摘要
利用免疫表位数据库(IEDB)预测和筛选非洲猪瘟病毒(ASFV)CP312R蛋白的T细胞表位;通过流式细胞术检测表位肽促进ASFV抗原特异性脾淋巴细胞增殖情况,促进免疫细胞分泌IFN-γ水平及其对T淋巴细胞杀伤作用的影响;通过RT-qPCR技术分析表位肽对ASFV在骨髓巨噬细胞中复制的影响。结果显示,筛选出2条表位肽PP8和PP9,均可促进ASFV抗原特异性脾淋巴细胞增殖(P<0.05),促进CD8^(+)T细胞分泌IFN-γ的水平(P<0.05)。表位肽PP8不能显著促进CD8^(+)T细胞对ASFV感染的靶细胞的杀伤效率(P>0.05),而表位肽PP9能够显著促进CD8^(+)T细胞对靶细胞的杀伤效率(P<0.01)。2条表位肽均可显著抑制ASFV在骨髓巨噬细胞中的复制(P<0.01)。本研究结果为筛选ASFV保护抗原或表位提供了基础数据。
Methods:The T cell epitopes in African swine fever virus CP312R was predicted and screened by immunoepitope database(IEDB).Flow cytometry was carried out to detect the effect of epitope peptide on the proliferation of ASFV antigen-specific spleen lymphocytes,the secretion of IFN-γby different subtypes of immune cells,and T lymphocyte killing.The effect of epitope peptide on ASFV replication in bone marrow-derived macrophages was analyzed by RT-q PCR.The results showed that the epitope peptides PP8 and PP9 promoted the proliferation of ASFV antigen-specific splenic lymphocytes(P<0.05),and increased IFN-γsecretion by CD8^(+)T cells(P<0.05).Epitope peptide PP8 did not significantly promote the killing efficiency of CD8^(+)T cells against ASFV infected target cells(P>0.05)and epitope peptide PP9 can significantly promote the killing efficiency of CD8^(+)T cells to target cells(P<0.01).Both the two epitope peptides inhibited the replication of ASFV in bone marrow-derived macrophages(P<0.01).These results provided basic data for screening protective antigens or epitopes of ASFV.
作者
周晓丽
毛箬青
朱紫祥
孙德惠
朱昱茜
王凌云
齐萌
郑海学
ZHOU Xiao-li;MAO Ruo-qing;ZHU Zi-xiang;SUN De-hui;ZHU Yu-qian;WANG Ling-yun;QI Meng;ZHENG Hai-xue(College of Animal Science,Tarim University,Aral 843300,China;National Foot-and-Mouth Disease Reference Laboratory,Key Laboratory of Animal and Poultry Virology,Ministry of Agriculture and Rural Affairs,State Key Laboratory of Veterinary Etiological Biology,Lanzhou Veterinary Research Institute of Chinese Academy of Agriculture Science,Lanzhou 730046,China)
出处
《中国兽医科学》
CAS
CSCD
北大核心
2022年第4期410-416,共7页
Chinese Veterinary Science
基金
国家重点研发计划项目(2021YFD1800100)
国家自然科学基金专项(31941002)
甘肃省重大科技专项(20ZD7NA006-01)
中国农业科学院所级重点任务项目(CAAS-ASTIPJBGS-20210101)。
关键词
非洲猪瘟病毒
T细胞表位
筛选
蛋白
抗原性
African swine fever virus
T cell epitope
screening
protein
antigenicity
