ［筆頭著者］
  研究論文（学術雑誌）, As specific interactions between antigens and cell-surface antibodies trigger the proliferation of B-cell clones, the frequency of each antibody sequence in the samples reflects the size of each clonal population. Nevertheless, it is extremely difficult to extract antigen-specific antibody sequences from the comprehensive bulk antibody sequences obtained from blood samples due to repertoire bias influenced by exposure to dietary antigens and other infectious agents. This issue can be addressed by subtracting the background noise from the post-immunization or post-infection repertoire data. In the present study, we propose a method to quantify repertoire data from comprehensive repertoire data. This method allowed subtraction of the background repertoire, resulting in more accurate extraction of expanded antibody repertoires upon influenza virus infection. This accurate extraction of antigen- or infection-specific repertoire information is a useful tool for vaccine evaluation.
• The elucidation of plasma lipidome profiles during severe influenza in a mouse model.
  Marumi Ohno; Siddabasave Gowda B Gowda; Toshiki Sekiya; Naoki Nomura; Masashi Shingai; Shu-Ping Hui; Hiroshi Kida
  Scientific reports, 13, 1, 14210, 14210, 2023年08月30日, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, Although influenza virus infection has been shown to affect lipid metabolism, details remain unknown. Therefore, we elucidated the kinetic lipid profiles of mice infected with different doses of influenza virus A/Puerto Rico/8/34 (H1N1) (PR8) by measuring multiple lipid molecular species using untargeted lipidomic analysis. C57BL/6 male mice were intranasally infected with PR8 virus at 50 or 500 plaque-forming units to cause sublethal or lethal influenza, respectively. Plasma and tissue samples were collected at 1, 3, and 6 days post-infection (dpi), and comprehensive lipidomic analysis was performed using high-performance liquid chromatography-linear trap quadrupole-Orbitrap mass spectrometry, as well as gene expression analyses. The most prominent feature of the lipid profile in lethally infected mice was the elevated plasma concentrations of phosphatidylethanolamines (PEs) containing polyunsaturated fatty acid (PUFA) at 3 dpi. Furthermore, the facilitation of PUFA-containing phospholipid production in the lungs, but not in the liver, was suggested by gene expression and lipidomic analysis of tissue samples. Given the increased plasma or serum levels of PUFA-containing PEs in patients with other viral infections, especially in severe cases, the elevation of these phospholipids in circulation could be a biomarker of infection and the severity of infectious diseases.
• Two Modes of Th1 Polarization Induced by Dendritic-Cell-Priming Adjuvant in Vaccination.
  Tsukasa Seya; Masashi Shingai; Tomomi Kawakita; Misako Matsumoto
  Cells, 12, 11, 2023年05月29日, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, Viral infections are usually accompanied by systemic cytokinemia. Vaccines need not necessarily mimic infection by inducing cytokinemia, but must induce antiviral-acquired immunity. Virus-derived nucleic acids are potential immune-enhancers and particularly good candidates as adjuvants in vaccines in mouse models. The most important nucleic-acid-sensing process involves the dendritic cell (DC) Toll-like receptor (TLR), which participates in the pattern recognition of foreign DNA/RNA structures. Human CD141+ DCs preferentially express TLR3 in endosomes and recognize double-stranded RNA. Antigen cross-presentation occurs preferentially in this subset of DCs (cDCs) via the TLR3-TICAM-1-IRF3 axis. Another subset, plasmacytoid DCs (pDCs), specifically expresses TLR7/9 in endosomes. They then recruit the MyD88 adaptor, and potently induce type I interferon (IFN-I) and proinflammatory cytokines to eliminate the virus. Notably, this inflammation leads to the secondary activation of antigen-presenting cDCs. Hence, the activation of cDCs via nucleic acids involves two modes: (i) with bystander effect of inflammation and (ii) without inflammation. In either case, the acquired immune response finally occurs with Th1 polarity. The level of inflammation and adverse events depend on the TLR repertoire and the mode of response to their agonists in the relevant DC subsets, and could be predicted by assessing the levels of cytokines/chemokines and T cell proliferation in vaccinated subjects. The main differences in the mode of vaccine sought in infectious diseases and cancer are defined by whether it is prophylactic or therapeutic, whether it can deliver sufficient antigens to cDCs, and how it behaves in the microenvironment of the lesion. Adjuvant can be selected on a case-to-case basis.
• Assessing the pyrogenicity of whole influenza virus particle vaccine in cynomolgus macaques.
  Marumi Ohno; Masataka Sagata; Toshiki Sekiya; Naoki Nomura; Masashi Shingai; Masafumi Endo; Kazuhiko Kimachi; Saori Suzuki; Cong Thanh Nguyen; Misako Nakayama; Hirohito Ishigaki; Kazumasa Ogasawara; Yasushi Itoh; Yoichiro Kino; Hiroshi Kida
  Vaccine, 41, 3, 787, 794, 2023年01月16日, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, Among inactivated influenza vaccines, the whole virus particle vaccine (WPV) elicits superior priming responses to split virus vaccine (SV) in efficiently inducing humoral and cellular immunity. However, there is concern for undesired adverse events such as fever for WPV due to its potent immunogenicity. Therefore, this study investigated the febrile response induced by subcutaneous injection with quadrivalent inactivated influenza vaccines of good manufacturing grade for pharmaceutical or investigational products in cynomolgus macaques. Body temperature was increased by 1 °C-2 °C for 6-12 h after WPV administration at the first vaccination but not at the second shot, whereas SV did not affect body temperature at both points. Given the potent priming ability of WPV, WPV-induced fever may be attributed to immune responses that uniquely occur during priming. Since WPV-induced fever was blunted by pretreatment with indomethacin (a cyclooxygenase inhibitor), the febrile response by WPV is considered to depend on the increase in prostaglandins synthesized by cyclooxygenase. In addition, WPV, but not SV, induced the elevation of type I interferons and monocyte chemotactic protein 1 in the plasma; these factors may be responsible for pyrogenicity caused by WPV, as they can increase prostaglandins in the brain. Notably, sufficient antibody responses were acquired by half the amount of WPV without causing fever, suggesting that excessive immune responses to trigger the febrile response is not required for acquired immunity induction. Thus, we propose that WPV with a reduced antigen dose should be evaluated for potential clinical usage, especially in naïve populations.
• Field-deployable multiplex detection method of SARS-CoV-2 and influenza virus using loop-mediated isothermal amplification and DNA chromatography.
  Kyoko Hayashida; Alejandro Garcia; Lavel Chinyama Moonga; Tatsuki Sugi; Kodera Takuya; Mitsuo Kawase; Fumihiro Kodama; Atsushi Nagasaka; Nobuhisa Ishiguro; Ayato Takada; Masahiro Kajihara; Naganori Nao; Masashi Shingai; Hiroshi Kida; Yasuhiko Suzuki; William W Hall; Hirofumi Sawa; Junya Yamagishi
  PloS one, 18, 5, e0285861, 2023年, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, A novel multiplex loop-mediated isothermal amplification (LAMP) method combined with DNA chromatography was developed for the simultaneous detection of three important respiratory disease-causing viruses: severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), influenza A virus, and influenza B virus. Amplification was performed at a constant temperature, and a positive result was confirmed by a visible colored band. An in-house drying protocol with trehalose was used to prepare the dried format multiplex LAMP test. Using this dried multiplex LAMP test, the analytical sensitivity was determined to be 100 copies for each viral target and 100-1000 copies for the simultaneous detection of mixed targets. The multiplex LAMP system was validated using clinical COVID-19 specimens and compared with the real-time qRT-PCR method as a reference test. The determined sensitivity of the multiplex LAMP system for SARS-CoV-2 was 71% (95% CI: 0.62-0.79) for cycle threshold (Ct) ≤ 35 samples and 61% (95% CI: 0.53-0.69) for Ct ≤40 samples. The specificity was 99% (95%CI: 0.92-1.00) for Ct ≤35 samples and 100% (95%CI: 0.92-1.00) for the Ct ≤40 samples. The developed simple, rapid, low-cost, and laboratory-free multiplex LAMP system for the two major important respiratory viral diseases, COVID-19 and influenza, is a promising field-deployable diagnosis tool for the possible future 'twindemic, ' especially in resource-limited settings.
