2022年09月, 日本獣医学会, 獣医学奨励賞　　　　　　　　　　　　　　　
鳥インフルエンザ及び牛伝染性リンパ腫ウイルス感染症の病態解析

Virological characteristics of a SARS-CoV-2-related bat coronavirus, BANAL-20-236.
Shigeru Fujita, Arnon Plianchaisuk, Sayaka Deguchi, Hayato Ito, Naganori Nao, Lei Wang, Hesham Nasser, Tomokazu Tamura, Izumi Kimura, Yukie Kashima, Rigel Suzuki, Saori Suzuki, Izumi Kida, Masumi Tsuda, Yoshitaka Oda, Rina Hashimoto, Yukio Watanabe, Keiya Uriu, Daichi Yamasoba, Ziyi Guo, Alfredo A Hinay Jr, Yusuke Kosugi, Luo Chen, Lin Pan, Yu Kaku, Hin Chu, Flora Donati, Sarah Temmam, Marc Eloit, Yuki Yamamoto, Tetsuharu Nagamoto, Hiroyuki Asakura, Mami Nagashima, Kenji Sadamasu, Kazuhisa Yoshimura, Yutaka Suzuki, Jumpei Ito, Terumasa Ikeda, Shinya Tanaka, Keita Matsuno, Takasuke Fukuhara, Kazuo Takayama, Kei Sato
EBioMedicine, 104, 105181, 105181, 2024年06月, ［国際誌］
英語, 研究論文（学術雑誌）, BACKGROUND: Although several SARS-CoV-2-related coronaviruses (SC2r-CoVs) were discovered in bats and pangolins, the differences in virological characteristics between SARS-CoV-2 and SC2r-CoVs remain poorly understood. Recently, BANAL-20-236 (B236) was isolated from a rectal swab of Malayan horseshoe bat and was found to lack a furin cleavage site (FCS) in the spike (S) protein. The comparison of its virological characteristics with FCS-deleted SARS-CoV-2 (SC2ΔFCS) has not been conducted yet. METHODS: We prepared human induced pluripotent stem cell (iPSC)-derived airway and lung epithelial cells and colon organoids as human organ-relevant models. B236, SARS-CoV-2, and artificially generated SC2ΔFCS were used for viral experiments. To investigate the pathogenicity of B236 in vivo, we conducted intranasal infection experiments in hamsters. FINDINGS: In human iPSC-derived airway epithelial cells, the growth of B236 was significantly lower than that of the SC2ΔFCS. A fusion assay showed that the B236 and SC2ΔFCS S proteins were less fusogenic than the SARS-CoV-2 S protein. The infection experiment in hamsters showed that B236 was less pathogenic than SARS-CoV-2 and even SC2ΔFCS. Interestingly, in human colon organoids, the growth of B236 was significantly greater than that of SARS-CoV-2. INTERPRETATION: Compared to SARS-CoV-2, we demonstrated that B236 exhibited a tropism toward intestinal cells rather than respiratory cells. Our results are consistent with a previous report showing that B236 is enterotropic in macaques. Altogether, our report strengthens the assumption that SC2r-CoVs in horseshoe bats replicate primarily in the intestinal tissues rather than respiratory tissues. FUNDING: This study was supported in part by AMED ASPIRE (JP23jf0126002, to Keita Matsuno, Kazuo Takayama, and Kei Sato); AMED SCARDA Japan Initiative for World-leading Vaccine Research and Development Centers "UTOPIA" (JP223fa627001, to Kei Sato), AMED SCARDA Program on R&D of new generation vaccine including new modality application (JP223fa727002, to Kei Sato); AMED SCARDA Hokkaido University Institute for Vaccine Research and Development (HU-IVReD) (JP223fa627005h0001, to Takasuke Fukuhara, and Keita Matsuno); AMED Research Program on Emerging and Re-emerging Infectious Diseases (JP21fk0108574, to Hesham Nasser; JP21fk0108493, to Takasuke Fukuhara; JP22fk0108617 to Takasuke Fukuhara; JP22fk0108146, to Kei Sato; JP21fk0108494 to G2P-Japan Consortium, Keita Matsuno, Shinya Tanaka, Terumasa Ikeda, Takasuke Fukuhara, and Kei Sato; JP21fk0108425, to Kazuo Takayama and Kei Sato; JP21fk0108432, to Kazuo Takayama, Takasuke Fukuhara and Kei Sato; JP22fk0108534, Terumasa Ikeda, and Kei Sato; JP22fk0108511, to Yuki Yamamoto, Terumasa Ikeda, Keita Matsuno, Shinya Tanaka, Kazuo Takayama, Takasuke Fukuhara, and Kei Sato; JP22fk0108506, to Kazuo Takayama and Kei Sato); AMED Research Program on HIV/AIDS (JP22fk0410055, to Terumasa Ikeda; and JP22fk0410039, to Kei Sato); AMED Japan Program for Infectious Diseases Research and Infrastructure (JP22wm0125008 to Keita Matsuno); AMED CREST (JP21gm1610005, to Kazuo Takayama; JP22gm1610008, to Takasuke Fukuhara; JST PRESTO (JPMJPR22R1, to Jumpei Ito); JST CREST (JPMJCR20H4, to Kei Sato); JSPS KAKENHI Fund for the Promotion of Joint International Research (International Leading Research) (JP23K20041, to G2P-Japan Consortium, Keita Matsuno, Takasuke Fukuhara and Kei Sato); JST SPRING (JPMJSP2108 to Shigeru Fujita); JSPS KAKENHI Grant-in-Aid for Scientific Research C (22K07103, to Terumasa Ikeda); JSPS KAKENHI Grant-in-Aid for Scientific Research B (21H02736, to Takasuke Fukuhara); JSPS KAKENHI Grant-in-Aid for Early-Career Scientists (22K16375, to Hesham Nasser; 20K15767, to Jumpei Ito); JSPS Core-to-Core Program (A. Advanced Research Networks) (JPJSCCA20190008, to Kei Sato); JSPS Research Fellow DC2 (22J11578, to Keiya Uriu); JSPS Research Fellow DC1 (23KJ0710, to Yusuke Kosugi); JSPS Leading Initiative for Excellent Young Researchers (LEADER) (to Terumasa Ikeda); World-leading Innovative and Smart Education (WISE) Program 1801 from the Ministry of Education, Culture, Sports, Science and Technology (MEXT) (to Naganori Nao); Ministry of Health, Labour and Welfare (MHLW) under grant 23HA2010 (to Naganori Nao and Keita Matsuno); The Cooperative Research Program (Joint Usage/Research Center program) of Institute for Life and Medical Sciences, Kyoto University (to Kei Sato); International Joint Research Project of the Institute of Medical Science, the University of Tokyo (to Terumasa Ikeda and Takasuke Fukuhara); The Tokyo Biochemical Research Foundation (to Kei Sato); Takeda Science Foundation (to Terumasa Ikeda and Takasuke Fukuhara); Mochida Memorial Foundation for Medical and Pharmaceutical Research (to Terumasa Ikeda); The Naito Foundation (to Terumasa Ikeda); Hokuto Foundation for Bioscience (to Tomokazu Tamura); Hirose Foundation (to Tomokazu Tamura); and Mitsubishi Foundation (to Kei Sato).

