New rotavirus vaccine platforms using reverse genetics systems

• Project/Area Number: 18H02663
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Review Section: Basic Section 49060:Virology-related
• Research Institution: Osaka University
• Principal Investigator: Kobayashi Takeshi 大阪大学, 微生物病研究所, 教授 (90324847)
• Project Period (FY): 2018-04-01 – 2021-03-31
• Project Status: Completed (Fiscal Year 2020)
• Budget Amount: ¥17,160,000 (Direct Cost: ¥13,200,000、Indirect Cost: ¥3,960,000); Fiscal Year 2020: ¥5,720,000 (Direct Cost: ¥4,400,000、Indirect Cost: ¥1,320,000); Fiscal Year 2019: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000); Fiscal Year 2018: ¥6,240,000 (Direct Cost: ¥4,800,000、Indirect Cost: ¥1,440,000)
• Keywords: ワクチン / ロタウイルス / リバースジェネティクス系

Outline of Final Research Achievements

Rotaviruses are a major cause of severe gastroenteritis in infants and young children worldwide and are responsible for approximately 215,000 deaths annually, particularly in developing countries. The objective of this research is to understand how rotaviruses replicate in vitro and cause diseases in vivo using plasmid-based rotavirus reverse genetics systems. We established a plasmid-based reverse genetics system for G4P[8] human rotavirus strain Odelia. Using reverse genetics systems, we recovered a panel of monoreassortant viruses harboring one gene segment from strain Odelia within the simian rotavirus strain SA11 genetic backbone. We also tried to develop a reverse genetics system for murine rotavirus strain EW. Reassortant viruses between strains SA11 and EW generated by the rescue system enabled study of the biological functions of viral gene segments. These systems will provide insight into the molecular mechanisms underlying rotavirus replication and pathogenesis.

Academic Significance and Societal Importance of the Research Achievements

現在、使用されているロタウイルスワクチンにおいては、発展途上国における不十分な防御効果、高額な費用、副反応（腸重積症）等の問題があげられており、次世代ワクチンの開発研究は重要な課題である。これまで有用なリバースジェネティクス系の開発の遅れからロタウイルスの複製機構ならびに病態発現機序の解明は進んでいなかった。本研究成果で開発されたヒトおよびマウスロタウイルス株のリバースジェネティクス系やレポーター遺伝子発現ロタウイルスを用いた研究は新規ワクチン開発や感染伝播、病態発現機序の理解につながると期待される。