• Immunization with inactivated whole virus particle influenza virus vaccines improves the humoral response landscape in cynomolgus macaques
  Brendon Y. Chua; Toshiki Sekiya; Marios Koutsakos; Naoki Nomura; Louise C. Rowntree; Thi H. O. Nguyen; Hayley A. McQuilten; Marumi Ohno; Yuki Ohara; Tomohiro Nishimura; Masafumi Endo; Yasushi Itoh; Jennifer R. Habel; Kevin J. Selva; Adam K. Wheatley; Bruce D. Wines; P. Mark Hogarth; Stephen J. Kent; Amy W. Chung; David C. Jackson; Lorena E. Brown; Masashi Shingai; Katherine Kedzierska; Hiroshi Kida
  PLOS Pathogens, 18, 10, e1010891, e1010891, Public Library of Science (PLoS), 2022年10月07日, ［査読有り］
  研究論文（学術雑誌）, Although antibody-inducing split virus vaccines (SV) are currently the most effective way to combat seasonal influenza, their efficacy can be modest, especially in immunologically-naïve individuals. We investigated immune responses towards inactivated whole influenza virus particle vaccine (WPV) formulations, predicated to be more immunogenic, in a non-human primate model, as an important step towards clinical testing in humans. Comprehensive analyses were used to capture 46 immune parameters to profile how WPV-induced responses differed to those elicited by antigenically-similar SV formulations. Naïve cynomolgus macaques vaccinated with either monovalent or quadrivalent WPV consistently induced stronger antibody responses and hemagglutination inhibition (HI) antibody titres against vaccine-matched viruses compared to SV formulations, while acute reactogenic effects were similar. Responses in WPV-primed animals were further increased by boosting with the same formulation, conversely to modest responses after priming and boosting with SV. 28-parameter multiplex bead array defined key antibody features and showed that while both WPV and SV induced elevated IgG responses against A/H1N1 nucleoprotein, only WPV increased IgG responses against A/H1N1 hemagglutinin (HA) and HA-Stem, and higher IgA responses to A/H1N1-HA after each vaccine dose. Antibodies to A/H1N1-HA and HA-Stem that could engage FcγR2a and FcγR3a were also present at higher levels after one dose of WPV compared to SV and remained elevated after the second dose. Furthermore, WPV-enhanced antibody responses were associated with higher frequencies of HA-specific B-cells and IFN-γ-producing CD4⁺ T-cell responses. Our data additionally demonstrate stronger boosting of HI titres by WPV following prior infection and support WPV administered as a priming dose irrespective of the follow up vaccine for the second dose. Our findings thus show that compared to SV vaccination, WPV-induced humoral responses are significantly increased in scope and magnitude, advocating WPV vaccination regimens for priming immunologically-naïve individuals and also in the event of a pandemic outbreak.
• A high-affinity aptamer with base-appended base-modified DNA bound to isolated authentic SARS-CoV-2 strains wild-type and B.1.617.2 (delta variant).
  Hirotaka Minagawa; Hirofumi Sawa; Tomoko Fujita; Shintaro Kato; Asumi Inaguma; Miwako Hirose; Yasuko Orba; Michihito Sasaki; Koshiro Tabata; Naoki Nomura; Masashi Shingai; Yasuhiko Suzuki; Katsunori Horii
  Biochemical and biophysical research communications, 614, 207, 212, 2022年07月23日, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, Simple, highly sensitive detection technologies for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are crucial for the effective implementation of public health policies. We used the systematic evolution of ligands by exponential enrichment with a modified DNA library, including a base-appended base (uracil with a guanine base at its fifth position), to create an aptamer with a high affinity for the receptor-binding domain (RBD) of the SARS-CoV-2 spike glycoprotein. The aptamer had a dissociation constant of 1.2 and < 1 nM for the RBD and spike trimer, respectively. Furthermore, enzyme-linked aptamer assays confirmed that the aptamer binds to isolated authentic SARS-CoV-2 wild-type and B.1.617.2 (delta variant). The binding signal was larger that of commercially available anti-SARS-CoV-2 RBD antibody. Thus, this aptamer as a sensing element will enable the highly sensitive detection of SARS-CoV-2.
• Inactivated whole influenza virus particle vaccines induce neutralizing antibodies with an increase in immunoglobulin gene subclones of B-lymphocytes in cynomolgus macaques.
  Masanori Shiohara; Saori Suzuki; Shintaro Shichinohe; Hirohito Ishigaki; Misako Nakayama; Naoki Nomura; Masashi Shingai; Toshiki Sekiya; Marumi Ohno; Sayaka Iida; Naoko Kawai; Mamiko Kawahara; Junya Yamagishi; Kimihito Ito; Ryotarou Mitsumata; Tomio Ikeda; Kenji Motokawa; Tomoyoshi Sobue; Hiroshi Kida; Kazumasa Ogasawara; Yasushi Itoh
  Vaccine, 40, 30, 4026, 4037, 2022年06月26日, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, The All-Japan Influenza Vaccine Study Group has been developing a more effective vaccine than the current split vaccines for seasonal influenza virus infection. In the present study, the efficacy of formalin- and/or β-propiolactone-inactivated whole virus particle vaccines for seasonal influenza was compared to that of the current ether-treated split vaccines in a nonhuman primate model. The monovalent whole virus particle vaccines or split vaccines of influenza A virus (H1N1) and influenza B virus (Victoria lineage) were injected subcutaneously into naïve cynomolgus macaques twice. The whole virus particle vaccines induced higher titers of neutralizing antibodies against H1N1 influenza A virus and influenza B virus in the plasma of macaques than did the split vaccines. At challenge with H1N1 influenza A virus or influenza B virus, the virus titers in nasal swabs and the increases in body temperatures were lower in the macaques immunized with the whole virus particle vaccine than in those immunized with the split vaccine. Repertoire analyses of immunoglobulin heavy chain genes demonstrated that the number of B-lymphocyte subclones was increased in macaques after the 1st vaccination with the whole virus particle vaccine, but not with the split vaccine, indicating that the whole virus particle vaccine induced the activation of vaccine antigen-specific B-lymphocytes more vigorously than did the split vaccine at priming. Thus, the present findings suggest that the superior antibody induction ability of the whole virus particle vaccine as compared to the split vaccine is attributable to its stimulatory properties on the subclonal differentiation of antigen-specific B-lymphocytes.
• Inactivated Whole Virus Particle Influenza Vaccine Induces Anti-Neuraminidase Antibodies That May Contribute to Cross-Protection against Heterologous Virus Infection.