Comparative pathogenicity of SARS-CoV-2 Omicron subvariants including BA.1, BA.2, and BA.5
Tomokazu Tamura, Daichi Yamasoba, Yoshitaka Oda, Jumpei Ito, Tomoko Kamasaki, Naganori Nao, Rina Hashimoto, Yoichiro Fujioka, Rigel Suzuki, Lei Wang, Hayato Ito, Yukie Kashima, Izumi Kimura, Mai Kishimoto, Masumi Tsuda, Hirofumi Sawa, Kumiko Yoshimatsu, Yuki Yamamoto, Tetsuharu Nagamoto, Jun Kanamune, Yutaka Suzuki, Yusuke Ohba, Saori Suzuki, Marie Kato, Zannatul Ferdous, Hiromi Mouri, Kenji Shishido, Naoko Misawa, Keiya Uriu, Yusuke Kosugi, Shigeru Fujita, Mai Suganami, Mika Chiba, Ryo Yoshimura, So Nakagawa, Jiaqi Wu, Akifumi Takaori-Kondo, Kotaro Shirakawa, Kayoko Nagata, Yasuhiro Kazuma, Ryosuke Nomura, Yoshihito Horisawa, Yusuke Tashiro, Yugo Kawai, Takao Hashiguchi, Tateki Suzuki, Kanako Kimura, Jiei Sasaki, Yukari Nakajima, Ayaka Sakamoto, Naoko Yasuhara, Takashi Irie, Ryoko Kawabata, Terumasa Ikeda, Hesham Nasser, Ryo Shimizu, Monira Begum, Otowa Takahashi, Kimiko Ichihara, Takamasa Ueno, Chihiro Motozono, Mako Toyoda, Akatsuki Saito, Yuri L. Tanaka, Erika P. Butlertanaka, Maya Shofa, Kaori Tabata, Isao Yokota, Keita Matsuno, Kazuo Takayama, Shinya Tanaka, Kei Sato, Takasuke Fukuhara
Communications Biology, 6, 1, Springer Science and Business Media LLC, 2023年07月24日
研究論文（学術雑誌）, Abstract

The unremitting emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants necessitates ongoing control measures. Given its rapid spread, the new Omicron subvariant BA.5 requires urgent characterization. Here, we comprehensively analyzed BA.5 with the other Omicron variants BA.1, BA.2, and ancestral B.1.1. Although in vitro growth kinetics of BA.5 was comparable among the Omicron subvariants, BA.5 was much more fusogenic than BA.1 and BA.2. Airway-on-a-chip analysis showed that, among Omicron subvariants, BA.5 had enhanced ability to disrupt the respiratory epithelial and endothelial barriers. Furthermore, in our hamster model, in vivo pathogenicity of BA.5 was slightly higher than that of the other Omicron variants and less than that of ancestral B.1.1. Notably, BA.5 gains efficient virus spread compared with BA.1 and BA.2, leading to prompt immune responses. Our findings suggest that BA.5 has low pathogenicity compared with the ancestral strain but enhanced virus spread /inflammation compared with earlier Omicron subvariants.

Virological characteristics of the SARS-CoV-2 XBB variant derived from recombination of two Omicron subvariants.
Tomokazu Tamura, Jumpei Ito, Keiya Uriu, Jiri Zahradnik, Izumi Kida, Yuki Anraku, Hesham Nasser, Maya Shofa, Yoshitaka Oda, Spyros Lytras, Naganori Nao, Yukari Itakura, Sayaka Deguchi, Rigel Suzuki, Lei Wang, Mst Monira Begum, Shunsuke Kita, Hisano Yajima, Jiei Sasaki, Kaori Sasaki-Tabata, Ryo Shimizu, Masumi Tsuda, Yusuke Kosugi, Shigeru Fujita, Lin Pan, Daniel Sauter, Kumiko Yoshimatsu, Saori Suzuki, Hiroyuki Asakura, Mami Nagashima, Kenji Sadamasu, Kazuhisa Yoshimura, Yuki Yamamoto, Tetsuharu Nagamoto, Gideon Schreiber, Katsumi Maenaka, Takao Hashiguchi, Terumasa Ikeda, Takasuke Fukuhara, Akatsuki Saito, Shinya Tanaka, Keita Matsuno, Kazuo Takayama, Kei Sato
Nature communications, 14, 1, 2800, 2800, 2023年05月16日, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, In late 2022, SARS-CoV-2 Omicron subvariants have become highly diversified, and XBB is spreading rapidly around the world. Our phylogenetic analyses suggested that XBB emerged through the recombination of two cocirculating BA.2 lineages, BJ.1 and BM.1.1.1 (a progeny of BA.2.75), during the summer of 2022. XBB.1 is the variant most profoundly resistant to BA.2/5 breakthrough infection sera to date and is more fusogenic than BA.2.75. The recombination breakpoint is located in the receptor-binding domain of spike, and each region of the recombinant spike confers immune evasion and increases fusogenicity. We further provide the structural basis for the interaction between XBB.1 spike and human ACE2. Finally, the intrinsic pathogenicity of XBB.1 in male hamsters is comparable to or even lower than that of BA.2.75. Our multiscale investigation provides evidence suggesting that XBB is the first observed SARS-CoV-2 variant to increase its fitness through recombination rather than substitutions.

サルにおけるH5N1不活化全粒子ワクチン接種5年後の肺炎予防効果　　　　　　　　　　　　　　　
仲山 美沙子, 北川 直子, 石垣 宏仁, Nguyen Thanh Cong, 石田 英明, 鈴木 紗織, 小笠原 一誠, 伊藤 靖
日本病理学会会誌, 112, 1, 242, 242, (一社)日本病理学会, 2023年03月
日本語

サルにおけるH5N1不活化全粒子ワクチン接種5年後の肺炎予防効果　　　　　　　　　　　　　　　
仲山 美沙子, 北川 直子, 石垣 宏仁, Nguyen Thanh Cong, 石田 英明, 鈴木 紗織, 小笠原 一誠, 伊藤 靖
日本病理学会会誌, 112, 1, 242, 242, (一社)日本病理学会, 2023年03月
日本語

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

Photochemical Identification of Auxiliary Severe Acute Respiratory Syndrome Coronavirus 2 Host Entry Factors Using μMap
Saori Suzuki, Jacob B. Geri, Steve D. Knutson, Harris Bell-Temin, Tomokazu Tamura, David F. Fernández, Gabrielle H. Lovett, Nicholas A. Till, Brigitte L. Heller, Jinchao Guo, David W. C. MacMillan, Alexander Ploss
Journal of the American Chemical Society, 144, 36, 16604, 16611, American Chemical Society (ACS), 2022年09月14日, ［査読有り］, ［筆頭著者］, ［国際共著］, ［国際誌］
英語, 研究論文（学術雑誌）, Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the infectious agent of the COVID-19 pandemic, remains a global medical problem. Angiotensin-converting enzyme 2 (ACE2) was identified as the primary viral entry receptor, and transmembrane serine protease 2 primes the spike protein for membrane fusion. However, ACE2 expression is generally low and variable across tissues, suggesting that auxiliary receptors facilitate viral entry. Identifying these factors is critical for understanding SARS-Cov-2 pathophysiology and developing new countermeasures. However, profiling host-virus interactomes involves extensive genetic screening or complex computational predictions. Here, we leverage the photocatalytic proximity labeling platform mu Map to rapidly profile the spike interactome in human cells and identify eight novel candidate receptors. We systemically validate their functionality in SARS-CoV-2 pseudoviral uptake assays with both Wuhan and Delta spike variants and show that dual expression of ACE2 with either neuropilin-2, ephrin receptor A7, solute carrier family 6 member 15, or myelin and lymphocyte protein 2 significantly enhances viral uptake. Collectively, our data show that SARS-CoV-2 synergistically engages several host factors for cell entry and establishes mu Map as a powerful tool for rapidly interrogating host-virus interactomes.

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, Elsevier BV, 2022年06月, ［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 formalinand/or b-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 naive 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. (c) 2022 Elsevier Ltd. All rights reserved.

