Preparation and Immunogenicity Studies on mRNA Vaccine Based on VP1 Gene of Porcine foot-and-mouth disease type O virus
DONG Jin-Jie1,2,3,4, WANG Hui-Bao2, WANG Fan2, DENG Rui-Xue2, LIU Ping2, CHEN Miao-Miao2, LI Yang-Fan2, ZHANG Yong1,4,*
1 College of Veterinary Medicine, Gansu Agricultural University, Lanzhou 730070, China; 2 China Agricultural Vet. Bio. Science and Technology Co., Ltd., Lanzhou 730046, China; 3 Lanzhou Veterinary Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China; 4 Gansu Key Laboratory of Animal Generational Physiology and Reproductive Regulation
Abstract：Foot-and-mouth disease (FMD) is one of the most important infectious diseases causing significant economic losses to the swine (Sus scrofa domesticus) industry. Vaccine immunization is still the one of main ways to prevent and control foot-and-mouth disease. Because Foot-and-mouth disease virus is easy to mutate, the antigen mismatch between candidate vaccine strains and epidemic strains will affect the immune efficacy of vaccine and the effect of disease control. According to the molecular and functional characteristics of structural protein VP1 (viral structural protein) gene, using the inactivated Porcine foot-and-mouth disease virus type O strain (O /MYA98/BY/2010) as the template, the structural protein VP1 gene was amplified and ligated into pSFV1 vector to construct the recombinant plasmid pSFV1-VP1. To obtain mRNA, the recombinant plasmid was treated by linearization and in vitro transcription, then the mRNA was transfected into baby hamster syrian kidney-21 (BHK-21) cells and verified its expression properly. After that, it was mixed with the liposome nanoparticles to immunize guinea pigs (Cavia porcellus), and the immune response and efficacy were detected. The results showed that the mRNA was expressed in BHK-21 cells after 24 h post-transfection evaluated by immunofluorescence assay (IFA) and Western blot; In guinea pigs immunized with the mRNA vaccine, specific antibody, neutralizing antibody, interleukin 4 (IL-4) and γ interferon (IFN-γ) in serum were significantly higher than those of guinea pigs. The mRNA vaccine could provide 4/4 protection against 50 times GPID50 (50% Guinea pig infectious dose) virus challenge in vaccinated guinea pigs, implying humoral and cellular immune response elicited by the developed mRNA vaccine with sound immunogenicity, which provides a new route for the development of foot-and-mouth disease emergency vaccine.
董金杰, 王会宝, 王凡, 邓瑞雪, 刘萍, 陈苗苗, 李扬帆, 张勇. 基于猪口蹄疫O型病毒结构蛋白VP1基因的mRNA疫苗制备及免疫活性研究[J]. 农业生物技术学报, 2022, 30(6): 1219-1227.
DONG Jin-Jie, WANG Hui-Bao, WANG Fan, DENG Rui-Xue, LIU Ping, CHEN Miao-Miao, LI Yang-Fan, ZHANG Yong. Preparation and Immunogenicity Studies on mRNA Vaccine Based on VP1 Gene of Porcine foot-and-mouth disease type O virus. 农业生物技术学报, 2022, 30(6): 1219-1227.
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