  Chimuka Handabile; Toshiki Sekiya; Naoki Nomura; Marumi Ohno; Tomomi Kawakita; Masashi Shingai; Hiroshi Kida
  Vaccines, 10, 5, 2022年05月19日, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, Despite the use of vaccines, seasonal influenza remains a risk to public health. We previously proposed the inactivated whole virus particle vaccine (WPV) as an alternative to the widely used split vaccine (SV) for the control of seasonal and pandemic influenza based on the superior priming potency of WPV to that of SV. In this study, we further examined and compared the immunological potency of monovalent WPV and SV of A/California/7/2009 (X-179A) (H1N1) pdm09 (CA/09) to generate immune responses against heterologous viruses, A/Singapore/GP1908/2015 (IVR-180) (H1N1) pdm09 (SG/15), and A/duck/Hokkaido/Vac-3/2007 (H5N1) (DH/07) in mice. Following challenge with a lethal dose of heterologous SG/15, lower virus titer in the lungs and milder weight loss were observed in WPV-vaccinated mice than in SV-vaccinated ones. To investigate the factors responsible for the differences in the protective effect against SG/15, the sera of vaccinated mice were analyzed by hemagglutination-inhibition (HI) and neuraminidase-inhibition (NI) assays to evaluate the antibodies induced against viral hemagglutinin (HA) and neuraminidase (NA), respectively. While the two vaccines induced similar levels of HI antibodies against SG/15 after the second vaccination, only WPV-vaccinated mice induced significantly higher titers of NI antibodies against the strain. Furthermore, given the significant elevation of NI antibody titers against DH/07, an H5N1 avian influenza virus, WPV was also demonstrated to induce NA-inhibiting antibodies that recognize NA of divergent strains. This could be explained by the higher conservation of epitopes of NA among strains than for HA. Taking these findings together, NA-specific antibodies induced by WPV may have contributed to better protection from infection with heterologous influenza virus SG/15, compared with SV. The present results indicate that WPV is an effective vaccine for inducing antibodies against both HA and NA of heterologous viruses and may be a useful vaccine to conquer vaccine strain mismatch.
• Leptospira Is an Environmental Bacterium That Grows in Waterlogged Soil.
  Yasutake Yanagihara; Sharon Y A M Villanueva; Naoki Nomura; Marumi Ohno; Toshiki Sekiya; Chimuka Handabile; Masashi Shingai; Hideaki Higashi; Shin-Ichi Yoshida; Toshiyuki Masuzawa; Nina G Gloriani; Mitsumasa Saito; Hiroshi Kida
  Microbiology spectrum, 10, 2, e0215721, 2022年04月27日, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, Leptospirosis is a zoonotic disease caused by infection with pathogenic leptospires. Consistent with recent studies by other groups, leptospires were isolated from 89 out of 110 (80.9%) soil or water samples from varied locations in the Philippines in our surveillance study, indicating that leptospires might have a life cycle that does not involve animal hosts. However, despite previous work, it has not been confirmed whether leptospires multiply in the soil environment under various experimental conditions. Given the fact that the case number of leptospirosis is increased after flood, we hypothesized that waterlogged soil, which mimics the postflooding environment, could be a suitable condition for growing leptospires. To verify this hypothesis, pathogenic and saprophytic leptospires were seeded in the bottles containing 2.5 times as much water as soil, and bacterial counts in the bottles were measured over time. Pathogenic and saprophytic leptospires were found to increase their number in waterlogged soil but not in water or soil alone. In addition, leptospires were reisolated from soil in closed tubes for as long as 379 days. These results indicate that leptospires are in a resting state in the soil and are able to proliferate with increased water content in the environment. This notion is strongly supported by observations that the case number of leptospirosis is significantly higher in rainy seasons and increased after flood. Therefore, we reached the following conclusion: environmental soil is a potential reservoir of leptospires. IMPORTANCE Since research on Leptospira has focused on pathogenic leptospires, which are supposed to multiply only in animal hosts, the life cycle of saprophytic leptospires has long been a mystery. This study demonstrates that both pathogenic and saprophytic leptospires multiply in the waterlogged soil, which mimics the postflooding environment. The present results potentially explain why leptospirosis frequently occurs after floods. Therefore, environmental soil is a potential reservoir of leptospires and leptospirosis is considered an environment-borne as well as a zoonotic disease. This is a significant report to reveal that leptospires multiply under environmental conditions, and this finding leads us to reconsider the ecology of leptospires.
• Abnormal blood coagulation and kidney damage in aged hamsters infected with severe acute respiratory syndrome coronavirus 2
  Marumi Ohno; Michihito Sasaki; Yasuko Orba; Toshiki Sekiya; Md Abdul Masum; Osamu Ichii; Tatsuya Sawamura; Akemi Kakino; Yasuhiko Suzuki; Hiroshi Kida; Hirofumi Sawa; Masashi Shingai
  Viruses, 13, 11, 2021年11月, ［査読有り］, ［最終著者, 責任著者］
  研究論文（学術雑誌）
• Defining the kinetic effects of infection with influenza virus A/PR8/34 (H1N1) on sphingosine-1-phosphate signaling in mice by targeted LC/MS.
  Divyavani Gowda; Marumi Ohno; Siddabasave Gowda B Gowda; Hitoshi Chiba; Masashi Shingai; Hiroshi Kida; Shu-Ping Hui
  Scientific reports, 11, 1, 20161, 20161, 2021年10月11日, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, Influenza remains a world-wide health concern, causing 290,000-600,000 deaths and up to 5 million cases of severe illnesses annually. Noticing the host factors that control biological responses, such as inflammatory cytokine secretion, to influenza virus infection is important for the development of novel drugs. Sphingosine-1-phosphate (S1P) is a bioactive sphingolipid metabolite and has essential biological functions in inflammation. However, the kinetic effects of influenza virus infection on physiological S1P levels and their signaling in multiple tissues remain unknown. In this study, we utilized a mouse model intranasally infected with 50 or 500 plaque forming units (PFU) of A/Puerto Rico/8/34 (H1N1; PR8) virus to investigate how S1P levels and expression of its regulating factors are affected by influenza virus infection by the liquid-chromatography/mass spectrometry and real-time PCR, respectively. The S1P level was significantly high in the plasma of mice infected with 500 PFU of the virus than that in control mice at 6 day-post-infection (dpi). Elevated gene expression of sphingosine kinase-1 (Sphk1), an S1P synthase, was observed in the liver, lung, white adipose tissue, heart, and aorta of infected mice. This could be responsible for the increased plasma S1P levels as well as the decrease in the hepatic S1P lyase (Sgpl1) gene in the infected mice. These results indicate modulation of S1P-signaling by influenza virus infection. Since S1P regulates inflammation and leukocyte migration, it must be worth trying to target this signaling to control influenza-associated symptoms.
• Critical role of oxidized LDL receptor-1 in intravascular thrombosis in a severe influenza mouse model.
  Marumi Ohno; Akemi Kakino; Toshiki Sekiya; Naoki Nomura; Masashi Shingai; Tatsuya Sawamura; Hiroshi Kida
  Scientific reports, 11, 1, 15675, 15675, 2021年08月03日, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, Although coagulation abnormalities, including microvascular thrombosis, are thought to contribute to tissue injury and single- or multiple-organ dysfunction in severe influenza, the detailed mechanisms have yet been clarified. This study evaluated influenza-associated abnormal blood coagulation utilizing a severe influenza mouse model. After infecting C57BL/6 male mice with intranasal applications of 500 plaque-forming units of influenza virus A/Puerto Rico/8/34 (H1N1; PR8), an elevated serum level of prothrombin fragment 1 + 2, an indicator for activated thrombin generation, was observed. Also, an increased gene expression of oxidized low-density lipoprotein (LDL) receptor-1 (Olr1), a key molecule in endothelial dysfunction in the progression of atherosclerosis, was detected in the aorta of infected mice. Body weight decrease, serum levels of cytokines and chemokines, viral load, and inflammation in the lungs of infected animals were similar between wild-type and Olr1 knockout (KO) mice. In contrast, the elevation of prothrombin fragment 1 + 2 levels in the sera and intravascular thrombosis in the lungs by PR8 virus infection were not induced in KO mice. Collectively, the results indicated that OLR1 is a critical host factor in intravascular thrombosis as a pathogeny of severe influenza. Thus, OLR1 is a promising novel therapeutic target for thrombosis during severe influenza.
