New attempt of cold-chain free oral influenza vaccine development

• Project/Area Number: 24659203
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Virology
• Research Institution: The University of Tokyo
• Principal Investigator: YUKI Yoshikazu 東京大学, 医科学研究所, 助教 (60345030)
• Research Collaborator: FUKUYAMA Yoshiko ; MEJIMA Mio ; KUROKAWA Shiho ; SUZUKI Yuji
• Project Period (FY): 2012-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2014: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2013: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2012: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
• Keywords: ワクチン

Outline of Final Research Achievements

By using a T-DNA vector driven by a prolamin promoter and an overexpression vaccine cassette with RNAi, we have established MucoRice-CTB-HA1 intended as a cold-chain free oral influenza vaccine. To investigate the immunogenicity of MucoRice-CTB-HA1, we subcutaneously or orally immunized mice. Although subcutaneous immunization induced CTB- and HA-specific IgG immune responses, oral immunization induced CTB-specific IgG and mucosal IgA immune responses but not HA-specific one. We now consider to improve the immunization system by using subcutaneous followed by oral immunization to induce systemic and mucosal HA-specific Ab immune responses.

Research Products

• [Journal Article] Determination of genomic location and structure of the transgenes in marker-free rice-based cholera vaccine by using whole genome resequencing approach. (2015)
  • Author(s): Mejima, M., Kashima, K., Kuroda, M., Takeyama, N., Kurokawa S., Fukuyama Y., Kiyono H., Itoh, K., Mitsui, T., and Yuki, Y.
  • Journal Title: Plant Cell Tiss. Organ Cult. Volume: 120 Issue: 1 Pages: 35-48
  • DOI: 10.1007/s11240-014-0575-4
  • Peer Reviewed
• [Journal Article] Nanogel-based pneumococcal surface protein A nasal vaccine induces microRNA-associated Th17 cell responses with neutralizing antibodies against Streptococcus pneumoniae in macaques. (2015)
  • Author(s): Fukuyama, Y., Yuki, Y., Katakai, Y., Harada, N., Takahashi, H., Takeda, S., Mejima, M., Joo, S., Kurokawa, S., Sawada, S., Shibata, H., Park, EJ., Fujihashi., K, Briles, D., Yasutomi, Y., Tsukada, H., Akiyoshi, K., Kiyono, H.
  • Journal Title: Mucosal Immunol. Volume: - Issue: 5 Pages: 1144-1153
  • DOI: 10.1038/mi.2015.5
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Comparative whole-genome analyses of selection marker‐free rice-based cholera toxin B-subunit vaccine lines and wild-type lines. (2015)
  • Author(s): Kashima, K., Mejima, M., Kurokawa, S., Kuroda, M., Kiyono, H. and Yuki, Y.
  • Journal Title: BMC Genomics Volume: 48 Issue: 1 Pages: 48-48
  • DOI: 10.1186/s12864-015-1285-y
  • Peer Reviewed / Open Access
• [Journal Article] Novel transgenic rice-based vaccines. (2015)
  • Author(s): Azegami, T., Itoh, H., Kiyono, H. and Yuki, Y.
  • Journal Title: Arch. Immunol. Ther. Exp. Volume: 63(2) Issue: 2 Pages: 87-99
  • DOI: 10.1007/s00005-014-0303-0
  • Peer Reviewed / Open Access
• [Journal Article] Specific expression of apolipoprotein A-IV in the follicle-associated epithelium of the small intestine. (2014)
  • Author(s): Tokuhara, D., Nochi, T., Matsumura, A., Mejima, M., Takahashi, Y., Kurokawa, S., Kiyono, H. and Yuki Y.
  • Journal Title: Dig. Dis. Sci. Volume: 59 Issue: 11 Pages: 2682-2692
  • DOI: 10.1007/s10620-014-3203-6
  • Peer Reviewed / Open Access