Induction of broadly neutralizing antibodies using a secreted form of the hepatitis C virus E1E2 heterodimer as a vaccine candidate
Ruixue Wang, Saori Suzuki, Johnathan D. Guest, Brigitte Heller, Maricar Almeda, Alexander K. Andrianov, Alexander Marin, Roy A. Mariuzza, Zhen-Yong Keck, Steven K. H. Foung, Abdul S. Yunus, Brian G. Pierce, Eric A. Toth, Alexander Ploss, Thomas R. Fuerst
Proceedings of the National Academy of Sciences, 119, 11, Proceedings of the National Academy of Sciences, 2022年03月15日, ［査読有り］, ［国際共著］, ［国際誌］
研究論文（学術雑誌）, Significance

Hepatitis C virus chronically infects approximately 1% of the world’s population, making an effective vaccine for hepatitis C virus a major unmet public health need. The membrane-associated E1E2 envelope glycoprotein has been used in clinical studies as a vaccine candidate. However, limited neutralization breadth and difficulty in producing large amounts of homogeneous membrane-associated E1E2 have hampered efforts to develop an E1E2-based vaccine. Our previous work described the design and biochemical validation of a native-like soluble secreted form of E1E2 (sE1E2). Here, we describe the immunogenic characterization of the sE1E2 complex. sE1E2 elicited broadly neutralizing antibodies in immunized mice, with increased neutralization breadth relative to the membrane-associated E1E2, thereby validating this platform as a promising model system for vaccine development.

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, Elsevier BV, 2021年06月, ［査読有り］, ［国際共著］, ［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 naive mice and macaques. WPV is thus recommended as an alternative option for seasonal influenza vaccines, especially for children. (c) 2021 Elsevier Ltd. All rights reserved.

SARS-CoV-2 requires cholesterol for viral entry and pathological syncytia formation
David W Sanders, Chanelle C Jumper, Paul J Ackerman, Dan Bracha, Anita Donlic, Hahn Kim, Devin Kenney, Ivan Castello-Serrano, Saori Suzuki, Tomokazu Tamura, Alexander H Tavares, Mohsan Saeed, Alex S Holehouse, Alexander Ploss, Ilya Levental, Florian Douam, Robert F Padera, Bruce D Levy, Clifford P Brangwynne
eLife, 10, eLife Sciences Publications, Ltd, 2021年04月23日, ［査読有り］, ［国際共著］, ［国際誌］
英語, 研究論文（学術雑誌）, Many enveloped viruses induce multinucleated cells (syncytia), reflective of membrane fusion events caused by the same machinery that underlies viral entry. These syncytia are thought to facilitate replication and evasion of the host immune response. Here, we report that co-culture of human cells expressing the receptor ACE2 with cells expressing SARS-CoV-2 spike, results in synapse-like intercellular contacts that initiate cell-cell fusion, producing syncytia resembling those we identify in lungs of COVID-19 patients. To assess the mechanism of spike/ACE2-driven membrane fusion, we developed a microscopy-based, cell-cell fusion assay to screen ~6000 drugs and >30 spike variants. Together with quantitative cell biology approaches, the screen reveals an essential role for biophysical aspects of the membrane, particularly cholesterol-rich regions, in spike-mediated fusion, which extends to replication-competent SARS-CoV-2 isolates. Our findings potentially provide a molecular basis for positive outcomes reported in COVID-19 patients taking statins and suggest new strategies for therapeutics targeting the membrane of SARS-CoV-2 and other fusogenic viruses.

Efficacy of a Cap-Dependent Endonuclease Inhibitor and Neuraminidase Inhibitors against H7N9 Highly Pathogenic Avian Influenza Virus Causing Severe Viral Pneumonia in Cynomolgus Macaques
Saori Suzuki, Cong Thanh Nguyen, Ayako Ogata-Nakahara, Akihiro Shibata, Hiroyuki Osaka, Hirohito Ishigaki, Masatoshi Okamatsu, Yoshihiro Sakoda, Hiroshi Kida, Kazumasa Ogasawara, Yasushi Itoh
Antimicrobial Agents and Chemotherapy, 65, 3, American Society for Microbiology, 2021年02月17日, ［査読有り］, ［筆頭著者］, ［国際誌］
英語, 研究論文（学術雑誌）, H7N9 highly pathogenic avian influenza virus (HPAIV) infection in a human was first reported in 2017. A/duck/Japan/AQ-HE29-22/2017 (H7N9) (Dk/HE29-22), found in imported duck meat at an airport in Japan, possesses a hemagglutinin with a multibasic cleavage site, indicating high pathogenicity in chickens, as in the case of other H7 HPAIVs.

Immune profiling of influenza‐specific B‐ and T‐cell responses in macaques using flow cytometry‐based assays
Marios Koutsakos, Toshiki Sekiya, Brendon Y Chua, Thi Hoang Oanh Nguyen, Adam K Wheatley, Jennifer A Juno, Marumi Ohno, Naoki Nomura, Yuki Ohara, Tomohiro Nishimura, Masafumi Endo, Saori Suzuki, Hirohito Ishigaki, Misako Nakayama, Cong T Nguyen, Yasushi Itoh, Masashi Shingai, Kazumasa Ogasawara, Yoichiro Kino, Stephen J Kent, David C Jackson, Lorena E Brown, Hiroshi Kida, Katherine Kedzierska
Immunology & Cell Biology, 99, 1, 97, 106, Wiley, 2021年01月, ［査読有り］, ［国際共著］, ［国際誌］
研究論文（学術雑誌）

H7N9高病原性鳥インフルエンザウイルスに対するバロキサビルの有効性評価および免疫反応の解析　　　　　　　　　　　　　　　
鈴木 紗織, Nguyen Thanh Cong, 緒方 綾子[中原], 石垣 宏仁, 仲山 美沙子, 伊藤 靖
日本獣医学会学術集会講演要旨集, 163回, 231, 231, (公社)日本獣医学会, 2020年10月
日本語

Animal Models Used in Hepatitis C Virus Research
Keith A. Berggren, Saori Suzuki, Alexander Ploss
International Journal of Molecular Sciences, 21, 11, 3869, 3869, MDPI AG, 2020年05月29日, ［査読有り］, ［筆頭著者］, ［国際共著］, ［国際誌］
研究論文（学術雑誌）, The narrow range of species permissive to infection by hepatitis C virus (HCV) presents a unique challenge to the development of useful animal models for studying HCV, as well as host immune responses and development of chronic infection and disease. Following earlier studies in chimpanzees, several unique approaches have been pursued to develop useful animal models for research while avoiding the important ethical concerns and costs inherent in research with chimpanzees. Genetically related hepatotropic viruses that infect animals are being used as surrogates for HCV in research studies; chimeras of these surrogate viruses harboring specific regions of the HCV genome are being developed to improve their utility for vaccine testing. Concurrently, genetically humanized mice are being developed and continually advanced using human factors known to be involved in virus entry and replication. Further, xenotransplantation of human hepatocytes into mice allows for the direct study of HCV infection in human liver tissue in a small animal model. The current advances in each of these approaches are discussed in the present review.

