Frontiers in Plant Science (Mar 2024)
MucoRice-CTB line 19A, a new marker-free transgenic rice-based cholera vaccine produced in an LED-based hydroponic system
- Yoshikazu Yuki,
- Yoshikazu Yuki,
- Yoshikazu Yuki,
- Shiho Kurokawa,
- Shiho Kurokawa,
- Kotomi Sugiura,
- Kotomi Sugiura,
- Koji Kashima,
- Shinichi Maruyama,
- Tomoyuki Yamanoue,
- Tomoyuki Yamanoue,
- Ayaka Honma,
- Mio Mejima,
- Natsumi Takeyama,
- Natsumi Takeyama,
- Masaharu Kuroda,
- Hiroko Kozuka-Hata,
- Masaaki Oyama,
- Takehiro Masumura,
- Rika Nakahashi-Ouchida,
- Rika Nakahashi-Ouchida,
- Rika Nakahashi-Ouchida,
- Kohtaro Fujihashi,
- Kohtaro Fujihashi,
- Kohtaro Fujihashi,
- Kohtaro Fujihashi,
- Takashi Hiraizumi,
- Eiji Goto,
- Hiroshi Kiyono,
- Hiroshi Kiyono,
- Hiroshi Kiyono,
- Hiroshi Kiyono,
- Hiroshi Kiyono,
- Hiroshi Kiyono
Affiliations
- Yoshikazu Yuki
- Division of Mucosal Immunology, IMSUT Distinguished Professor Unit, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan
- Yoshikazu Yuki
- R&D department, HanaVax Inc., Chiba, Japan
- Yoshikazu Yuki
- Department of Human Mucosal Vaccinology, Chiba University Hospital, Chiba, Japan
- Shiho Kurokawa
- Division of Mucosal Immunology, IMSUT Distinguished Professor Unit, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan
- Shiho Kurokawa
- Department of Human Mucosal Vaccinology, Chiba University Hospital, Chiba, Japan
- Kotomi Sugiura
- Division of Mucosal Immunology, IMSUT Distinguished Professor Unit, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan
- Kotomi Sugiura
- Department of Human Mucosal Vaccinology, Chiba University Hospital, Chiba, Japan
- Koji Kashima
- Technical Research Institute, Asahi Kogyosha Co., Ltd., Tokyo, Japan
- Shinichi Maruyama
- Technical Research Institute, Asahi Kogyosha Co., Ltd., Tokyo, Japan
- Tomoyuki Yamanoue
- Division of Mucosal Immunology, IMSUT Distinguished Professor Unit, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan
- Tomoyuki Yamanoue
- Department of Human Mucosal Vaccinology, Chiba University Hospital, Chiba, Japan
- Ayaka Honma
- Division of Mucosal Immunology, IMSUT Distinguished Professor Unit, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan
- Mio Mejima
- Division of Mucosal Immunology, IMSUT Distinguished Professor Unit, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan
- Natsumi Takeyama
- Division of Mucosal Immunology, IMSUT Distinguished Professor Unit, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan
- Natsumi Takeyama
- Research Department, Nisseiken Co., Ltd., Tokyo, Japan
- Masaharu Kuroda
- Division of Genome Editing Research, National Agriculture and Food Research Organization, Tsukuba, Japan
- Hiroko Kozuka-Hata
- Medical Proteomics Laboratory, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan
- Masaaki Oyama
- Medical Proteomics Laboratory, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan
- Takehiro Masumura
- Laboratory of Genetic Engineering, Graduate School of Agriculture, Kyoto Prefectural University, Kyoto, Japan
- Rika Nakahashi-Ouchida
- Division of Mucosal Immunology, IMSUT Distinguished Professor Unit, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan
- Rika Nakahashi-Ouchida
- Department of Human Mucosal Vaccinology, Chiba University Hospital, Chiba, Japan
- Rika Nakahashi-Ouchida
- Future Mucosal Vaccine Research and Development Synergy Institute, Chiba University, Chiba, Japan
- Kohtaro Fujihashi
- Division of Mucosal Immunology, IMSUT Distinguished Professor Unit, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan
- Kohtaro Fujihashi
- Department of Human Mucosal Vaccinology, Chiba University Hospital, Chiba, Japan
- Kohtaro Fujihashi
- Future Mucosal Vaccine Research and Development Synergy Institute, Chiba University, Chiba, Japan
- Kohtaro Fujihashi
- 0Department of Pediatric Dentistry, The University of Alabama at Birmingham, Birmingham, AL, United States
- Takashi Hiraizumi
- Technical Research Institute, Asahi Kogyosha Co., Ltd., Tokyo, Japan
- Eiji Goto
- 1Graduate School of Horticulture, Chiba University, Chiba, Japan
- Hiroshi Kiyono
- Division of Mucosal Immunology, IMSUT Distinguished Professor Unit, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan
- Hiroshi Kiyono
- R&D department, HanaVax Inc., Chiba, Japan
- Hiroshi Kiyono
- Department of Human Mucosal Vaccinology, Chiba University Hospital, Chiba, Japan
- Hiroshi Kiyono
- Medical Proteomics Laboratory, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan
- Hiroshi Kiyono
- 2Mucosal Immunology and Allergy Therapeutics, Institute for Global Prominent Research, Research Institute of Disaster Medicine, Chiba University Future Medicine Education and Research Organization, Chiba University, Chiba, Japan
- Hiroshi Kiyono
- 3CU-UCSD Center for Mucosal Immunology, Allergy, and Vaccine (cMAV), Division of Gastroenterology, Department of Medicine, University of California, San Diego, San Diego, CA, United States
- DOI
- https://doi.org/10.3389/fpls.2024.1342662
- Journal volume & issue
-
Vol. 15
Abstract
We previously established the selection-marker-free rice-based oral cholera vaccine (MucoRice-CTB) line 51A for human use by Agrobacterium-mediated co-transformation and conducted a double-blind, randomized, placebo-controlled phase I trial in Japan and the United States. Although MucoRice-CTB 51A was acceptably safe and well tolerated by healthy Japanese and U.S. subjects and induced CTB-specific antibodies neutralizing cholera toxin secreted by Vibrio cholerae, we were limited to a 6-g cohort in the U.S. trial because of insufficient production of MucoRice-CTB. Since MucoRice-CTB 51A did not grow in sunlight, we re-examined the previously established marker-free lines and selected MucoRice-CTB line 19A. Southern blot analysis of line 19A showed a single copy of the CTB gene. We resequenced the whole genome and detected the transgene in an intergenic region in chromosome 1. After establishing a master seed bank of MucoRice-CTB line 19A, we established a hydroponic production facility with LED lighting to reduce electricity consumption and to increase production capacity for clinical trials. Shotgun MS/MS proteomics analysis of MucoRice-CTB 19A showed low levels of α-amylase/trypsin inhibitor-like proteins (major rice allergens), which was consistent with the data for line 51A. We also demonstrated that MucoRice-CTB 19A had high oral immunogenicity and induced protective immunity against cholera toxin challenge in mice. These results indicate that MucoRice-CTB 19A is a suitable oral cholera vaccine candidate for Phase I and II clinical trials in humans, including a V. cholerae challenge study.
Keywords