• [Journal Article] RNAi-mediated suppression of endogenous storage proteins leadsto a change in localization of overexpressed cholera toxinB-subunit and the allergen protein RAG2 in rice seeds. (2014)
  • Author(s): Kurokawa, S., et al.
  • Journal Title: Plant Cell Rep. Volume: 33 Issue: 1 Pages: 75-87
  • DOI: 10.1007/s00299-013-1513-3
  • Peer Reviewed
• [Journal Article] A rice-based soluble form of a murine TNF-specific ll ama variable domain of heavy-chain antibody suppresses collagen-induced arthr itis in mice. (2014)
  • Author(s): M. Abe, Y. Yuki*, S. Kurokawa et.al.
  • Journal Title: J. Biotechnol. Volume: 175 Pages: 45-52
  • Peer Reviewed
• [Journal Article] Induction of toxin-specific neutralizing immunity by molecularly uniform rice-based oral cholera toxin B subunit vaccine without plant-associated sugar modification. (2013)
  • Author(s): Yuki, Y., et al.
  • Journal Title: Plant Biotechnol. J. Volume: 11 Issue: 7 Pages: 799-808
  • DOI: 10.1111/pbi.12071
  • Peer Reviewed
• [Journal Article] MucoRice-cholera toxin B-subunit, a rice-based oral cholera vaccine, down-regulates the expression of α-amylase/trypsin inhibitor-like protein family as major rice allergens. (2013)
  • Author(s): Kurokawa, S., et al.
  • Journal Title: J. Proteome Res. Volume: 12 Issue: 7 Pages: 3372-3382
  • DOI: 10.1021/pr4002146
  • Peer Reviewed
• [Journal Article] Rice-based oral antibody fragment prophylaxis and therapy against rotavirus infection. (2013)
  • Author(s): Tokuhara, D., et al.
  • Journal Title: J. Clin. Invest. Volume: 123 Issue: 9 Pages: 3829-3838
  • DOI: 10.1172/jci70266
  • Peer Reviewed
• [Journal Article] Nanogel-Based PspA Intranasal Vaccine Prevents Invasive Disease and Nasal colnization by Streptococcus pneumoniae (2013)
  • Author(s): IG. Kong, A. Sato, Y. Yuki, et.al.
  • Journal Title: Infection and Immunity Volume: 81 Pages: 1825-1634
  • Peer Reviewed
• [Journal Article] Transcription factor Spi-B–dependent and –independentpathways for the development of Peyer’s patch M cells. (2012)
  • Author(s): S. Sato, S. Kaneto, N. Shibata, Y. Takahashi, H. Okura, J. Kunisawa,, Y. Yuki and H.Kiyono
  • Journal Title: Mucosal Immunol Volume: Dec 5
  • Peer Reviewed
• [Journal Article] RNAi suppression of rice endogenous storage proteins enhances the production of rice-based Botulinum neutrotoxin type A vaccine (2012)
  • Author(s): Y. Yuki, M. Mejima, S. Kurokawa, et.al.
  • Journal Title: Vaccine Volume: 30 Pages: 4160-4166
  • Peer Reviewed
• [Journal Article] Novel vaccine development strategies for inducing mucosal immunity
  • Author(s): Fujkuyama Y, Tokuhara D, Kataoka K, Gilbert RS, McGhee JR, Yuki Y, et al.
  • Journal Title: Expert Rev. Vaccines Volume: 11 Pages: 367-379
  • Peer Reviewed
• [Presentation] Critical role of TSLP-TSLPR interaction in inducing secretory IgA responses after mucosal immunization (2014)
  • Author(s): J. Sunyi, Y. Fukuyama, Y. Yuki, et.al.
  • Organizer: 日本免疫学会
  • Place of Presentation: 京都
  • Year and Date: 2014-12-10 – 2014-12-12
• [Presentation] アジュバントフリーナノゲル型肺炎球菌経鼻ワクチンのサルでの免疫効果と安全性 (2014)
  • Author(s): 幸 義和、清野宏
  • Organizer: 日本ワクチン学会
  • Place of Presentation: 福岡
  • Year and Date: 2014-12-06 – 2014-12-07
• [Presentation] Establishment of selection marker-free rice-based oral cholera toxin B-subunit vaccine and characterization of location and structure of transgene by using whole genome resequencing analysis (2014)
