四聚体技术检测人外周血巨细胞病毒抗原特异性CTL  

Detection of the cytomegalovirus antigen-specific cytotoxic T lymphocytes in human peripheral blood mononuclear cells by using MHC class I tetramer technique

作　　者：李艳萍[1] 徐惠绵[1] 喻卫红[2] Xiaoning Xu 

机构地区：[1]中国医科大学附属第一医院肿瘤外科,沈阳110001 [2]辽宁省肿瘤医院研究所,沈阳110042 [3]牛津大学分子医学研究中心,英国牛津OX39DS

出　　处：《现代免疫学》2007年第3期248-252,共5页Current Immunology

基　　金：国家留学基金委出国留学基金(21921035)

摘　　要：为深入了解抗病毒免疫机制,探索人在健康状态及巨细胞病毒(CMV)急性感染时抗原特异性细胞毒T淋巴细胞(CTL)数量的动态变化。以CMV抗原肽、HLA-A*0201重链和轻链制备CMV四聚体;分离并检测12名健康被检者外周血单个核细胞(PBMC)中CMV抗原特异CTL数量;PBMC体外培养建立CTL细胞系,于末次刺激的不同时相进行四聚体染色,流式细胞仪(FACS)检测抗原特异CTL数量。结果发现9名被检者PBMC中均检出抗原特异CTL;细胞系中CMV特异性CTL数量急剧增多,细胞系与PBMC相比,差异有显著的统计学意义(P<0.001);研究结果提示,9名被检者感染过CMV,血液中存在少量免疫记忆性T细胞,当再次遭遇同一抗原后发生克隆扩增。To investigate the dynamic changes in the quantity of antigen-specific cytotoxic T lymphocytes （CTL） in healthy individuals and acute infection with cytomegalovirus（CMV） for a better understanding on the anti-viral immune mechanism, the CMV-tetramer was prepared by CMV antigen peptide together with heavy and light chain of HLA-A ＊ 0201. The peripheral blood mononuclear cells （PBMC） were isolated from blood of 12 healthy donors and cultivated ex vitro to set up the antigenspecific CTL lines and the quantity of antigen-specific CTL was determined by FACS using the CMV tetramer. It was found that low but detectable level of the CMV-specific CD8＋ T cells could be detected in PBMC of 9 donors by FACS analysis. How ever, for the CTL line, the quantity of CMV-specific CTL increased obviously with significant difference in comparison with that of PBMC （P〈0. 001）. It is evident that 9 donors had already suffered from CMV infection with the presence of small amount of memory T cells which can specifically respond to reinfections.

关 键 词：四聚体 巨细胞病毒 细胞毒T细胞 

分 类 号：R373[医药卫生—病原生物学]