非洲马瘟病毒VP2蛋白的截短表达及免疫原性分析  

Truncated expression and immunogenicity analysis of VP2 protein of African horse sickness virus

作　　者：王轩莹 范亚亚 户鑫兵 徐婧 张鸿歌 田占成[1] 苟惠天[2] 郑玉姝[3] 关贵全[1] 罗建勋[1] 殷宏[1] 独军政[1,4] WANG Xuanying;FAN Yaya;HU Xinbing;XU Jing;ZHANG Hongge;TIAN Zhancheng;COU Huitian;ZHENG Yushu;CUAN Guiquan;LUO Jianxun;YIN Hong;DU Junzheng(State Key Laboratory for Animal Disease Control and Prevention/College of Veterinary Medicine of Lanzhou University/Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Lanzhou 730000,China;College of Veterinary Medicine,Gansu A gricultural University,Lanzhou 730070,China;College of Animal Science and Veterinary Medicine,Henan Institute of Science and Technology,Xinxiang,453003,China;Gansu Province Research Center for Basic Disciplines of Pathogen Biology,Lanzhou 730046,China)

机构地区：[1]中国农业科学院,兰州兽医研究所,兰州大学动物医学与生物安全学院动物疫病防控全国重点实验室,甘肃兰州730000 [2]甘肃农业大学动物医学院,甘肃兰州730070 [3]河南科技学院动物科技学院,河南新乡453003 [4]甘肃省病原生物学基础学科研究中心,甘肃兰州730046

出　　处：《中国兽医科学》2024年第8期1087-1095,共9页Chinese Veterinary Science

基　　金：国家重点研发计划项目(2021YFD1800500);中央级公益性科研院所基本科研业务费项目(1610312021014);甘肃省基础研究创新群体项目(22JR5RA024);甘肃省科技厅项目(22CX8NA011);农业科技创新工程(CAAS-ASTIP-2016-LVRI)。

摘　　要：为评价非洲马瘟病毒(AHSV)VP2截短蛋白的免疫原性,以人工合成的含有AHSV S2的全长基因为模板,分别构建重组表达质粒p RSF-S2-502(编码1~502 aa)和p RSF-S2-1056(编码503~1056 aa),分别将其转入大肠杆菌BL21(DE3)感受态细胞中进行表达。将纯化后的重组蛋白免疫小鼠后,采用间接ELISA测定小鼠血清中特异性抗体水平,用流式细胞术检测小鼠脾T细胞亚群、CD4^(+)和CD8^(+)T细胞表达IFN-γ的水平。结果显示,与PBS对照组相比,各免疫组小鼠血清中特异性抗体水平均显著升高,脾内CD4^(+)T和CD8^(+)T细胞水平均显著升高;免疫组小鼠脾CD4^(+)T和CD8^(+)T细胞表达IFN-γ的水平均升高。结果表明,重组AHSV VP2截短蛋白对小鼠具有较好的免疫原性,为进一步研究VP2蛋白生物学功能及非洲马瘟病毒亚单位疫苗奠定了理论基础。In order to evaluate the immunogenicity of the truncated VP2 protein of African horse sickness virus(AHSV),the recombinant expression plasmids p RSF-S2-502(encoding 1-502 aa)and p RSF-S2-1056(encoding 503-1056 aa)were constructed using a synthetic full-length gene containing AHSV S2 as a template,and were transformed into Escherichia coli BL21(DE3)cells for specific expression.Mice were immunized with purified protein,and specific antibody levels in serum were determined by indirect ELISA.Flow cytometry was used to detect the T cell subsets,CD4^(+)and CD8^(+)T cells express IFN-γlevels.The results showed that,compared with PBS control group,the specific antibody levels in mice serum of all immunized groups were significantly increased,and the levels of CD4^(+)T and CD8^(+)T in spleen cells were significantly increased.The levels of IFN-γexpressed by CD4^(+)and CD8^(+)T cells in spleen cells of immunized mice groups were increased.The result showed that the purified truncated AHSV VP2 protein could produce good immunogenicity in mice,which laid a theoretical foundation for further studying on the biological function of VP2 protein and the development of AHSV subunit vaccine.

关 键 词：非洲马瘟病毒 VP2蛋白 截短表达 免疫原性 

分 类 号：S852.659.1[农业科学—基础兽医学]