伪狂犬病病毒CD8^(+)T细胞表位的鉴定  

Characterization of Pseudorabies Virus CD8^(+)T Cell Epitopes

作　　者：黄香梅 王旭英 耿鑫梅 奚媖牡 廖奕洁 张广欣 莫筱可 陈樱[1,2,3] 欧阳康 韦祖樟[1,2,3] 黄伟坚 覃一峰[1,2,3] HUANG Xiang-mei;WANG Xu-ying;GENG Xin-mei;XI Ying-mu;LIAO Yi-jie;ZHANG Guang-xin;MO Xiao-ke;CHEN Ying;OUYANG Kang;WEI Zu-zhang;HUANG Wei-jian;QIN Yi-feng(Laboratory of Animal infectious Diseases and molecular Immunology,College of Animal Science and Technology,Guangxi University,Nanning,Guangxi 530005,China;Guangxi Zhuang Autonomous Region Engineering Research Center of Veterinary Biologics,Nanning,Guangxi 530005,China;Guangxi Key Laboratory of Animal Breeding,Disease Control and Prevention,Nanning,Guangxi 530004,China;College of Animal Veterinary Medicine,Hebei Agricultural University,Baoding,Hebei 071001,China)

机构地区：[1]广西大学动物科学技术学院,动物传染病与分子免疫学实验室,广西南宁530005 [2]广西壮族自治区兽用生物制品工程研究中心,广西南宁530005 [3]广西畜禽繁育与疾病防控重点实验室,广西南宁530004 [4]河北农业大学动物医学院,河北保定071001

出　　处：《动物医学进展》2025年第4期52-58,共7页Progress In Veterinary Medicine

基　　金：广西研究生教育创新计划资助项目(YCSW2023080)。

摘　　要：伪狂犬病病毒(PRV)是危害养猪业的重要病原,但目前对其感染后诱导的细胞免疫尤其是T细胞抗原表位的鉴定方面研究较少。该研究以C57BL/6小鼠作为PRV感染的动物模型,利用生物信息学软件预测并合成了16条PRV gB、gC和gD蛋白上的H-2b限制性CD8^(+)T细胞表位,经ELISpot、胞内细胞因子染色和CFSE细胞增殖试验检测发现,位于gD蛋白上的多肽^(371)CVYIFFRL^(378)可显著激活CD8^(+)T细胞产生IFN-γ,并促进CD8^(+)T细胞明显增殖,说明该表位属于PRV特异性CD8^(+)T细胞表位。将该表位氨基酸序列与NCBI上16株参考毒株及作者实验室2株PRV毒株的gD蛋白氨基酸序列进行序列比对,发现研究鉴定的T细胞表位高度保守,提示该表位可刺激机体产生对不同亚型PRV感染的交叉保护力。研究结果将为PRV表位疫苗的研制及PRV感染与免疫机制研究奠定基础。Pseudorabies virus(PRV)is an important pathogen jeopardizing the swine industry,but few studies have been conducted to characterize the cellular immunity,especially T cell antigenic epitopes,induced by its infection.In this study,C57BL/6 mice were used as a small animal model of PRV infection,and 16 H-2b-restricted CD8^(+)T-cell epitopes on PRV gB,gC,and gD proteins were predicted and synthesized using bioinformatics software,and the peptides located on the gD proteins were detected by ELISpot,intracellular cytokine staining,and CFSE cell proliferation assay^(371)CVYIFFRL^(378)could significantly activate CD8^(+)T cells to produce IFN-γand promote CD8^(+)T cells to undergo significant proliferation,indicating that this epitope belongs to the PRV-specific CD8^(+)T cell epitope.Sequence comparison of the amino acid sequence of this epitope with the amino acid sequences of the gD protein of the 16 reference strains on NCBI and the two PRV strains in our laboratory revealed that the T-cell epitope identified in this study is highly conserved,suggesting that this epitope may have cross-protective power against different subtypes of PRV.This study will lay the foundation for the development of PRV epitope vaccine and the study of PRV infection and immunity mechanism.

关 键 词：伪狂犬病病毒 CD8^(+)T细胞表位 表位疫苗 

分 类 号：S852.651[农业科学—基础兽医学] S858.28[农业科学—兽医学]