• Potent priming by inactivated whole influenza virus particle vaccines is linked to viral RNA uptake into antigen presenting cells.
  Masashi Shingai; Naoki Nomura; Toshiki Sekiya; Marumi Ohno; Daisuke Fujikura; Chimuka Handabile; Ryosuke Omori; Yuki Ohara; Tomohiro Nishimura; Masafumi Endo; Kazuhiko Kimachi; Ryotarou Mitsumata; Tomio Ikeda; Hiroki Kitayama; Hironori Hatanaka; Tomoyoshi Sobue; Fumihito Muro; Saori Suzuki; Cong Thanh Nguyen; Hirohito Ishigaki; Misako Nakayama; Yuya Mori; Yasushi Itoh; Marios Koutsakos; Brendon Y Chua; Katherine Kedzierska; Lorena E Brown; David C Jackson; Kazumasa Ogasawara; Yoichiro Kino; Hiroshi Kida
  Vaccine, 39, 29, 3940, 3951, 2021年06月29日, ［査読有り］, ［筆頭著者］, ［国際誌］
  英語, 研究論文（学術雑誌）, Current detergent or ether-disrupted split vaccines (SVs) for influenza do not always induce adequate immune responses, especially in young children. This contrasts with the whole virus particle vaccines (WPVs) originally used against influenza that were immunogenic in both adults and children but were replaced by SV in the 1970s due to concerns with reactogenicity. In this study, we re-evaluated the immunogenicity of WPV and SV, prepared from the same batch of purified influenza virus, in cynomolgus macaques and confirmed that WPV is superior to SV in priming potency. In addition, we compared the ability of WPV and SV to induce innate immune responses, including the maturation of dendritic cells (DCs) in vitro. WPV stimulated greater production of inflammatory cytokines and type-I interferon in immune cells from mice and macaques compared to SV. Since these innate responses are likely triggered by the activation of pattern recognition receptors (PRRs) by viral RNA, the quantity and quality of viral RNA in each vaccine were assessed. Although the quantity of viral RNA was similar in the two vaccines, the amount of viral RNA of a length that can be recognized by PRRs was over 100-fold greater in WPV than in SV. More importantly, 1000-fold more viral RNA was delivered to DCs by WPV than by SV when exposed to preparations containing the same amount of HA protein. Furthermore, WPV induced up-regulation of the DC maturation marker CD86 on murine DCs, while SV did not. The present results suggest that the activation of antigen-presenting DCs, by PRR-recognizable viral RNA contained in WPV is responsible for the effective priming potency of WPV observed in naïve mice and macaques. WPV is thus recommended as an alternative option for seasonal influenza vaccines, especially for children.
• Selecting and Using the Appropriate Influenza Vaccine for Each Individual.
  Toshiki Sekiya; Marumi Ohno; Naoki Nomura; Chimuka Handabile; Masashi Shingai; David C Jackson; Lorena E Brown; Hiroshi Kida
  Viruses, 13, 6, 2021年05月24日, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, Despite seasonal influenza vaccines having been routinely used for many decades, influenza A virus continues to pose a global threat to humans, causing high morbidity and mortality each year. The effectiveness of the vaccine is largely dependent on how well matched the vaccine strains are with the circulating influenza virus strains. Furthermore, low vaccine efficacy in naïve populations such as young children, or in the elderly, who possess weakened immune systems, indicates that influenza vaccines need to be more personalized to provide broader community protection. Advances in both vaccine technologies and our understanding of influenza virus infection and immunity have led to the design of a variety of alternate vaccine strategies to extend population protection against influenza, some of which are now in use. In this review, we summarize the progress in the field of influenza vaccines, including the advantages and disadvantages of different strategies, and discuss future prospects. We also highlight some of the challenges to be faced in the ongoing effort to control influenza through vaccination.
• Updating the influenza virus library at Hokkaido University -It's potential for the use of pandemic vaccine strain candidates and diagnosis.
  Naoki Nomura; Keita Matsuno; Masashi Shingai; Marumi Ohno; Toshiki Sekiya; Ryosuke Omori; Yoshihiro Sakoda; Robert G Webster; Hiroshi Kida
  Virology, 557, 55, 61, 2021年05月, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, Genetic reassortment of influenza A viruses through cross-species transmission contributes to the generation of pandemic influenza viruses. To provide information on the ecology of influenza viruses, we have been conducting a global surveillance of zoonotic influenza and establishing an influenza virus library. Of 4580 influenza virus strains in the library, 3891 have been isolated from over 70 different bird species. The remaining 689 strains were isolated from humans, pigs, horses, seal, whale, and the environment. Phylogenetic analyses of the HA genes of the library isolates demonstrate that the library strains are distributed to all major known clusters of the H1, H2 and H3 subtypes of HA genes that are prevalent in humans. Since past pandemic influenza viruses are most likely genetic reassortants of zoonotic and seasonal influenza viruses, a vast collection of influenza A virus strains from various hosts should be useful for vaccine preparation and diagnosis for future pandemics.
• Immune profiling of influenza-specific B- and T-cell responses in macaques using flow cytometry-based assays.
  Marios Koutsakos; Toshiki Sekiya; Brendon Y Chua; Thi Hoang Oanh Nguyen; Adam K Wheatley; Jennifer A Juno; Marumi Ohno; Naoki Nomura; Yuki Ohara; Tomohiro Nishimura; Masafumi Endo; Saori Suzuki; Hirohito Ishigaki; Misako Nakayama; Cong T Nguyen; Yasushi Itoh; Masashi Shingai; Kazumasa Ogasawara; Yoichiro Kino; Stephen J Kent; David C Jackson; Lorena E Brown; Hiroshi Kida; Katherine Kedzierska
  Immunology and cell biology, 99, 1, 97, 106, 2021年01月, ［査読有り］, ［国際誌］
  英語, 研究論文（学術雑誌）, Influenza remains a significant global public health burden, despite substantial annual vaccination efforts against circulating virus strains. As a result, novel vaccine approaches are needed to generate long-lasting and universal broadly cross-reactive immunity against distinct influenza virus strains and subtypes. Several new vaccine candidates are currently under development and/or in clinical trials. The successful development of new vaccines requires testing in animal models, other than mice, which capture the complexity of the human immune system. Importantly, following vaccination or challenge, the assessment of adaptive immunity at the antigen-specific level is particularly informative. In this study, using peripheral blood mononuclear cells (PBMCs) from cynomolgus macaques, we describe detection methods and in-depth analyses of influenza virus-specific B cells by recombinant hemagglutinin probes and flow cytometry, as well as the detection of influenza virus-specific CD8+ and CD4+ T cells by stimulation with live influenza A virus and intracellular cytokine staining. We highlight the potential of these assays to be used with PBMCs from other macaque species, including rhesus macaques, pigtail macaques and African green monkeys. We also demonstrate the use of a human cytometric bead array kit in detecting inflammatory cytokines and chemokines from cynomolgus macaques to assess cytokine/chemokine milieu. Overall, the detection of influenza virus-specific B and T cells, together with inflammatory responses, as described in our study, provides useful insights for evaluating novel influenza vaccines. Our data deciphering immune responses toward influenza viruses can be also adapted to understanding immunity to other infections or vaccination approaches in macaque models.