  • Author(s): Y.Yuki, M. Mejima, K.Kashima et.al.
  • Organizer: International Association for Plant Biotechnology Congress
  • Place of Presentation: Melbourne, Australia
  • Year and Date: 2014-08-11 – 2014-08-15
• [Presentation] Whole genome analysis of selection marker-free MucoRice-CTB, a rice-based oralcholera vaccine (2014)
  • Author(s): K.Kashima, M. Mejima, &Y. Yuk
  • Organizer: International Association for Plant Biotechnology Congress
  • Place of Presentation: Melbourne, Australia
  • Year and Date: 2014-08-11 – 2014-08-15
• [Presentation] Molecularly Uniform Rice-based Oral Cholera Toxin B Subunit Vaccine without Plant-associated Sugar Modification induces toxin-specific neutralizing immunity in mice and macaques
  • Author(s): Y. Yuki, M. Mejima, S. Kurokawa et.al.
  • Organizer: 15th International congress of immunology
  • Place of Presentation: Milan, Italy
• [Presentation] Nanogel-based PspA nasal vaccine induces S. pneumoniae-specific neutralizing antibody immune responses in non-human primates.
  • Author(s): Y. Fukuyama, Y. Yuki, Y. Katakai et.al
  • Organizer: 15th International congress of immunology
  • Place of Presentation: Milan, Italy
• [Presentation] Establishment of murine norvirus S7 infection system for vaccine development
  • Author(s): N. Takayama, Y. Chen et.al.. & Y. Yuki
  • Organizer: 15th International congress of immunology
  • Place of Presentation: Milan, Italy
• [Presentation] 幸 義和、清野 宏：経鼻投与された粘膜アジュバントであるコレラトキシンは嗅覚神経を破壊する
  • Author(s): 幸 義和、清野 宏
  • Organizer: 日本ワクチン学会
  • Place of Presentation: 津、三重県
• [Presentation] Regional T memory and miRNA biomarkers revealed by oral vaccination with MucoRice-CTB
  • Author(s): EJ Park, Y. Yuki, H, Kiyono
  • Organizer: 日本免疫学会
  • Place of Presentation: 幕張、千葉県
• [Presentation] Adjuvant-free nanogel-based PspA nasal vaccine for the induction of protective immunity against Pneumococcus
  • Author(s): Y. Yuki, IG. Kong, A. Sato,et.al.
  • Organizer: The American Association of Immunologists
  • Place of Presentation: Boston, USA
• [Presentation] ブタ大腸菌性下痢症予防食べるワクチンの開発研究
  • Author(s): 幸 義和
  • Organizer: 動物ワクチンーバイオ医薬研究会
  • Place of Presentation: 盛岡
  • Invited
• [Presentation] MucoRice-CTB as an oral vaccine for the prevention of entroxigenic E. coli-mediated diarrhea in pigs.
  • Author(s): N. Takeyama, K. Oroku, D. Tokuhara, S. Nagai, H. Kiyono, Y. Yuki.
  • Organizer: 日本ワクチン学会
  • Place of Presentation: 東京
• [Presentation] Heat tolerance evaluation of the rice type oral cholera vaccine, MucoRice-CTB, for a clinical study
  • Author(s): K. Kashima, H. Hiroiwa, Y. Yuki, H. Kiyono.
  • Organizer: 日本ワクチン学会
  • Place of Presentation: 東京
• [Presentation] Characterization of effector/memory CD4 T cells expanded in the mice vaccinated with MucoRice-CTB
  • Author(s): EJ. Park, S. Joo, S. Kurokawa, H. Kiyono, Y. Yuki.
  • Organizer: 日本ワクチン学会
  • Place of Presentation: 東京
• [Book] 注射剤・経口製剤に代る新しい薬剤投与ヂバイスの開発― (2014)
  • Author(s): 幸 義和
  • Publisher: 技術情報協会
• [Book] 粘膜ワクチン製剤のＤＤＳ技術の動向と実用化の可能性― (2013)
  • Author(s): 幸 義和
  • Publisher: 技術情報協会
• [Book] ワクチンのDDS技術の開発動向―経鼻投与技術動向― 「ワクチンの市場動向と開発・製造実務集」 (2012)
  • Author(s): 幸 義和ら 多数
  • Publisher: 技術情報協会
• [Patent(Industrial Property Rights)] 肺炎球菌経鼻ワクチン (2014)
  • Inventor(s): 幸義和、清野宏、澤田晋一、秋吉一成
  • Industrial Property Rights Holder: 東京大学ＴＬＯ
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2014-027205
  • Filing Date: 2014-02-17