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

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

カニクイザルを用いたH7N9亜型高病原性鳥インフルエンザウイルスの病原性解析　　　　　　　　　　　　　　　
緒方 綾子, 鈴木 紗織, グエン・タンコン, 伊藤 靖, 石垣 宏仁, 仲山 美沙子, 小笠原 一誠
日本病理学会会誌, 109, 1, 506, 506, (一社)日本病理学会, 2020年03月
日本語

カニクイザルを用いたH7N9亜型高病原性鳥インフルエンザウイルスの病原性解析　　　　　　　　　　　　　　　
緒方 綾子, 鈴木 紗織, グエン・タンコン, 伊藤 靖, 石垣 宏仁, 仲山 美沙子, 小笠原 一誠
日本病理学会会誌, 109, 1, 506, 506, (一社)日本病理学会, 2020年03月
日本語

カニクイザルを用いたH5N6亜型高病原性鳥インフルエンザウイルスによる重症肺炎の解析　　　　　　　　　　　　　　　
伊藤 靖, Nguyen Thanh Cong, 鈴木 紗織, 石垣 宏仁, 仲山 美沙子, 小笠原 一誠
日本病理学会会誌, 108, 1, 285, 285, (一社)日本病理学会, 2019年04月
日本語

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04月, ［査読有り］, ［国際共著］, ［国際誌］
英語, 研究論文（学術雑誌）

Induction of humoural and cellular immunity by immunisation with HCV particle vaccine in a non-human primate model.
Hiroshi Yokokawa, Atsunori Higashino, Saori Suzuki, Masaki Moriyama, Noriko Nakamura, Tomohiko Suzuki, Ryosuke Suzuki, Koji Ishii, Kouji Kobiyama, Ken J Ishii, Takaji Wakita, Hirofumi Akari, Takanobu Kato
Gut, 67, 2, 372, 379, 2018年02月, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, OBJECTIVE: Although HCV is a major cause of chronic liver disease worldwide, there is currently no prophylactic vaccine for this virus. Thus, the development of an HCV vaccine that can induce both humoural and cellular immunity is urgently needed. To create an effective HCV vaccine, we evaluated neutralising antibody induction and cellular immune responses following the immunisation of a non-human primate model with cell culture-generated HCV (HCVcc). DESIGN: To accomplish this, 10 common marmosets were immunised with purified, inactivated HCVcc in combination with two different adjuvants: the classically used aluminum hydroxide (Alum) and the recently established adjuvant: CpG oligodeoxynucleotide (ODN) wrapped by schizophyllan (K3-SPG). RESULTS: The coadministration of HCVcc with K3-SPG efficiently induced immune responses against HCV, as demonstrated by the production of antibodies with specific neutralising activity against chimaeric HCVcc with structural proteins from multiple HCV genotypes (1a, 1b, 2a and 3a). The induction of cellular immunity was also demonstrated by the production of interferon-γ mRNA in spleen cells following stimulation with the HCV core protein. These changes were not observed following immunisation with HCVcc/Alum preparation. No vaccination-related abnormalities were detected in any of the immunised animals. CONCLUSIONS: The current preclinical study demonstrated that a vaccine included both HCVcc and K3-SPG induced humoural and cellular immunity in marmosets. Vaccination with this combination resulted in the production of antibodies exhibiting cross-neutralising activity against multiple HCV genotypes. Based on these findings, the vaccine created in this study represents a promising, potent and safe prophylactic option against HCV.

Persistent replication of a hepatitis C virus genotype 1b-based chimeric clone carrying E1, E2 and p6 regions from GB virus B in a New World monkey
Saori Suzuki, Ken-ichi Mori, Atsunori Higashino, Yuki Iwasaki, Yasuhiro Yasutomi, Noboru Maki, Hirofumi Akari
MICROBIOLOGY AND IMMUNOLOGY, 60, 1, 26, 34, WILEY-BLACKWELL, 2016年01月, ［査読有り］, ［筆頭著者］, ［国内誌］
英語, 研究論文（学術雑誌）, The development of effective hepatitis C virus (HCV) vaccines is essential for the prevention of further HCV dissemination, especially in developing countries. Therefore the aim of this study is to establish a feasible and immunocompetent surrogate animal model of HCV infection that will help in evaluation of the protective efficacy of newly developing HCV vaccine candidates. To circumvent the narrow host range of HCV, an HCV genotype 1b-based chimeric clone carrying E1, E2 and p6 regions from GB virus B (GBV-B), which is closely related to HCV, was generated. The chimera between HCV and GBV-B, named HCV/G, replicated more efficiently as compared with the HCV clone in primary marmoset hepatocytes. Furthermore, it was found that the chimera persistently replicated in a tamarin for more than 2 years after intrahepatic inoculation of the chimeric RNA. Although relatively low (<200 copies/mL), the viral RNA loads in plasma were detectable intermittently during the observation period. Of note, the chimeric RNA was found in the pellet fraction obtained by ultracentrifugation of the plasma at 73 weeks, indicating production of the chimeric virus. Our results will help establish a novel non-human primate model for HCV infection on the basis of the HCV/G chimera in the major framework of the HCV genome.

Increased expression of the regulatory T cell-associated marker CTLA-4 in bovine leukemia virus infection
Saori Suzuki, Satoru Konnai, Tomohiro Okagawa, Ryoyo Ikebuchi, Asami Nishimori, Junko Kohara, Claro N. Mingala, Shiro Murata, Kazuhiko Ohashi
VETERINARY IMMUNOLOGY AND IMMUNOPATHOLOGY, 163, 3-4, 115, 124, ELSEVIER SCIENCE BV, 2015年02月, ［査読有り］, ［筆頭著者］, ［国際共著］, ［国際誌］
英語, 研究論文（学術雑誌）, Regulatory T cells (Tregs) play a critical role in the maintenance of the host's immune system. Tregs, particularly CD4(+)CD25(+)Foxp3(+) T cells, have been reported to be involved in the immune evasion mechanism of tumors and several pathogens that cause chronic infections. Recent studies showed that a Treg-associated marker, cytotoxic T-lymphocyte antigen 4 (CTLA-4), is closely associated with the progression of several diseases. We recently reported that the proportion of Foxp3(+)CD4(+) cells was positively correlated with the number of lymphocytes, virus titer, and virus load but inversely correlated with IFN-gamma expression in cattle infected with bovine leukemia virus (BLV), which causes chronic infection and lymphoma in its host. Here the kinetics of CfLA-4(+) cells were analyzed in BLV-infected cattle. CTLA-4 mRNA was predominantly expressed in CD4+ T cells in BLV-infected cattle, and the expression was positively correlated with Foxp3 mRNA expression. To test for differences in the protein expression level of CTLA-4, we measured the proportion of CTLA-4-expressing cells by flow cytometry. In cattle with persistent lymphocytosis (PL), mean fluorescence intensities (MFIs) of CTLA-4 on CD4(+) and CD25(+) T cells were significantly increased compared with that in control and aleukemic (AL) cattle. The percentage of CTLA-4(+) cells in the CD4(+) T cell subpopulation was positively correlated with TGF-beta mRNA expression, suggesting that CD4(+)CTLA-4(+) T cells have a potentially immunosuppressive function in BLV infection. In the limited number of cattle that were tested, the anti-CTLA-4 antibody enhanced the expression of CD69, IL-2, and IFN-gamma mRNA in anti-programmed death ligand 1 (PD-L1) antibody-treated peripheral blood mononuclear cells from BLV-infected cattle. Together with previous findings, the present results indicate that Tregs may be involved in the inhibition of T cell function during BLV infection. (C) 2014 Elsevier B.V. All rights reserved.

New Type of Sendai Virus Vector Provides Transgene-Free iPS Cells Derived from Chimpanzee Blood
Yasumitsu Fujie, Noemi Fusaki, Tomohiko Katayama, Makoto Hamasaki, Yumi Soejima, Minami Soga, Hiroshi Ban, Mamoru Hasegawa, Satoshi Yamashita, Shigemi Kimura, Saori Suzuki, Tetsuro Matsuzawa, Hirofumi Akari, Takumi Era
PLOS ONE, 9, 12, e113052, PUBLIC LIBRARY SCIENCE, 2014年12月, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, Induced pluripotent stem cells (iPSCs) are potentially valuable cell sources for disease models and future therapeutic applications; however, inefficient generation and the presence of integrated transgenes remain as problems limiting their current use. Here, we developed a new Sendai virus vector, TS12KOS, which has improved efficiency, does not integrate into the cellular DNA, and can be easily eliminated. TS12KOS carries KLF4, OCT3/4, and SOX2 in a single vector and can easily generate iPSCs from human blood cells. Using TS12KOS, we established iPSC lines from chimpanzee blood, and used DNA array analysis to show that the global gene-expression pattern of chimpanzee iPSCs is similar to those of human embryonic stem cell and iPSC lines. These results demonstrated that our new vector is useful for generating iPSCs from the blood cells of both human and chimpanzee. In addition, the chimpanzee iPSCs are expected to facilitate unique studies into human physiology and disease.