• Influenza virus infection affects insulin signaling, fatty acid-metabolizing enzyme expressions, and the tricarboxylic acid cycle in mice
  Marumi Ohno; Toshiki Sekiya; Naoki Nomura; Taku ji Daito; Masashi Shingai; Hiroshi Kida
  Scientific Reports, 10, 1, Springer Science and Business Media LLC, 2020年12月, ［査読有り］
  研究論文（学術雑誌）
• Inactivated whole virus particle vaccine with potent immunogenicity and limited IL-6 production is ideal for influenza
  Toshiki Sekiya; Edin J Mifsud; Marumi Ohno; Naoki Nomura; Mayumi Sasada; Daisuke Fujikura; Takuji Daito; Masashi Shingai; Yuki Ohara; Tomohiro Nishimura; Masafumi Endo; Ryotarou Mitsumata; Tomio Ikeda; Hironori Hatanaka; Hiroki Kitayama; Kenji Motokawa; Tomoyoshi Sobue; Saori Suzuki; Yasushi Itoh; Lorena E Brown; Kazumasa Ogasawara; Yoichiro Kino; Hiroshi Kida
  Vaccine, 37, 15, 2158, 2166, Elsevier BV, 2019年03月, ［査読有り］
  英語, 研究論文（学術雑誌）
• Early antibody therapy can induce long-lasting immunity to SHIV
  Yoshiaki Nishimura; Rajeev Gautam; Tae-Wook Chun; Reza Sadjadpour; Kathryn E. Foulds; Masashi Shingai; Florian Klein; Anna Gazumyan; Jovana Golijanin; Mitzi Donaldson; Olivia K. Donau; Ronald J. Plishka; Alicia Buckler-White; Michael S. Seaman; Jeffrey D. Lifson; Richard A. . Koup; Anthony S. Fauci; Michel C. . Nussenzweig; Malcolm A. Martin
  NATURE, 543, 7646, 559, +, 2017年03月, ［査読有り］
  英語, 研究論文（学術雑誌）
• Development of mouse models for analysis of human virus infections
  Hiromi Takaki; Hiroyuki Oshiumi; Masashi Shingai; Misako Matsumoto; Tsukasa Seya
  MICROBIOLOGY AND IMMUNOLOGY, 61, 3-4, 107, 113, 2017年03月, ［査読有り］
  英語
• Antagonism of BST-2/Tetherin Is a Conserved Function of the Env Glycoprotein of Primary HIV-2 Isolates
  Chia-Yen Chen; Masashi Shingai; Sarah Welbourn; Malcolm A. Martin; Pedro Borrego; Nuno Taveira; Klaus Strebel
  JOURNAL OF VIROLOGY, 90, 24, 11062, 11074, 2016年12月, ［査読有り］
  英語, 研究論文（学術雑誌）
• The Expression of Functional Vpx during Pathogenic SIVmac Infections of Rhesus Macaques Suppresses SAMHD1 in CD4(+) Memory T Cells
  Masashi Shingai; Sarah Welbourn; Jason M. Brenchley; Priyamvada Acharya; Eri Miyagi; Ronald J. Plishka; Alicia Buckler-White; Peter D. Kwong; Yoshiaki Nishimura; Klaus Strebel; Malcolm A. Martin
  PLOS PATHOGENS, 11, 5, e1004928, 2015年05月, ［査読有り］, ［筆頭著者］
  英語, 研究論文（学術雑誌）
• Analysis of immunoglobulin transcripts and hypermutation following SHIVAD8 infection and protein-plus-adjuvant immunization
  Joseph R. Francica; Zizhang Sheng; Zhenhai Zhang; Yoshiaki Nishimura; Masashi Shingai; Akshaya Ramesh; Brandon F. Keele; Stephen D. Schmidt; Barbara J. Flynn; Sam Darko; Rebecca M. Lynch; Takuya Yamamoto; Rodrigo Matus-Nicodemos; David Wolinsky; Martha Nason; Nicholas M. Valiante; Padma Malyala; Ennio De Gregorio; Susan W. Barnett; Manmohan Singh; Derek T. O'Hagan; Richard A. Koup; John R. Mascola; Malcolm A. Martin; Thomas B. Kepler; Daniel C. Douek; Lawrence Shapiro; Robert A. Seder
  NATURE COMMUNICATIONS, 6, 6565, 2015年04月, ［査読有り］
  英語, 研究論文（学術雑誌）
• Enhanced HIV-1 immunotherapy by commonly arising antibodies that target virus escape variants
  Florian Klein; Lilian Nogueira; Yoshiaki Nishimura; Ganesh Phad; Anthony P. West; Ariel Halper-Stromberg; Joshua A. Horwitz; Anna Gazumyan; Cassie Liu; Thomas R. Eisenreich; Clara Lehmann; Gerd Faetkenheuer; Constance Williams; Masashi Shingai; Malcolm A. Martin; Pamela J. Bjorkman; Michael S. Seaman; Susan Zolla-Pazner; Gunilla B. Karlsson Hedestam; Michel C. Nussenzweig
  JOURNAL OF EXPERIMENTAL MEDICINE, 211, 12, 2361, 2372, 2014年11月, ［査読有り］
  英語, 研究論文（学術雑誌）
• Passive transfer of modest titers of potent and broadly neutralizing anti-HIV monoclonal antibodies block SHIV infection in macaques
  Masashi Shingai; Olivia K. Donau; Ronald J. Plishka; Alicia Buckler-White; John R. Mascola; Gary J. Nabel; Martha C. Nason; David Montefiori; Brian Moldt; Pascal Poignard; Ron Diskin; Pamela J. Bjorkman; Michael A. Eckhaus; Florian Klein; Hugo Mouquet; Julio Cesar Cetrulo Lorenzi; Anna Gazumyan; Dennis R. Burton; Michel C. Nussenzweig; Malcolm A. Martin; Yoshiaki Nishimura
  JOURNAL OF EXPERIMENTAL MEDICINE, 211, 10, 2061, 2074, 2014年09月, ［査読有り］, ［筆頭著者］
  英語, 研究論文（学術雑誌）
• MAVS-dependent IRF3/7 bypass of interferon β-induction restricts the response to measles infection in CD150Tg mouse bone marrow-derived dendritic cells.
  Takaki H; Honda K; Atarashi K; Kobayashi F; Ebihara T; Oshiumi H; Matsumoto M; Shingai M; Seya T
  Molecular immunology, 57, 2, 100, 109, 2014年02月, ［査読有り］
  英語, 研究論文（学術雑誌）
• The MyD88 Pathway in Plasmacytoid and CD4(+) Dendritic Cells Primarily Triggers Type I IFN Production against Measles Virus in a Mouse Infection Model
  Hiromi Takaki; Makoto Takeda; Maino Tahara; Masashi Shingai; Hiroyuki Oshiumi; Misako Matsumoto; Tsukasa Seya
  JOURNAL OF IMMUNOLOGY, 191, 9, 4740, 4747, 2013年11月, ［査読有り］
  英語, 研究論文（学術雑誌）
• Antibody-mediated immunotherapy of macaques chronically infected with SHIV suppresses viraemia.
