Chen Liyi, Zhang Xinheng, Shao Guanming, Shao Yangyang, Hu Zezhong, Feng Keyu, Xie Zi, Li Hongxin, Chen Weiguo, Lin Wencheng, Yuan Hengxing, Wang Hailong, Fu Jun, Xie Qingmei
Heyuan Branch, Guangdong Provincial Laboratory of Lingnan Modern Agricultural Science and Technology and Guangdong Provincial Key Lab of Agro-Animal Genomics and Molecular Breeding, College of Animal Science, South China Agricultural University, Guangzhou, China.
Guangdong Engineering Research Center for Vector Vaccine of Animal Virus, Guangzhou, China.
Front Vet Sci. 2022 Apr 13;9:832255. doi: 10.3389/fvets.2022.832255. eCollection 2022.
African swine fever (ASF) is a highly contact infectious disease caused by the African swine fever virus (ASFV). The extremely complex structure and infection mechanism make it difficult to control the spread of ASFV and develop the vaccine. The ASFV genome is huge with many antigenic genes. Among them, (p30), (pp62), (p54), (p72), and (CD2v) are involved in the process of the virus cycle, with strong immunogenicity and the ability to induce the body to produce neutralizing antibodies. In this study, the recombinant virus rBartha-K61-pASFV that expresses the above ASFV antigen genes was constructed by Red/ET recombineering technology using pseudorabies virus (PRV) vaccine strain Bartha-K61. Western blot analysis showed that the ASFV antigen gene was expressed and the recombinant virus showed good genetic stability and proliferation characteristics in 15 continuous generations on porcine kidney (PK15) cells. The results of immunoassay of piglets and mice showed that rBartha-K61-pASFV had good immunogenicity and could induce higher antibody levels in the body. Therefore, PRV was a promising viral vector for expressing the ASFV antigen gene, and all the experiments in this study laid a foundation for the further development of a new viral vector vaccine of ASFV.
非洲猪瘟（ASF）是一种由非洲猪瘟病毒（ASFV）引起的高度接触性传染病。其极其复杂的结构和感染机制使得控制ASFV的传播以及研发疫苗变得困难。ASFV基因组庞大，含有许多抗原基因。其中，p30、pp62、p54、p72和CD2v参与病毒循环过程，具有很强的免疫原性以及诱导机体产生中和抗体的能力。在本研究中，利用伪狂犬病病毒（PRV）疫苗株Bartha-K61，通过Red/ET重组技术构建了表达上述ASFV抗原基因的重组病毒rBartha-K61-pASFV。蛋白质免疫印迹分析表明，ASFV抗原基因得到表达，并且该重组病毒在猪肾（PK15）细胞上连续传代15代时表现出良好的遗传稳定性和增殖特性。仔猪和小鼠的免疫测定结果表明，rBartha-K61-pASFV具有良好的免疫原性，能够在体内诱导产生更高的抗体水平。因此，PRV是一种有前景的用于表达ASFV抗原基因的病毒载体，本研究中的所有实验为进一步研发新型ASFV病毒载体疫苗奠定了基础。
