Construction of recombinant adenovirus expressing capsid protein of serotype O foot-and-mouth disease virus and analysis of its immunogenicity.

Cancan Wang; Liping Zhang; Xinsheng Liu; Peng Zhou; Li Pan; Yonglu Wang

Return

Construction of recombinant adenovirus expressing capsid protein of serotype O foot-and-mouth disease virus and analysis of its immunogenicity.

Author: Cancan WANG ¹ ; Liping ZHANG ¹ ; Xinsheng LIU ¹ ; Peng ZHOU ¹ ; Li PAN ¹ ; Yonglu WANG ¹
Author Information

1. OIE/National Foot-and-Mouth Disease Reference Laboratory, State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, Gansu, China.
Publication Type:Journal Article
Keywords: FMDV serotype O; P12A and 3B3C; immunogenicity; recombinant adenovirus
MeSH: Adenoviridae/genetics*; Adenoviruses, Human/genetics*; Animals; Antibodies, Viral; Capsid/metabolism*; Capsid Proteins; Foot-and-Mouth Disease/prevention & control*; Foot-and-Mouth Disease Virus/genetics*; HEK293 Cells; Humans; Mice; Recombinant Proteins/genetics*; Serogroup; Swine; Viral Proteins; Viral Vaccines/genetics*
From: Chinese Journal of Biotechnology 2022;38(5):1824-1836
CountryChina
Language:Chinese
Abstract: In order to construct a recombinant replication deficient human type 5 adenovirus (Ad5) expressing a foot-and-mouth disease virus (FMDV) capsid protein, specific primers for P12A and 3B3C genes of FMDV-OZK93 were synthesized. The P12A and 3B3C genes were then amplified and connected by fusion PCR, and a recombinant shuttle plasmid pDC316-mCMV-EGFP-P12A3B3C expressing the FMDV-OZK93 capsid protein precursor P12A and 3B3C protease were obtained by inserting the P12A3B3C gene into the pDC316-mCMV-EGFP plasmid. The recombinant adenovirus rAdv-P12A3B3C-OZK93 was subsequently packaged, characterized and amplified using AdMax^TM adenovirus packaging system, and the expression was verified by infecting human embryonic kidney cell HEK-293. The humoral and cellular immunity levels of well-expressed and purified recombinant adenovirus immunized mice were evaluated. The results showed that rAdv-P12A3B3C-OZK93 could be stably passaged and the maximum virus titer reached 1×10^9.1 TCID₅₀/mL. Western blotting and indirect immunofluorescence showed that rAdv-P12A3B3C-OZK93 expressed the FMDV-specific proteins P12A and VP1 in HEK-293 cells. In addition, the PK cell infection experiment confirmed that rAdv-P12A3B3C-OZK93 could infect porcine cells, which is essential for vaccination in pigs. Comparing with the inactivated vaccine group, the recombinant adenovirus could induce higher FMDV-specific IgG antibodies, γ-IFN and IL-10. This indicates that the recombinant adenovirus has good immunity for animal, which is very important for the subsequent development of foot-and-mouth disease vaccine.