  Shingai M; Nishimura Y; Klein F; Mouquet H; Donau OK; Plishka R; Buckler-White A; Seaman M; Piatak M Jr; Lifson JD; Dimitrov DS; Nussenzweig MC; Martin MA
  Nature, 503, 7475, 277, 280, 2013年11月, ［査読有り］, ［筆頭著者］
• Emergence of gp120 V3 Variants Confers Neutralization Resistance in an R5 Simian-Human Immunodeficiency Virus-Infected Macaque Elite Neutralizer That Targets the N332 Glycan of the Human Immunodeficiency Virus Type 1 Envelope Glycoprotein
  Reza Sadjadpour; Olivia K. Donau; Masashi Shingai; Alicia Buckler-White; Sandra Kao; Klaus Strebel; Yoshiaki Nishimura; Malcolm A. Martin
  JOURNAL OF VIROLOGY, 87, 15, 8798, 8804, 2013年08月, ［査読有り］
  英語, 研究論文（学術雑誌）
• Most rhesus macaques infected with the CCR5-tropic SHIVAD8 generate cross-reactive antibodies that neutralize multiple HIV-1 strains
  Masashi Shingai; Olivia K. Donau; Stephen D. Schmidt; Rajeev Gautam; Ronald J. Plishka; Alicia Buckler-White; Reza Sadjadpour; Wendy R. Lee; Celia C. LaBranche; David C. Montefiori; John R. Mascola; Yoshiaki Nishimura; Malcolm A. Martin
  PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 109, 48, 19769, 19774, 2012年11月, ［査読有り］, ［筆頭著者］
  英語, 研究論文（学術雑誌）
• Pathogenicity and Mucosal Transmissibility of the R5-Tropic Simian/Human Immunodeficiency Virus SHIVAD8 in Rhesus Macaques: Implications for Use in Vaccine Studies
  Rajeev Gautam; Yoshiaki Nishimura; Wendy R. Lee; Olivia Donau; Alicia Buckler-White; Masashi Shingai; Reza Sadjadpour; Stephen D. Schmidt; Celia C. LaBranche; Brandon F. Keele; David Montefiori; John R. Mascola; Malcolm A. Martin
  JOURNAL OF VIROLOGY, 86, 16, 8516, 8526, 2012年08月, ［査読有り］
  英語, 研究論文（学術雑誌）
• Rapid development of glycan-specific, broad, and potent anti-HIV-1 gp120 neutralizing antibodies in an R5 SIV/HIV chimeric virus infected macaque
  Laura M. Walker; Devin Sok; Yoshiaki Nishimura; Olivia Donau; Reza Sadjadpour; Rajeev Gautam; Masashi Shingai; Robert Pejchal; Alejandra Ramos; Melissa D. Simek; Yu Geng; Ian A. Wilson; Pascal Poignard; Malcolm A. Martin; Dennis R. Burton
  PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 108, 50, 20125, 20129, 2011年12月, ［査読有り］
  英語, 研究論文（学術雑誌）
• Some Human Immunodeficiency Virus Type 1 Vpu Proteins Are Able To Antagonize Macaque BST-2 In Vitro and In Vivo: Vpu-Negative Simian-Human Immunodeficiency Viruses Are Attenuated In Vivo
  Masashi Shingai; Takeshi Yoshida; Malcolm A. Martin; Klaus Strebel
  JOURNAL OF VIROLOGY, 85, 19, 9708, 9715, 2011年10月, ［査読有り］, ［筆頭著者］
  英語, 研究論文（学術雑誌）
• Recombination-Mediated Changes in Coreceptor Usage Confer an Augmented Pathogenic Phenotype in a Nonhuman Primate Model of HIV-1-Induced AIDS
  Yoshiaki Nishimura; Masashi Shingai; Wendy R. Lee; Reza Sadjadpour; Olivia K. Donau; Ronald Willey; Jason M. Brenchley; Ranjini Iyengar; Alicia Buckler-White; Tatsuhiko Igarashi; Malcolm A. Martin
  JOURNAL OF VIROLOGY, 85, 20, 10617, 10626, 2011年10月, ［査読有り］
  英語, 研究論文（学術雑誌）
• Strain-to-strain difference of V protein of measles virus affects MDA5-mediated IFN-β-inducing potential.
  Takaki H; Watanabe Y; Shingai M; Oshiumi H; Matsumoto M; Seya T
  Molecular immunology, 48, 4, 497, 504, 2011年01月, ［査読有り］
  英語, 研究論文（学術雑誌）
• Generation of the Pathogenic R5-Tropic Simian/Human Immunodeficiency Virus SHIVAD8 by Serial Passaging in Rhesus Macaques
  Yoshiaki Nishimura; Masashi Shingai; Ronald Willey; Reza Sadjadpour; Wendy R. Lee; Charles R. Brown; Jason M. Brenchley; Alicia Buckler-White; Rahel Petros; Michael Eckhaus; Victoria Hoffman; Tatsuhiko Igarashi; Malcolm A. Martin
  JOURNAL OF VIROLOGY, 84, 9, 4769, 4781, 2010年05月, ［査読有り］
  英語, 研究論文（学術雑誌）
• Initiation of Antiretroviral Therapy 48 Hours after Infection with Simian Immunodeficiency Virus Potently Suppresses Acute-Phase Viremia and Blocks the Massive Loss of Memory CD4(+) T Cells but Fails To Prevent Disease
  Makoto Kubo; Yoshiaki Nishimura; Masashi Shingai; Wendy Lee; Jason Brenchley; Bernard Lafont; Alicia Buckler-White; Tatsuhiko Igarashi; Malcolm A. Martin
  JOURNAL OF VIROLOGY, 83, 14, 7099, 7108, 2009年07月, ［査読有り］
  英語, 研究論文（学術雑誌）
• Soluble G protein of respiratory syncytial virus inhibits Toll-like receptor 3/4-mediated IFN-beta induction
  Masashi Shingai; Masahiro Azuma; Takashi Ebihara; Miwa Sasai; Kenji Funami; Minoru Ayata; Hisashi Ogura; Hiroyuki Tsutsumi; Misako Matsumoto; Tsukasa Seya
  INTERNATIONAL IMMUNOLOGY, 20, 9, 1169, 1180, 2008年09月, ［査読有り］, ［筆頭著者］
  英語, 研究論文（学術雑誌）
• Hepatitis C virus-infected hepatocytes extrinsically modulate dendritic cell maturation to activate T cells and natural killer cells
  Takashi Ebihara; Masashi Shingai; Misako Matsumoto; Takaji Wakita; Tsukasa Seya