HIV-1感染霊長類モデルの樹立に向けた、 Macaca属サルにおけるTRIM5遺伝子の多様性に関する研究　　　　　　　　　　　　　　　
齊藤暁, 河野健, 川本芳, 東濃篤徳, 鈴木紗織, 吉田友教, 鳥居隆三, 保富康宏, 塩田達雄, 中山英美, 明里宏文
日本エイズ学会誌, 16, 1, 28, 36, 2014年, ［招待有り］
日本語, 研究論文（学術雑誌）

Expression analysis of Foxp3 in T cells from bovine leukemia virus infected cattle
Saori Suzuki, Satoru Konnai, Tomohiro Okagawa, Ryoyo Ikebuchi, Tatsuya Shirai, Yuji Sunden, Claro N. Mingala, Shiro Murata, Kazuhiko Ohashi
Microbiology and Immunology, 57, 8, 600, 604, 2013年08月, ［査読有り］, ［筆頭著者］, ［国際共著］, ［国内誌］
英語, 研究論文（学術雑誌）, In the present study, we monitored Foxp3+ T cells in bovine leukemia virus (BLV)-infected cattle. By flow cytometric analysis, the proportion of Foxp3+CD4+ cells from persistent lymphocytotic cattle was significantly increased compared to control and AL cattle. Interestingly, the proportion of Foxp3+CD4+ cells correlated positively with the increased number of lymphocytes, virus titer and virus load, whereas it inversely correlated with IFN-γ mRNA expression, suggesting that Foxp3+CD4+ T cells in cattle have a potentially immunosuppressive function. Further studies are necessary to elucidate the detailed mechanism behind the increased Treg during BLV infection. © 2013 The Societies and Wiley Publishing Asia Pty Ltd.

Efficient in vivo depletion of CD8(+) T lymphocytes in common marmosets by novel CD8 monoclonal antibody administration
Tomoyuki Yoshida, Saori Suzuki, Yuki Iwasaki, Akihisa Kaneko, Akatsuki Saito, Yuki Enomoto, Atsunori Higashino, Akino Watanabe, Juri Suzuki, Kenichi Inoue, Teiko Kuroda, Masahiko Takada, Ryoji Ito, Mamoru Ito, Hirofumi Akari
IMMUNOLOGY LETTERS, 154, 1-2, 12, 17, ELSEVIER SCIENCE BV, 2013年07月, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, In order to directly demonstrate the roles of CD8(+) T lymphocytes in non-human primates, in vivo depletion of the CD8(+) T cells by administration of a CD8-specific monoclonal antibody (mAb) is one of the crucial techniques. Recently, the common marmoset (Callithrix jacchus), which is classified as a New World monkey, has been shown useful as an experimental animal model for various human diseases such as multiple sclerosis, Parkinson's disease and a number of infectious diseases. Here we show that an anti-marmoset CD8 mAb 6F10, which we have recently established, efficiently depletes the marmoset CD8+ T lymphocytes in vivo, i.e., the administration of 6F10 induces drastic and specific reduction in the ratio of the CD8(+) T cell subset for at least three weeks or longer. Our finding will help understand the pivotal role of CD8(+) T cells in vivo in the control of human diseases. (C) 2013 Elsevier B.V. All rights reserved.

Expression analysis of Foxp3 in T-cells from bovine leukemia virus infected cattle.
Suzuki S, Konnai S, Okagawa T, Ikebuchi R, Shirai T, Sunden Y, Mingala CN, Murata S, Ohashi K
Microbiology and immunology, 2013年06月, ［査読有り］, ［筆頭著者］, ［国際共著］, ［国内誌］

Enhanced expression of LAG-3 on lymphocyte subpopulations from persistently lymphocytotic cattle infected with bovine leukemia virus
Satoru Konnai, Saori Suzuki, Tatsuya Shirai, Ryoyo Ikebuchi, Tomohiro Okagawa, Yuji Sunden, Claro N. Mingala, Misao Onuma, Shiro Murata, Kazuhiko Ohashi
COMPARATIVE IMMUNOLOGY MICROBIOLOGY AND INFECTIOUS DISEASES, 36, 1, 63, 69, ELSEVIER SCI LTD, 2013年01月, ［査読有り］, ［筆頭著者］, ［国際共著］, ［国際誌］
英語, 研究論文（学術雑誌）, An immunoinhibitory receptor, lymphocyte activation gene-3 (LAG-3), which is mainly expressed in T-cells, is involved in the immune evasion of several pathogens causing chronic infections and tumors. However, unlike human or mouse LAG-3, no functional analysis of LAG-3 has been reported in domestic animals. Thus, in this study, bovine LAG-3 expression was analyzed in bovine leukemia virus (BLV)-infected cattle. In persistent lymphocytotic (PL) cattle, the numbers of LAG-3(+)CD4(+) cells and LAG-3(+)CD8(+) cells were conserved whilst the number of MHC class II+ cells was remarkably higher than in the control animals. In contrast, the mean fluorescence intensity (MFI) for LAG-3 on PBMCs from PL cattle was significantly increased compared to control and asymptomatic (AL) cattle. Specifically, the LAG-3 expression level was significantly increased in both CD4(+) and CD8(+) T cells from PL cattle. LAG-3 expression correlated positively with increased numbers of lymphocytes and MHC class II+ cells in infected animals. Preliminary results from PD-L1 and LAG-3 blockade assay revealed that IFN-gamma and IL-2 expressions were significantly up-regulated by addition of anti- PD-L1 and LAG-3 antibodies in PBMCs from PL cattle. These findings suggest that LAG-3 might be involved in the inhibition of T-cell function through its binding and signaling on MHC class II molecule during BLV infection. (c) 2012 Elsevier Ltd. All rights reserved.

Transcriptional profiling of inflammatory cytokine genes in African, buffaloes (Syncerus caffer) infected with Theileria parva
Tomohiro Okagawa, Satoru Konnai, Hirohisa Mekata, Naftaly Githaka, Saori Suzuki, Edward Kariuki, Francis Gakuya, Esther Kanduma, Tatsuya Shirai, Ryoyo Ikebuchi, Yoshinori Ikenaka, Mayumi Ishizuka, Shiro Murata, Kazuhiko Ohashi
VETERINARY IMMUNOLOGY AND IMMUNOPATHOLOGY, 148, 3-4, 373, 379, ELSEVIER SCIENCE BV, 2012年08月, ［査読有り］, ［国際共著］, ［国際誌］
英語, 研究論文（学術雑誌）, Theileria parva (T. parva) causes East Coast fever (ECF), which is of huge economic importance to Eastern and Southern African countries. In a previous bovine model, inflammatory cytokines were closely associated with disease progression in animals experimentally infected with T. parva. The African Cape buffalo (Syncerus coffer), the natural reservoir for T. parva, is completely resistant to ECF despite a persistently high parasitaemia following infection with T. parva. Characterizing basic immunological interactions in the host is critical to understanding the mechanism underlying disease resistance in the African Cape buffalo. In this study, the expression level of several cytokines was analyzed in T. parva-infected buffaloes. There were no significant differences in the expression profiles of inflammatory cytokines between the infected and uninfected animals despite a remarkably high parasitaemia in the former. However, the expression level of IL-10 was significantly upregulated in the infected animals. These results indicate a correlation between diminished inflammatory cytokines response and disease resistance in the buffalo. (c) 2012 Elsevier B.V. All rights reserved.