Research Products

• [Journal Article] Reverse Genetics Approach for Developing Rotavirus Vaccine Candidates Carrying VP4 and VP7 Genes Cloned from Clinical Isolates of Human Rotavirus (2021)
  • Author(s): Kanai Y, Onishi M, Kawagishi T, Pimfhun P, Nurdin J, Nouda R, Yamasaki M, Tina L, Pattara K, Okitsu S, Hayakawa S, Ebina H, Ushijima H, Kobayashi T
  • Journal Title: Journal of Virology Volume: 95 Issue: 2
  • DOI: 10.1128/jvi.01374-20
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Generation of recombinant rotaviruses encoding a split NanoLuc peptide tag (2021)
  • Author(s): Pimfhun P, Kanai Y, Kawagishi T, Nouda R, Nurdin J, Yamasaki M, Nomura K, Tina L, Kobayashi T
  • Journal Title: Biochemical and Biophysical Research Communications Volume: 534 Pages: 740-746
  • DOI: 10.1016/j.bbrc.2020.11.007
  • Peer Reviewed
• [Journal Article] Construction and characterization of an infectious clone generated from Chikungunya virus SL11131 strain (2021)
  • Author(s): Suzuki Y, Tanaka A, Maeda Y, Emi A, Fujioka Y, Sakaguchi S, SG Vasudevan, Kobayashi T , CK Lim, Takasaki T, H Wu, Nakano T
  • Journal Title: Virology Volume: 552 Pages: 52-62
  • DOI: 10.1016/j.virol.2020.09.006
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Generation of Genetically RGD-Modified Oncolytic Reovirus That Enhances JAM-A-Independent Infection of Tumor Cells (2020)
  • Author(s): Kawagishi T, Kanai Y, Nouda R, Fukui I, Nurdin JA, Matsuura Y, Kobayashi T
  • Journal Title: Journal of Virology Volume: 94 Issue: 23
  • DOI: 10.1128/jvi.01703-20
  • Peer Reviewed
• [Journal Article] Reverse Genetics System for a Human Group A Rotavirus. (2020)
  • Author(s): Kawagishi T, Nurdin J, Onishi M, Nouda R, Kanai Y, Tajima T, Ushijima H, Kobayashi T
  • Journal Title: J Virol Volume: 94 Issue: 2
  • DOI: 10.1128/jvi.00963-19
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Cell-cell fusion induced by reovirus FAST proteins enhances replication and pathogenicity of non-enveloped dsRNA viruses. (2019)
  • Author(s): 1.Kanai Y, Kawagishi T, Sakai Y, Nouda R, Shimojima M, Saijo M, Matsuura Y, Kobayashi T.
  • Journal Title: PLoS Pathog. Volume: 15 Issue: 4 Pages: e1007675-e1007675
  • DOI: 10.1371/journal.ppat.1007675
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] In Vivo Live Imaging of Oncolytic Mammalian Orthoreovirus Expressing NanoLuc Luciferase in Tumor Xenograft Mice (2019)
  • Author(s): Kanai Yuta、Kawagishi Takahiro、Matsuura Yoshiharu、Kobayashi Takeshi
  • Journal Title: Journal of Virology Volume: 93 Issue: 14
  • DOI: 10.1128/jvi.00401-19
  • Peer Reviewed / Open Access
• [Journal Article] Development of Stable Rotavirus Reporter Expression Systems (2019)
  • Author(s): Kanai Y, Kawagishi T, Nouda R, Onishi M, Pannacha P, Nurdin JA, Nomura K, Matsuura Y, Kobayashi T.
  • Journal Title: Journal of Virology Volume: 93 Issue: 4
  • DOI: 10.1128/jvi.01774-18
  • Peer Reviewed
• [Presentation] ロタウイルス感染における多様な細胞表面糖鎖レセプターの意義 (2020)
  • Author(s): 山﨑 萌子、金井 祐太、浜島 りな、南 昌平、Pannacha Pimfhun、納田 遼太郎、Jeffry Nurdin、Tina Lusiany、松本 真依、大西 未紗、小林 剛
  • Organizer: 第163回日本獣医学会学術集会
• [Presentation] Reverse Genetics System for a Human Group A Rotavirus (2020)
  • Author(s): Kawagishi T, Nurdin A, Onishi M, Nouda R, Kanai Y, Tajima T, Ushijima H, Kobayashi T.
  • Organizer: U.S.-Japan Cooperative Medical Sciences Program's 22nd International Conference on Emerging Infectious Diseases in the Pacific Rim
  • Int'l Joint Research
• [Presentation] The Application of Reverse Genetics Systems in Studies of dsRNA Virus Replication and Pathogenesis (2019)