  HEPATOLOGY, 48, 1, 48, 58, 2008年07月, ［査読有り］
  英語, 研究論文（学術雑誌）
• Combinational recognition of bacterial lipoproteins and peptidoglycan by chicken Toll-like receptor 2 subfamily
  Megumi Higuchi; Aya Matsuo; Masashi Shingai; Kyoko Shida; Akihiro Ishii; Kenji Funami; Yasuhiko Suzuki; Hiroyuki Oshiumi; Misako Matsumoto; Tsukasa Seya
  DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY, 32, 2, 147, 155, 2008年, ［査読有り］
  英語, 研究論文（学術雑誌）
• Effect of the alterations in the fusion protein of measles virus isolated from brains of patients with subacute sclerosing panencephalitis on syncytium formation
  Minoru Ayata; Masashi Shingai; Xiaojun Ning; Misako Matsumoto; Tsukasa Seya; Sanae Otani; Toshiyuki Seto; Shinji Ohgimoto; Hisashi Ogura
  VIRUS RESEARCH, 130, 1-2, 260, 268, 2007年12月, ［査読有り］
  英語, 研究論文（学術雑誌）
• Differential type IIFN-Inducing abilities of wild-type versus vaccine strains of measles virus
  Masashi Shingai; Takashi Ebihara; Nasim A. Begum; Atsushi Kato; Toshiki Honma; Kenji Matsumoto; Hirohisa Saito; Hisashi Ogura; Misako Matsumoto; Tsukasa Seya
  JOURNAL OF IMMUNOLOGY, 179, 9, 6123, 6133, 2007年11月, ［査読有り］, ［筆頭著者］
  英語, 研究論文（学術雑誌）
• Induction of NKG2D ligands on human dendritic cells by TLR ligand stimulation and RNA virus infection
  Takashi Ebihara; Hisayo Masuda; Takashi Akazawa; Masashi Shingai; Hideaki Kikuta; Tadashi Ariga; Misako Matsumoto; Tsukasa Seya
  INTERNATIONAL IMMUNOLOGY, 19, 10, 1145, 1155, 2007年10月, ［査読有り］
  英語, 研究論文（学術雑誌）
• Tumor immunotherapy using bone marrow-derived dendritic cells overexpressing Toll-like receptor adaptors
  Takashi Akazawa; Masashi Shingai; Mwa Sasai; Takashi Ebihara; Norimitsu Inoue; Misako Matsumoto; Tsukasa Seya
  FEBS LETTERS, 581, 18, 3334, 3340, 2007年07月, ［査読有り］
  英語, 研究論文（学術雑誌）
• Antitumor NK activation induced by the Toll-like receptor 3-TICAM-1 (TRIF) pathway in myeloid dendritic cells
  Takashi Akazawa; Takashi Ebihara; Manabu Okuno; Yu Okuda; Masashi Shingai; Kunio Tsujimura; Toshitada Takahashi; Masahito Ikawa; Masaru Okabe; Norimitsu Inoue; Miki Okamoto-Tanaka; Hiroyoshi Ishizaki; Jun Miyoshi; Misako Matsumoto; Tsukasa Seya
  PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 104, 1, 252, 257, 2007年01月, ［査読有り］
  英語, 研究論文（学術雑誌）
• NAK-associated protein 1 participates in both the TLR3 and the cytoplasmic pathways in type IIFN induction
  Miwa Sasai; Masashi Shingai; Kenji Funami; Mitsutoshi Yoneyama; Takashi Fujita; Misako Matsumoto; Tsukasa Seya
  JOURNAL OF IMMUNOLOGY, 177, 12, 8676, 8683, 2006年12月, ［査読有り］
  英語, 研究論文（学術雑誌）
• ウイルス感染と宿主応答・宿主側の要因 感染樹状細胞における麻疹ウイルスによるI型IFN誘導メカニズム(A mechanism by which measles virus induces Type-I IFN in infected dendritic cells)
  Shingai Masashi; Ebihara Takashi; Matsumoto Misako; Seya Tsukasa
  日本免疫学会総会・学術集会記録, 36, 207, 207, (NPO)日本免疫学会, 2006年11月
  英語
• NK細胞とMHC受容体 TLRリガンド刺激及びRNAウイルス感染によるヒト樹状細胞上のNKG2Dリガンドの誘導(NKG2D ligands are induced on human dendritic cells by TLR ligand stimulation and RNA virus infection)
  Ebihara Takashi; Shingai Masashi; Akazawa Takashi; Matsumoto Misako; Seya Tsukasa
  日本免疫学会総会・学術集会記録, 36, 253, 253, (NPO)日本免疫学会, 2006年11月
  英語
• Wild-type measles virus infection in human CD46/CD150-transgenic mice: CD11c-positive dendritic cells establish systemic viral infection
  M Shingai; N Inoue; T Okuno; M Okabe; T Akazawa; Y Miyamoto; M Ayata; K Honda; M Kurita-Taniguchi; M Matsumoto; H Ogura; T Taniguchi; T Seya
  JOURNAL OF IMMUNOLOGY, 175, 5, 3252, 3261, 2005年09月, ［査読有り］, ［筆頭著者］
  英語, 研究論文（学術雑誌）
• [Immunologic tests: Membrane cofactor protein (MCP, CD46)].
  Shingai M; Seya T
  Nihon rinsho. Japanese journal of clinical medicine, 63 Suppl 7, 95, 97, 2005年07月, ［査読有り］
• Dendritic cell maturation induced by muramyl dipeptide (MDP) derivatives: Monoacylated MDP confers TLR2/TLR4 activation
  J Uehori; K Fukase; T Akazawa; S Uematsu; S Akira; K Funami; M Shingai; M Matsumoto; Azuma, I; K Toyoshima; S Kusumoto; T Seya
  JOURNAL OF IMMUNOLOGY, 174, 11, 7096, 7103, 2005年06月, ［査読有り］
  英語, 研究論文（学術雑誌）
• [Toll-like receptors that sense viral infection].
  Seya T; Shingai M; Matsumoto M
  Uirusu, 54, 1, 1, 8, 日本ウイルス学会, 2004年06月, ［査読有り］
  日本語, TLRファミリーはロイシンリッチリピート (LRR) の細胞外領域と細胞内のTIR (Toll/IL-1 R homology) ドメインからなる. TLRはリガンドを認識すると細胞内のTIRドメインへアダプター分子MyD88などをリクルートし, NF-κBとMAP kinaseを活性化し, 最終的にサイトカインやケモカインなどを産生誘導する. 一方, 骨髄性樹状細胞 (mDC) ではTLRによるIFN-β産生やIFN誘導遺伝子の発現, 樹状細胞の成熟化などが誘起されるが, これらはアダプター分子, MyD88非依存性に起こる. TLRのIFN-β産生系としてはdsRNA刺激によるTLR3依存性経路, LPS刺激によるTLR4を介したIFN-β産生経路などがある. リンパ球様樹状細胞 (pDC) ではTLR7/TLR9刺激でIFN-αがMyD88依存性に誘導される. 我々は, 本稿でTLR3, TLR4を介したIFN-β産生に関与するアダプター分子の構造と機能を総説し, 新規のIFN-β誘導経路に言及する. 併せてウイルス成分認識性のTLRがtype I IFNを誘導するシグナル経路, これらの経路とウイルス感染が誘起する抗ウイルスの細胞応答の連携を概説する.