Increased bovine Tim-3 and its ligand expressions during bovine leukemia virus infection
Tomohiro Okagawa, Satoru Konnai, Ryoyo Ikebuchi, Saori Suzuki, Tatsuya Shirai, Yuji Sunden, Misao Onuma, Shiro Murata, Kazuhiko Ohashi
VETERINARY RESEARCH, 43, 45, BIOMED CENTRAL LTD, 2012年05月, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, The immunoinhibitory receptor T cell immunoglobulin domain and mucin domain-3 (Tim-3) and its ligand, galectin-9 (Gal-9), are involved in the immune evasion mechanisms for several pathogens causing chronic infections. However, there is no report concerning the role of Tim-3 in diseases of domestic animals. In this study, cDNA encoding for bovine Tim-3 and Gal-9 were cloned and sequenced, and their expression and role in immune reactivation were analyzed in bovine leukemia virus (BLV)-infected cattle. Predicted amino acid sequences of Tim-3 and Gal-9 shared high homologies with human and mouse homologues. Functional domains, including tyrosine kinase phosphorylation motif in the intracellular domain of Tim-3 were highly conserved among cattle and other species. Quantitative real-time PCR analysis showed that bovine Tim-3 mRNA is mainly expressed in T cells such as CD4(+) and CD8(+) cells, while Gal-9 mRNA is mainly expressed in monocyte and T cells. Tim-3 mRNA expression in CD4(+) and CD8(+) cells was upregulated during disease progression of BLV infection. Interestingly, expression levels for Tim-3 and Gal-9 correlated positively with viral load in infected cattle. Furthermore, Tim-3 expression level closely correlated with up-regulation of IL-10 in infected cattle. The expression of IFN-gamma and IL-2 mRNA was upregulated when PBMC from BLV-infected cattle were cultured with Cos-7 cells expressing Tim-3 to inhibit the Tim-3/Gal-9 pathway. Moreover, combined blockade of the Tim-3/Gal-9 and PD-1/PD-L1 pathways significantly promoted IFN-gamma mRNA expression compared with blockade of the PD-1/PD-L1 pathway alone. These results suggest that Tim-3 is involved in the suppression of T cell function during BLV infection.

Molecular cloning and characterization of Th1 and Th2 cytokines of African buffalo (Syncerus caffer)
S. Suzuki, S. Konnai, T. Okagawa, N. W. Githaka, E. Kariuki, F. Gakuya, E. Kanduma, T. Shirai, R. Ikebuchi, Y. Ikenaka, M. Ishizuka, S. Murata, K. Ohashi
INTERNATIONAL JOURNAL OF IMMUNOGENETICS, 39, 2, 170, 182, WILEY-BLACKWELL, 2012年04月, ［査読有り］, ［筆頭著者］, ［国際共著］, ［国際誌］
英語, 研究論文（学術雑誌）, The African buffalo (Syncerus caffer) has been implicated as the reservoir of several bovine infectious agents. However, there is insufficient information on the protective immune responses in the African buffalo, particularly in infected animals. In this study, we analysed Th1 cytokines IL-2 and IFN-?, and Th2 cytokines IL-4 and IL-10. The cloned cDNA of IL-2, IL-4, IL-10 and IFN-? contained an open reading frame of 468, 501, 408 and 540 nucleotides, encoding polypeptides of 155, 166, 135 and 179 amino acids, respectively. Nucleotide sequence homology of IL-2, IFN-? and IL-4 was more than 98% between the African buffalo and cattle, which resulted in identical polypeptides. Meanwhile, IL-10 gene of African buffalo and cattle had 95% homology in nucleotide sequence, corresponding to thirteen amino acid residues substitution. Cysteine residues and potential glycosylation sites were conserved within the family Bovinae. Phylogenetic analyses including cytokines of the African buffalo placed them within a cluster comprised mainly of species belonging to the order Artiodactyla, including cattle, water buffalo, sheep, goat, pig and artiodactyl wildlife. A deeper understanding of the structure of these cytokines will shed light on their protective role in the disease-resistant African buffalo in comparison with other closely related species.

Molecular cloning of bovine lymphocyte activation gene-3 and its expression characteristics in bovine leukemia virus-infected cattle
Tatsuya Shirai, Satoru Konnai, Ryoyo Ikebuchi, Tomohiro Okagawa, Saori Suzuki, Yuji Sunden, Misao Onuma, Shiro Murata, Kazuhiko Ohashi
VETERINARY IMMUNOLOGY AND IMMUNOPATHOLOGY, 144, 3-4, 462, 467, ELSEVIER SCIENCE BV, 2011年12月, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, Lymphocyte activation gene-3 (LAG-3), a major histocompatibility complex (MHC) class II binding CD4 homologue has recently been shown as one of the mechanisms for down-regulating immune responses during chronic disease progression. For the first time, we cloned LAG-3 from two breeds of cattle (Holstein and Japanese Black), and analyzed its expression levels in cattle infected with bovine leukemia virus (BLV), a chronic viral infection that leads to immuno-suppression. The cloned cDNA of bovine LAG-3 have an open reading frame of 1551 nucleotides, encoding a polypeptide of 515 amino acids in length. Similar to the swine LAG-3, the bovine LAG-3 protein sequence consisted of four extracellular domains, a transmembrane domain and an inhibitory motif, KTGELE. We found that the bovine LAG-3 mRNA transcripts were expressed predominantly on T-cells such as CD4(+) and CD8(+) cells, among peripheral blood mononuclear cells. In subsequent expression analysis, LAG-3 mRNA expression on CD4(+) T-cells from BLV-infected cattle was upregulated compared to that in normal cattle. Comparable results were obtained with CD8(+) T-cells from cattle infected with BLV. We further observed strong upregualtion of MHC class II molecule, the ligand for LAG-3 in BLV-infected cattle. These findings indicate an important role for inhibitory receptor molecules such as LAG-3 in chronic bovine infections and future studies will elucidate the specific role of LAG-3 in bovine diseases. (C) 2011 Elsevier B.V. All rights reserved.

μMap技術を用いたSARS-CoV2のエントリーに関わる宿主補助因子の同定
鈴木紗織, GERI Jacob, GERI Jacob, KNUTSON Steve, BELL-TEMIN Harris, 田村友和, FERNANDEZ David, LOVETT Gabby, LI Beryl, DONG Zhe, TILL Nick, MACMILLAN David W.C., MACMILLAN David W.C., PLOSS Alexander, 日本獣医学会学術集会講演要旨集, 165th (CD-ROM), 2022年

カニクイザルを用いたH5N6亜型高病原性鳥インフルエンザウイルスによる重症肺炎の解析　　　　　　　　　　　　　　　
伊藤 靖, Nguyen Thanh Cong, 鈴木 紗織, 石垣 宏仁, 仲山 美沙子, 小笠原 一誠, 日本病理学会会誌, 108, 1, 285, 285, 2019年04月
(一社)日本病理学会, 日本語

H7N9亜型鳥インフルエンザウイルスのノイラミニダーゼ阻害薬耐性変異とその病原性　　　　　　　　　　　　　　　
鈴木 紗織, 七戸 新太郎, 伊藤 靖, 小笠原 一誠, 日本獣医学会学術集会講演要旨集, 161回, 355, 355, 2018年08月
(公社)日本獣医学会, 日本語

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

カニクイザルでのH7N9鳥インフルエンザHAタンパク質発現組換えワクシニアワクチンの発症防御効果の検討
山地賢三郎, 安井文彦, 伊藤靖, 鈴木紗織, 本田智子, 山本直樹, 真田崇弘, 石垣宏仁, 石井孝司, 小笠原一誠, 小原道法, 日本ワクチン学会学術集会プログラム・抄録集, 22nd, 118, 2018年
日本語

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

ノイラミニダーゼ阻害薬耐性H7N9亜型鳥インフルエンザウイルスの薬剤非投与カニクイザルにおける病原性評価　　　　　　　　　　　　　　　
鈴木 紗織, 七戸 新太郎, 伊藤 靖, 小笠原 一誠, 日本獣医学会学術集会講演要旨集, 160回, 385, 385, 2017年08月
(公社)日本獣医学会, 日本語