  • Author(s): Kobayashi T.
  • Organizer: Gordon Research Conference Viruses and Cells
  • Int'l Joint Research / Invited
• [Presentation] Reverse Genetic System for Human Rotavirus A Odelia strain (2019)
  • Author(s): Nurdin J, Kawagishi T, Onishi M, Kanai Y, Tajima T, Ushijima H, Kobayashi T.
  • Organizer: The 18th Awaji International Forum on Infection and Immunity
• [Presentation] ロタウイルス NSP3タンパク質のeIF4G結合領域変異がウイルス複製に及ぼす影響 (2019)
  • Author(s): 納田遼太郎, 金井祐太, 川岸崇裕, Pannacha Pimfhun, 小林剛
  • Organizer: 第67回日本ウイルス学会学術集会
• [Presentation] Reverse Genetic System for Human Rotavirus A Odelia strain (2019)
  • Author(s): Nurdin J, Kawagishi T, Onishi M, Kanai Y, Tajima T, Ushijima H, Kobayashi T.
  • Organizer: 第67回日本ウイルス学会学術集会
• [Presentation] ロタウイルスベクターの開発 (2019)
  • Author(s): 金井祐太, 川岸崇裕, Pannacha Pimfhun , 納田遼太郎, 大西未沙, Nurdin Jeffry, 野村圭一郎, Tina Luciany, 山﨑萌子, 小林剛
  • Organizer: 第67回日本ウイルス学会学術集会
• [Presentation] ロタウイルス人工合成法の開発と新規ワクチン (2018)
  • Author(s): Kobayashi T.
  • Organizer: 第22回日本ワクチン学会学術集会
  • Invited
• [Presentation] Nelson Bay reovirus σC body domain is associated with strain-specific differences in viral replication (2018)
  • Author(s): Kawagishi T, Kanai Y, Sakai Y, Nouda R, Tani H, Shimojima M, Saijo M, Matsuura Y, Kobayashi T.
  • Organizer: 第66回日本ウイルス学会学術集会
• [Presentation] Platform for rotavirus vaccine development using plasmid-based reverse genetics (2018)
  • Author(s): Kanai Y, Kawagishi T, Onishi M, Pannacha P, Nouda R, Nurdin J, Nomura K, Ushijima H, Kobayashi T.
  • Organizer: 第66回日本ウイルス学会学術集会
• [Presentation] Fusogenic bat-borne orthoreovirus p17 protein regulates viral replication in a host-specific manner (2018)
  • Author(s): Nouda R, Kawagishi T, Kanai Y, Shimojima M, Saijo M, Matsuura Y, Kobayashi T.
  • Organizer: 第66回日本ウイルス学会学術集会
• [Presentation] Development of stable reporter rotaviruses (2018)
  • Author(s): Pannacha P, Kanai Y, Onishi M, Kawagishi T, Nouda R, Nurdin J, Matsuura Y, Kobayashi T.
  • Organizer: 第66回日本ウイルス学会学術集会
• [Presentation] Reverse genetics systems for orthoreoviruses and rotaviruses (2018)
  • Author(s): Kobayashi T.
  • Organizer: 13th International dsRNA Symposium 2018
  • Int'l Joint Research / Invited
• [Presentation] Antigenicity of simian and human reassortant rotaviruses generated by reverse genetics (2018)
  • Author(s): Kanai Y, Kawagishi T, Onishi M, Pannacha P, Nouda R, Nurdin J, Nomura K, Ushijima H, Kobayashi T.
  • Organizer: 13th International dsRNA Symposium 2018
  • Int'l Joint Research
• [Presentation] Nelson Bay Orthoreovirus cell attachment protein σC determines strain-specific differences in viral replication and pathogenesis (2018)
  • Author(s): Kawagishi T, Kanai Y, Yusuke Sakai, Nouda R,Tani H, Shimojima M, Saijo M, Matsuura Y, Kobayashi T.
  • Organizer: 13th International dsRNA Symposium 2018
  • Int'l Joint Research
• [Presentation] 次世代組換えロタウイルスワクチン作製の試み (2018)
  • Author(s): 金井 祐太、川岸 崇裕、Pimfhun Pannacha、納田 遼太郎、Nurdin Jeffery、野村 圭一郎、牛島 廣治、小林 剛
  • Organizer: 第161回日本獣医学会学術集会
• [Presentation] Nelson Bay orthoreovirus p17 protein regulates viral replication in a host-specific manner (2018)
  • Author(s): Kobayashi T.
  • Organizer: 第12回日中国際ウイルス学会
  • Int'l Joint Research
• [Book] Pharama Medica January 2021 vol.39 No1 (2021)
  • Author(s): 南 昌平、小林 剛
  • Total Pages: 6
  • Publisher: メディカルレビュー社
  • ISBN: 9784779224621
• [Book] 腎と透析 (2018)
  • Author(s): 小林 剛
  • Total Pages: 3
  • Publisher: 東京医学社
• [Book] 獣医畜産新報 (2018)
  • Author(s): 川岸 崇裕
  • Total Pages: 7
  • Publisher: 文永堂出版