State Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, China.
Viruses. 2023 Jun 28;15(7):1467. doi: 10.3390/v15071467.
A cell line expressing the CD2v protein of ASFV was generated. The efficient expression of CD2v protein was determined by immunofluorescence and Western blotting. The CD2v protein was Ni-affinity purified from the supernatant of cell cultures. The CD2v-expressing cells showed properties of hemadsorption, and the secreted CD2v protein exhibited hemagglutinating activity. The antigenicity and immunoprotection ability of CD2v were evaluated by immunizing pigs alone, combined with a cell-line-expressed p30 protein or triple combined with p30 and K205R protein. Immunized pigs were challenged with the highly virulent ASFV strain HLJ/18. Virus challenge results showed that CD2v immunization alone could provide partial protection at the early infection stage. Protein p30 did not show synergistic protection effects in immunization combined with CD2v. Interestingly, immunization with the triple combination of CD2V, p30 and K205R reversed the protection effect. The viremia onset time was delayed, and one pig out of three recovered after the challenge. The pig recovered from ASFV clinical symptoms, the rectal temperature returned to normal levels and the viremia was cleared. The mechanism of this protection effect warrants further investigation.
表达 ASFv CD2v 蛋白的细胞系已被构建。通过免疫荧光和 Western blot 确定了 CD2v 蛋白的有效表达。从细胞培养物的上清液中用 Ni 亲和纯化 CD2v 蛋白。表达 CD2v 的细胞表现出红细胞吸附特性，分泌的 CD2v 蛋白具有血凝活性。通过单独免疫猪、与细胞系表达的 p30 蛋白联合免疫或与 p30 和 K205R 蛋白联合免疫三种方式评估 CD2v 的抗原性和免疫保护能力。免疫猪用高致病性 ASFv 株 HLJ/18 进行攻毒。病毒攻毒结果表明，单独免疫 CD2v 可在早期感染阶段提供部分保护。在与 CD2v 联合免疫时，蛋白 p30 没有表现出协同保护作用。有趣的是，CD2V、p30 和 K205R 的三联免疫逆转了保护作用。病毒血症发病时间延迟，3 头猪中有 1 头在攻毒后恢复。从 ASF 临床症状中恢复的猪，直肠温度恢复正常水平，病毒血症清除。这种保护作用的机制需要进一步研究。
