Vγ9Vδ2T细胞亚群的建立与功能分析  

Establishment and functional analysis of Vγ9Vδ2T cell subpopulation

作　　者：邱潮林 周晓薇[2] 胡朝英[3] 谢秋华 胡锦辉 赵蓉[2] 钱柳[2] 张冬青[2] 

机构地区：[1]上海浦东新区公利医院检验科,上海200135 [2]上海交通大学医学院上海市免疫学研究所,上海200025 [3]上海市徐汇区中心医院检验科,上海200031

出　　处：《诊断学理论与实践》2013年第2期210-215,共6页Journal of Diagnostics Concepts & Practice

基　　金：国家自然科学基金项目(31270963);上海市科委面上项目(10ZR1426100)

摘　　要：目的:建立γδT细胞的一个亚群(Vγ9Vδ2 T细胞亚群),分析其免疫学相关功能。方法:对健康人外周血单个核细胞进行磁珠分选,获得富含γδT细胞的细胞群,用异戊烯焦磷酸(isopentenyl pyrophosphate,IPP)、白细胞介素2(IL-2)刺激γδT细胞增殖。通过流式细胞仪检测分析Vγ9Vδ2 T细胞(γδT细胞主要亚群)在IPP刺激后的分化趋势及表面标志,分离出效应型Vγ9Vδ2 T细胞,对其抗原递呈功能进行研究,并分析不同表型的Vγ9Vδ2 T细胞的分泌格局。结果:Vγ9Vδ2 T细胞在IPP作用下可被激活,不断增殖并分化为效应记忆型(TEM、CD45RA-CD27-γδT),该细胞群高表达HLA-DR和CD80/86等分子,呈现出专职性抗原递呈细胞的表型特征;此群效应记忆型Vγ9Vδ2 T细胞能有效递呈PPD、TT等可溶性抗原给CD4+T细胞,具有与B细胞相当的抗原递呈功能。实验还发现,不同表型的Vγ9Vδ2 T细胞具有不同的细胞因子分泌格局,新鲜分离的健康人外周血Vγ9Vδ2 T细胞(非效应型)以分泌IFN-γ为主,而效应记忆型Vγ9Vδ2 T细胞在产生IFN-γ的同时能分泌IL-17。结论:Vγ9Vδ2 T细胞亚群能在IPP的作用下激活、增殖,并分化为具有抗原递呈细胞功能的效应记忆型细胞,其分泌IFN-γ的同时可分泌IL-17。Objective To establish a subpopulation of γδ T cell （Vγ9Vδ2T cell subpopulation） and study its immune related function. Methods A cell population rich in γδ T cells was obtained by magnetic beads sorting method from peripheral mononuclear cells of healthy people. IPP （isopentenyl pyrophosphate） and IL-2 were used to stimulate the proliferation of γδ T cell. Flow cytometry was used to analyze the differentiation trends and surface marker of Vγ9Vδ2T cells after IPP stimulation. Effector type Vγ9Vδ2T cell was separated and its antigen presenting function was studied, and the cytokine secretion pattern of different phenotypic Vγ9Vδ2T cells was analyzed. Results Vγ9Vδ2T cells （the predominant subtype of γδ T cells in peripheral blood） were activated by IPP to continuously proliferate and differentiate into effector memory type （TEM, CD45RA, CDγδ T）. The effeetor memory Vγ9Vδ2T cells expressed HLA-DR and CD80/86 and exhibited the phenotypic characteristics of antigen-presenting cells. These Vγ9Vδ2T cells could present soluble antigens （PPD and TI） to CD4＋T cells, having the corresponding antigen-presenting effect as B cells. Vγ9Vδ2T cells of different phenotypes showed different cytokine secretion patterns. Non-effector Vγ9Vδ2T cells secreted predominantly IFN-γ whereas effeetor memory Vγ9Vδ2T cells secreted simultaneously IFN-γ and IL-17. Conclusions： Vγ9Vδ2T cell could be activated by IPP stimulation, and proliferated and differentiated into effector memory type Vγ9Vδ2T cells with antigen- presenting function, not only secreting IFN-γ but also IL-17. This study provided the theoretical and experimental data for further study of γδ T cells and its correlation with clinical diseases.

关 键 词：γδ T细胞 效应记忆型Vγ9Vδ2T细胞 抗原递呈作用 

分 类 号：R446.63[医药卫生—诊断学]