• Infection of different cell lines of neural origin with subacute sclerosing panencephalitis (SSPE) virus
  H Ishida; M Ayata; M Shingai; Matsunaga, I; Y Seto; Y Katayama; N Iritani; T Seya; Y Yanagi; O Matsuoka; T Yamano; H Ogura
  MICROBIOLOGY AND IMMUNOLOGY, 48, 4, 277, 287, 2004年, ［査読有り］
  英語, 研究論文（学術雑誌）
• Subcellular localization of toll-like receptor 3 in human dendritic cells
  M Matsumoto; K Funami; M Tanabe; H Oshiumi; M Shingai; Y Seto; A Yamamoto; T Seya
  JOURNAL OF IMMUNOLOGY, 171, 6, 3154, 3162, 2003年09月, ［査読有り］
  英語, 研究論文（学術雑誌）
• Receptor use by vesicular stomatitis virus pseudotypes with glycoproteins of defective variants of measles virus isolated from brains of patients with subacute sclerosing panencephalitis
  M Shingai; M Ayata; H Ishida; Matsunaga, I; Y Katayama; T Seya; H Tatsuo; Y Yanagi; H Ogura
  JOURNAL OF GENERAL VIROLOGY, 84, Pt 8, 2133, 2143, 2003年08月, ［査読有り］, ［筆頭著者］
  英語, 研究論文（学術雑誌）
• Mutations affecting transcriptional termination in the P gene end of subacute sclerosing panencephalitis viruses
  M Ayata; K Komase; M Shingai; Matsunaga, I; Y Katayama; H Ogura
  JOURNAL OF VIROLOGY, 76, 24, 13062, 13068, 2002年12月, ［査読有り］
  英語, 研究論文（学術雑誌）
• Alterations and diversity in the cytoplasmic tail of the fusion protein of subacute sclerosing panencephalitis virus strains isolated in Osaka, Japan
  Ning, X; M Ayata; M Kimura; K Komase; K Furukawa; T Seto; N Ito; M Shingai; Matsunaga, I; T Yamano; H Ogura
  VIRUS RESEARCH, 86, 1-2, 123, 131, 2002年06月, ［査読有り］
  英語, 研究論文（学術雑誌）
• Comparison of the neuropathogenicity of two SSPE sibling viruses of the Osaka-2 strain isolated with Vero and B95a cells
  N Ito; M Ayata; M Shingai; K Furukawa; T Seto; Matsunaga, I; M Muraoka; H Ogura
  JOURNAL OF NEUROVIROLOGY, 8, 1, 6, 13, 2002年02月, ［査読有り］
  英語, 研究論文（学術雑誌）
• Cell surface expression of immature H glycoprotein in measles virus-infected cells
  Ogura H; Matsunaga I; Takano Y; Ning XJ; Ayata M; Tanaka K; Seto T; Furukawa K; Ito N; Shingai M; Kimura T; Ichihara K; Kubo H; Murakami T
  VIRUS RESEARCH, 66, 2, 187, 196, 2000年02月, ［査読有り］
  英語

• インフルエンザウイルスA/Singapore/GP1908/2015(H1N1)ヘマグルチニンの単粒子構造解析
  小林颯太; 関屋俊輝; 喜多俊介; 新開大史; 喜田宏; 前仲勝実; 前仲勝実; 前仲勝実; 前仲勝実, 日本生体防御学会学術総会講演抄録集, 35th (Web), 2024年
• インフルエンザウイルスライブラリーを活用したパンデミックインフルエンザ対策
  野村直樹; 新開大史; 松野啓太; 関屋俊輝; 関屋俊輝; 大野円実; 磯田典和; 迫田義博; 喜田宏; 喜田宏, 日本獣医学会学術集会講演要旨集, 164th (CD-ROM), 2021年
• 不活化インフルエンザウイルス全粒子ワクチンの非臨床試験
  新開大史; 新開大史; 野村直樹; 関屋俊輝; 関屋俊輝; 大野円実; 笹田万友美; 大東卓史; 鈴木紗織; 石垣宏仁; 伊藤靖; 小笠原一誠; 小笠原一誠; 喜田宏; 喜田宏; 喜田宏, 日本ワクチン学会学術集会プログラム・抄録集, 22nd, 2018年
• サルモデルを用いた季節性インフルエンザに対する不活化全粒子ワクチンの有効性評価
  伊藤靖; 鈴木紗織; 石垣宏仁; 仲山美沙子; 新開大史; 新開大史; 大東卓史; 野村直樹; 喜田宏; 喜田宏; 喜田宏; 小笠原一誠; 小笠原一誠, 日本ワクチン学会学術集会プログラム・抄録集, 22nd, 2018年
• 異種間ウイルス伝播における自然免疫の役割 : 麻疹ウイルス感染の成立における Type-I IFN の重要性
  新開 大史; 松本 美佐子; 瀬谷 司, 臨床とウイルス, 35, 1, 21, 29, 2007年03月31日
  日本語
• NAP1 but not TICAM-1 participates in respiratory syncytial virus (RSV)-mediated interferon induction
  SHINGAI M., Int. J. Mol. Med., 2005年

• 感染症学特別研究Ⅰ, 2024年, 博士後期課程, 国際感染症学院
• 大学院共通授業科目（一般科目）：自然科学・応用科学, 2024年, 修士課程, 大学院共通科目
• 感染症学特別演習, 2024年, 博士後期課程, 国際感染症学院
• 人獣共通感染症制御学特論, 2024年, 博士後期課程, 国際感染症学院
• 感染症学特別研究ⅡA, 2024年, 博士後期課程, 国際感染症学院
• 生体防御学特論, 2024年, 博士後期課程, 国際感染症学院
• 感染症学特別研究ⅡB, 2024年, 博士後期課程, 国際感染症学院

• インフルエンザウイルス感染に伴う抗原原罪現象の誘導機序解明
  科学研究費助成事業
  2018年04月 - 2021年03月
  新開 大史; 喜田 宏
  インフルエンザウイルスの感染モデルマウスを用いて、抗原原罪が実際に起こることを確認した。免疫グロブリンのレパトア解析を行ったところ、PR8感染群は非感染群に比べてレパトアの多様性が高いことがわかった。これは、感染によってより多様な免疫グロブリンが誘導されたことを示唆している。また、免疫グロブリンのCDRH3領域を解析した結果、インフルエンザウイルスに特異的なCDRH3のアミノ酸配列を発見した。現在、免疫グロブリンレパトア解析により、コントロールマウスと抗原原罪マウスで誘導される抗体のCDRH3配列パターンの違いを解析している。
  日本学術振興会, 基盤研究(C), 北海道大学, 18K07135
• 麻疹ウイルス感染モデルを用いた樹状細胞からの麻疹ウイルス免疫抑制機構の解明
  科学研究費助成事業
  2006年 - 2007年
  新開 大史
  本研究は麻疹ウイルス感染モデルマウスを用い、麻疹ウイルス感染時に起こる免疫抑制のメカニズムの解明を目指す。
  リンパ球側の要因:各種Tgマウス(hCD46/hCD150 tg,hCD46 tg,hCD150 tg,non-tg,hCD46/hCD150 tg IFNARko,hCD46 tg IFNARko,hCD150 tg IFNARko,non tg IFNARko)に麻疹ウイルス(MV323)株を4×105 PFU静脈内投与し、4日後、マウスの脾臓よりリンパ球を回収、PHAにて刺激した。3H-チミジンの取り込みにてリンパ球の増殖を測定したところ、IFNARkoマウスにて有意にマイトージェン刺激(PHA)に対する応答性が低下した。同様にCD4陽性、CD8陽性細胞それぞれもIFNAR依存性に増殖抑制が確認できた。野外ウイルス株MV323GFP(GFP発現麻疹ウイルス)の蛍光探索から、免疫抑制が起こっているTリンパ球には麻疹ウイルスが感染していないことを明らかにした。以上より、現在麻疹ウイルス感染モデルマウスにおいて、T細胞の免疫抑制がヒトと同様に起こるが、T細胞にはウイルスが直接感染していないことが明らかとなった。
  樹上細胞側の要因:各種Tgマウスから、骨髄性樹状細胞を既報によって調整し、抗原提示機能、サイトカイン誘導能、CTL活性化能、NK活性可能などを個別に検定した。IFN誘導が感染応答に重要であったが、これらのマウスではIFNが殆ど誘導されなかった。そこで、CD150陽性でIRF-3,IRF-7の欠損マウスで免疫応答を調べた。結果はIRF-3がIFN誘導のみならず抗ウイルスNK活性化の誘導にも必須の役割を果たしていることが判明した。IRF3欠損マウスでIFNAR欠損マウスと同様の易感染性が見られることも判明した。このマウスにおける抗原提示細胞の機能を検討する予定である。
  日本学術振興会, 若手研究(B), 北海道大学, 18790317