持続感染した霊長類肝炎ウイルスの適応変異:ウイルスゲノム変異が及ぼす病態への影響
東濃篤徳, 鈴木紗織, 森健一, 大出裕高, 松岡和弘, 片貝祐子, 岡林佐知, 槇昇, 岩谷靖雅, 杉浦亙, 明里宏文, 霊長類研究, 31, Supplement, 81, 82, 2015年06月20日
日本語

HCVベースのHCV/GBV-Bキメラウイルスはタマリンへ長期感染する　　　　　　　　　　　　　　　
鈴木紗織, 東濃篤徳, 森健一, 片貝祐子, 槇昇, 明里宏文, 第62回日本実験動物学会学術集会, 2015年, ［査読有り］

新世界ザルにおける液性免疫応答の機能低下は霊長類肝炎ウイルスの持続感染に寄与する　　　　　　　　　　　　　　　
鈴木紗織, 東濃篤徳, 森健一, 大出裕高, 松岡和弘, 岩谷靖雅, 杉浦亙, 片貝祐子, 槇昇, 明里宏文, 第31回日本霊長類学会大会, 2015年, ［査読有り］

Genetic and immunological escape in persistent GBV-B infection　　　　　　　　　　　　　　　
Saori Suzuki, Atsunori Higashino, Ken-ichi Mori, Hirotaka Ode, Kazuhiro Matsuoka, Yasumasa Iwatani, Wataru Sugiura, Yuko Katakai, Noboru Maki, Hirofumi Akari, 22nd International Symposium on Hepatitis C Virus and Related Viruses, 2015年, ［査読有り］

慢性GBV-B感染におけるenvelope変異および液性免疫からの逃避　　　　　　　　　　　　　　　
鈴木紗織, 東濃篤徳, 森健一, 大出裕高, 松岡和弘, 岩谷靖雅, 杉浦亙, 片貝祐子, 槇昇, 明里宏文, 第63回日本ウイルス学会学術集会, 2015年, ［査読有り］

霊長類ヘパチウイルス慢性感染後寛解におけるウイルスゲノム変異の意義　　　　　　　　　　　　　　　
東濃篤徳, 鈴木紗織, 大出裕高, 松岡和弘, 森健一, 片貝祐子, 岡林佐知, 槇昇, 岩谷靖雅, 杉浦亙, 明里宏文, 第63回日本ウイルス学会学術集会, 2015年, ［査読有り］

培養細胞由来HCV粒子のマーモセットにおける抗HCV抗体誘導能の検討
横川寛, 中村紀子, 東濃篤徳, 鈴木紗織, 明里宏文, 加藤孝宣, 石井孝司, 脇田隆字, 日本ウイルス学会学術集会プログラム・抄録集, 62nd, 338, 2014年10月31日
日本語

小型霊長類モデルを用いたヘパチウイルスの持続感染における慢性肝炎発症に影響するウイルスゲノム変異解析　　　　　　　　　　　　　　　
東濃篤徳, 鈴木紗織, 齊藤暁, 松岡和弘, 大出裕高, 片貝祐子, 岡林佐知, 森健一, 槇昇, 明里宏文, 第61回日本実験動物学会総会, 2014年, ［査読有り］

Diversity of antiretroviral host factor TRIM5 gene in macaque monkeys　　　　　　　　　　　　　　　
Akatsuki Saito, Emi E Nakayama, Tatsuo Shioda, Tomoyuki Yoshida, Atsunori Higashino, Saori Suzuki, Yoshi Kawamoto, Hirofumi Akari, Cold Spring Harbor meeting on Retroviruses, 2013年, ［査読有り］

急性及び慢性GBV-B感染症における液性免疫の比較解析　　　　　　　　　　　　　　　
鈴木紗織, 東濃篤徳, 森健一, 吉田友教, 片貝祐子, 齊藤暁, 槇昇, 明里宏文, 第61回日本ウイルス学会学術集会, 2013年, ［査読有り］

CCR5指向性を示す新規サル指向性HIV-1はサル個体に持続感染する　　　　　　　　　　　　　　　
齊藤暁, 大附寛幸, 東濃篤徳, 鈴木紗織, 松田健太, 高橋尚史, 松岡佐織, 岩谷靖雅, 杉浦亙, 野間口雅子, 足立昭夫, 保富康宏, 俣野哲朗, 三浦智行, 明里宏文, 第61回日本ウイルス学会学術集会, 2013年, ［査読有り］

新世界ザルにおける持続感染GBV-BのQuasispecies解析　　　　　　　　　　　　　　　
東濃篤徳, 鈴木紗織, 齊藤暁, 片貝祐子, 岡林佐知, 明里宏文, 第61回日本ウイルス学会学術集会, 2013年, ［査読有り］

The delayed humoral immune responses may be associated with the development of chronic GBV-B infection　　　　　　　　　　　　　　　
Saori Suzuki, Atsunori Higashino, Ken-ichi Mori, Yuko Katakai, Akatsuki Saito, Noboru Maki, Hirofumi Akari, 20th International Symposium on Hepatitis C Virus and Related Viruses, 2013年, ［査読有り］

Analysis of dynamics in GB virus B quasispecies in the course of long-term persistent infection and disease progression in marmosets　　　　　　　　　　　　　　　
Atsunori Higashino, Saori Suzuki, Akatsuki Saito, Yuko Katakai, Sachi Okabayashi, Hirofumi Akari, 20th International Symposium on Hepatitis C Virus and Related Viruses, 2013年, ［査読有り］

ウイルスの標的細胞指向性はサル指向性HIV-1の増殖に影響するか？　　　　　　　　　　　　　　　
齊藤暁, 大附寛幸, 東濃篤徳, 鈴木紗織, 松田健太, 高橋尚史, 岩谷靖雅, 杉浦亙, 保富康宏, 俣野哲朗, 三浦智行, 明里宏文, 第156回日本獣医学会学術集会, 2013年, ［査読有り］

GBV-B感染新世界ザルの液性免疫解析　　　　　　　　　　　　　　　
鈴木紗織, 東濃篤徳, 森健一, 吉田友教, 齊藤暁, 槇昇, 明里宏文, 第60回日本実験動物学会総会, 2013年, ［査読有り］

レトロウイルス感受性に影響するマカク属サルTRIM5遺伝子の多様性　　　　　　　　　　　　　　　
齊藤暁, 中山英美, 塩田達雄, 吉田友教, 東濃篤徳, 鈴木紗織, 川本芳, 明里宏文, 第60回日本実験動物学会総会, 2013年, ［査読有り］

霊長類を用いたHCV/ GBV-Bキメラウイルス感染モデル　　　　　　　　　　　　　　　
東濃篤徳, 森健一, 鈴木紗織, 岩崎優紀, 吉田友教, 齊藤暁, 槇昇, 明里宏文, 第60回日本実験動物学会総会, 2013年, ［査読有り］

タマリンを用いたHCV/GBV‐Bキメラウイルス感染モデル
東濃篤徳, 森健一, 鈴木紗織, 岩崎優紀, 吉田友教, 齊藤暁, 槇昇, 明里宏文, 日本ウイルス学会学術集会プログラム・抄録集, 60th, 413, 2012年10月31日, ［査読有り］
日本語

マカク属サルTRIM5遺伝子における種間および種内の多様性
齊藤暁, 河野健, 中山英美, 塩田達雄, 川本芳, 鳥居隆三, 吉田友教, 東濃篤徳, 鈴木紗織, 保富康宏, 明星宏文, 日本ウイルス学会学術集会プログラム・抄録集, 60th, 343, 2012年10月31日, ［査読有り］
日本語

