Development of a Japanese encephalitis virus-like particle vaccine in silkworms using codon-optimised prM and envelope genes
Sayaka Matsuda,
Reiko Nerome,
Kenichi Maegawa,
Akira Kotaki,
Shigeo Sugita,
Kazunori Kawasaki,
Kazumichi Kuroda,
Ryoji Yamaguchi,
Tomohiko Takasaki,
Kuniaki Nerome
Affiliations
Sayaka Matsuda
The Institute of Biological Resources, 893-2, Nakayama, Nago-shi, Okinawa 905-0004, Japan
Reiko Nerome
The Institute of Biological Resources, 893-2, Nakayama, Nago-shi, Okinawa 905-0004, Japan
Kenichi Maegawa
The Institute of Biological Resources, 893-2, Nakayama, Nago-shi, Okinawa 905-0004, Japan
Akira Kotaki
The Institute of Biological Resources, 893-2, Nakayama, Nago-shi, Okinawa 905-0004, Japan
Shigeo Sugita
Equine Research Institute, Japan Racing Association, 1400-4, Shiba, Shimotsuke-shi, Tochigi 329-0412, Japan
Kazunori Kawasaki
National Institute of Advanced Science and Technology (AIST), 1-8-31, Midorigaoka, Ikeda, Osaka 563-8577, Japan
Kazumichi Kuroda
Division of Microbiology, Nihon University School of Medicine, 30-1, Oyaguchi-kamicho, Itabashi-ku, Tokyo 173-8610, Japan
Ryoji Yamaguchi
Laboratory of Veterinary Pathology, Department of Veterinary, Faculty of Agriculture, University of Miyazaki, 1-1 Gakuenkibanadai-Nishi, Miyazaki 889-2192, Japan
Tomohiko Takasaki
Department of Virology 1, National Institute of Infectious Diseases, 1-23-1 Toyama, Shinjuku-ku, Tokyo 162-8640, Japan
Kuniaki Nerome
The Institute of Biological Resources, 893-2, Nakayama, Nago-shi, Okinawa 905-0004, Japan; Corresponding author.
We have successfully prepared a Japanese encephalitis virus (JEV) − Nakayama virus like particle (NVLP) vaccine using synthetic codon-optimized prM and E genes. The expression of the recombinant JEV Nakayama-BmNPV (JEV-NNPV) virus was determined in infected silkworm Bm-N cells by fluorescence and Western blot analysis. The recombinant was inoculated into silkworm pupae and the yield of Nakayama VLP (NVLP) reached a peak in the homogenates after 3 days. Additionally, in the peptide analysis of infected pupae homogenate, it appeared approximately 300–500 μg E protein/pupa were produced. When purified the above eluates on the discontinuous sucrose density gradient centrifugation, NVLP showed a strong hemagglutination (HA) activity by using chicken red blood cell in phosphate-buffered saline (PBS) free from Mg++ and Ca++ ions. The immune antisera against NVLP strain could efficiently neutralize the plaque formation of Nakayama, Beijing-1 and Muar strains, showing tendency of much higher reaction with heterologous Muar strain than homologous Nakayama strain. Our findings suggest that the JEV-NVLP may be useful for JEV epidemic control in many endemic areas of Asian countries as a widely effective and less expensive JE vaccine.
