Stable trimer formation of spike protein from porcine epidemic diarrhea virus improves the efficiency of secretory production in silkworms and induces neutralizing antibodies in mice

Veterinary Research

Takeshi Miyata¹  Jae Man Lee²  Takahiro Kusakabe³  Akitsu Masuda³  Hiroaki Mon³  Takeru Ebihara³  Kohei Kakino³  Masato Hino⁴  Ryosuke Fujita⁴ 

[1] Department of Biochemistry and Biotechnology, Faculty of Agriculture, Kagoshima University, 1-21-24 Korimoto, 890-0065, Kagoshima, Japan;Laboratory of Creative Science for Insect Industries, Kyushu University Graduate School of Bioresource and Bioenvironmental Sciences, Motooka 744, Nishi-ku, 819-0395, Fukuoka, Japan;Laboratory of Insect Genome Science, Kyushu University Graduate School of Bioresource and Bioenvironmental Sciences, Motooka 744, Nishi-ku, 819-0395, Fukuoka, Japan;Laboratory of Sanitary Entomology, Kyushu University Graduate School of Bioresource and Bioenvironmental Sciences, Motooka 744, Nishi-ku, 819-0395, Fukuoka, Japan;

关键词: Porcine epidemic diarrhea virus;    spike protein;    trimeric motif;    silkworm-baculovirus expression vector system;    subunit vaccine;   

DOI  :  10.1186/s13567-021-00971-5

来源: Springer

【 摘 要 】

Porcine epidemic diarrhea virus (PEDV) is a highly infectious pathogen of watery diarrhea that causes serious economic loss to the swine industry worldwide. Especially because of the high mortality rate in neonatal piglets, a vaccine with less production cost and high protective effect against PEDV is desired. The intrinsically assembled homotrimer of spike (S) protein on the PEDV viral membrane contributing to the host cell entry is a target of vaccine development. In this study, we designed trimerized PEDV S protein for efficient production in the silkworm-baculovirus expression vector system (silkworm-BEVS) and evaluated its immunogenicity in the mouse. The genetic fusion of the trimeric motif improved the expression of S protein in silkworm-BEVS. A small-scale screening of silkworm strains to further improve the S protein productivity finally achieved the yield of about 2 mg from the 10 mL larval serum. Mouse immunization study demonstrated that the trimerized S protein could elicit strong humoral immunity, including the S protein-specific IgG in the serum. These sera contained neutralizing antibodies that can protect Vero cells from PEDV infection. These results demonstrated that silkworm-BEVS provides a platform for the production of trimeric S proteins, which are promising subunit vaccines against coronaviruses such as PEDV.

【 授权许可】

CC BY