牛白血病ウイルス感染牛における免疫抑制因子Tim‐3/Gal‐9の発現解析および機能解析
岡川朋弘, 今内覚, 池渕良洋, 鈴木紗織, 寸田祐嗣, 村田史郎, 大橋和彦, 日本獣医学会学術集会講演要旨集, 154th, 233, 2012年08月31日
日本語

ウシ難治性疾病に対する多機能型新規治療法の開発　　　　　　　　　　　　　　　
岡川朋弘, 今内覚, 池渕良洋, 鈴木紗織, 村田史郎, 大橋和彦, 動物ワクチン-バイオ医薬品研究会ニュースレター, 5, 21, 23, 2012年06月, ［招待有り］
日本語, 記事・総説・解説・論説等（学術雑誌）

Theileria parva感染牛属のサイトカイン発現比較解析　　　　　　　　　　　　　　　
岡川朋弘, 今内覚, Githaka N, 目堅博久, 鈴木紗織, Kariuki E, Skilton R, 石塚真由美, 村田史郎, 大橋和彦, 獣医寄生虫学会誌, 10, 1-2, 37, 2012年03月
日本語, 速報，短報，研究ノート等（学術雑誌）

牛難治性疾病に対する免疫抑制因子を標的とした治療型ワクチンの開発
今内覚, 池渕良洋, 岡川朋弘, 鈴木紗織, 白井達哉, 村田史郎, 大橋和彦, 日本獣医学会学術集会講演要旨集, 153rd, 117, 2012年03月01日
日本語

An animal model for chimeric virus of hepatitis C virus/GB virus B　　　　　　　　　　　　　　　
Atsunori Higashino, Ken-ichi Mori, Saori Suzuki, Yuki Iwasaki, Tomoyuki Yoshida, Akatsuki Saito, Noboru Maki, Hirofumi Akari, 19th International Symposium on Hepatitis C Virus and Related Viruses, 2012年, ［査読有り］

ウシTim-3の発現・機能解析と新規疾病制御法への応用展開　　　　　　　　　　　　　　　
岡川朋弘, 今内覚, 池渕良洋, 鈴木紗織, 村田史郎, 大橋和彦, 動物ワクチン-バイオ医薬品研究会ニュースレター, 4, 27, 29, 2011年12月, ［招待有り］
日本語, 記事・総説・解説・論説等（学術雑誌）

ウシ免疫抑制受容体Tim‐3の同定と牛白血病ウイルス感染牛における発現解析
岡川朋弘, 今内覚, 池渕良洋, 鈴木紗織, 寸田祐嗣, 村田史郎, 大橋和彦, 日本獣医学会学術集会講演要旨集, 152nd, 224, 2011年08月31日
日本語

牛白血病ウイルス感染牛における制御性T細胞の動態解析
鈴木紗織, 今内覚, 池渕良洋, 岡川朋弘, 寸田祐嗣, 村田史郎, 大橋和彦, 日本獣医学会学術集会講演要旨集, 152nd, 224, 2011年08月31日
日本語

Theileria parva原虫感染牛属のサイトカイン発現比較解析
今内覚, 岡川朋弘, NAFTALY Githaka, 目堅博久, 鈴木紗織, EDWARD Kariuki, ROBERT Skilton, 石塚真由美, 村田史郎, 大橋和彦, 日本獣医学会学術集会講演要旨集, 152nd, 204, 2011年08月31日
日本語

アフリカスイギュウ(Syncerus caffer)のTh1,Th2および炎症性サイトカイン遺伝子の同定
岡川朋弘, 鈴木紗織, 今内覚, GITHAKA Naftali, KARIUKI Edward, GAKUYA Francis, ESTHER Kanduma, 石塚真由美, 村田史郎, 大橋和彦, 日本獣医学会学術集会講演要旨集, 150th, 212, 2010年09月01日
日本語

インフルエンザウイルス感染カニクイザルを用いたサイトカインストーム発生機序の解明
科学研究費助成事業
2018年04月01日 - 2020年03月31日
鈴木 紗織
本研究では、免疫抑制因子および抑制性サイトカインに着目し、インフルエンザの重篤化に関与する寄与因子を同定することを目的としている。 まず2017年に分離された高病原性H7N9鳥インフルエンザウイルスを12頭のカニクイザルに感染させ、抗インフルエンザ薬であるOseltamivir, Zanamivir, Baloxavir及びコントロールとして蒸留水をそれぞれ3頭のカニクイザルに5日間投与した(Baloxavirのみ感染後1日目のみ投与)。感染8日目に安楽殺したが、コントロール群の1頭のみ感染3日目でエンドポイントに達し安楽殺した。 ウイルス量はコントロール群で最も多く検出され、特にエンドポイント個体を重症化個体とした。薬剤投与群ではOseltamivir>Zanamivir>Baloxavirの順に検出されたウイルス量が少なかった。さらにサルの末梢血サンプルを用いて、フローサイトメトリーにて免疫抑制因子の発現を解析したところ、どの群も感染3日目でCD4T細胞が多く動員されていた。またPD-1、CTLA-4、TIGIT、LAG-3は感染前や非感染群と比べて、感染後に発現の増加が見られ、コントロール群において顕著だった。さらに、CD8T細胞中のPD-1及びTIGITに関して重症化個体における発現量が特に高く、薬剤投与群はこれらの免疫抑制因子は感染8日目には感染前と同程度の発現まで低下していた。サイトカイン産生量では、重症化個体のIFNa, IFNb, IFNg及びIL-6の産生が他個体より多かったが、IL-10に関しては同程度だった。この結果より、PD-1, TIGITがIFNsやIL-6といった炎症性サイトカインと相互作用して重篤化に関与していることが示唆された。現在は研究中断中であり、再開後詳細に解析する予定である。
日本学術振興会, 若手研究, 滋賀医科大学, 18K15117

新規ＨＣＶ／ＧＢＶ－Ｂキメラウイルスによる革新的Ｃ型肝炎霊長類モデルの構築
科学研究費助成事業
2013年04月01日 - 2016年03月31日
鈴木 紗織
C 型肝炎のワクチン開発に必要な安全性・有効性評価や C 型肝炎の病態解析には大きな障害が存在する。その障害の一つとして適切な感染モデルがないことが挙げられる。本研究ではこのブレイクスルーを目標として、C型肝炎ウイルス（HCV）と新世界ザルに肝炎を引き起こすGBウイルスB（GBV－B）に着目し新世界ザル指向性 HCV/GBV-B キメラウイルスの構築を目指した。
昨年度に作製したGBV-B/HCVキメラウイルスがマーモセットに感染はしたものの持続感染は示さなかったことから、持続感染へのメカニズムをGBV－B感染サルを用いて解析した。すると急性感染後にウイルスが排除される場合は、NK細胞のうち細胞障害活性マーカー（CD335、CD159a）を表出しているNK細胞（CD3-CD56dimCD16+）の割合がウイルスの動態に少し遅れて平行に増減していることがわかった。一方で慢性感染を起こしたGBV－B感染タマリンの慢性期を解析すると、GBV-B非感染サルと同程度の低い割合を示していた。このことから、GBV－B感染において急性期後にクリアランスされるにはNK細胞の細胞障害活性が重要であることが示唆された。さらに液性免疫の視点から解析すると、慢性感染個体では血漿中の抗core、E2、NS3抗体価の誘導の遅延および低い抗体価が検出された。E2に関しては中和抗体のターゲットになるため、抗E2抗体に中和活性がある可能性が考えられる。
以上の結果から、持続感染にはNK細胞の細胞傷害活性と抗体誘導が重要な因子であることが示唆された。今後は、昨年度までに確立したCD8 depletionの方法を用いて、細胞傷害活性を弱めたサルにキメラウイルスを接種し、より効率よくウイルスが増殖できるモデルに改良したい。
日本学術振興会, 特別研究員奨励費, 京都大学, 13J01032