流式细胞术检测CFSE标记人T细胞亚群增殖反应  

Determination of the proliferation of CFSE labeled human T lymphocyte subpopulations by flow cytometry

作　　者：王敏[1] 张林杰[1] 何金生[1] 葛晓松[2] 陈勇[2] 李柏青[1] 

机构地区：[1]安徽医科大学免疫学教研室,合肥230001 [2]蚌埠医学院免疫学教研室,安徽蚌埠233003

出　　处：《免疫学杂志》2005年第5期423-426,共4页Immunological Journal

基　　金：国家自然科学基金资助项目(30070721)

摘　　要：目的探讨应用流式细胞术和活细胞荧光染料CFSE检测T淋巴细胞各亚群增殖反应的方法学。方法人外周血单个核细胞(PBMC)经CFSE染色后,分别用植物凝血素(PHA)、CD3mAb和结核杆菌抗原(Mtb-Ag)刺激,加IL-2扩增后,用流式细胞术检测活化增殖后T细胞各亚群的比例,并用ModFit软件分析各亚群的增殖动力模型。结果PHA和CD3mAb主要活化总T细胞,CD8+T细胞的增殖优于CD4+T细胞,但CD4+T细胞前4代细胞明显多于CD8+T细胞。Mtb-Ag主要刺激γδT细胞增殖。结论以CFSE标记淋巴细胞,结合流式细胞术可有效检测出不同T细胞亚群对不同刺激剂的增殖反应以及增殖动力学变化。Objective To evaluate the method of determining the proliferation of human T lymphocyte subpopulations by using flow cytometry and carboxyiluorescein diacetate succinimidyl ester（CFSE） labeling assay. Methods PBMC isolated from normal subjects were labeled with CFSE, and stimulated with PHA, CD3 mAb, and Mtb-Ag, respectively. Then, the stimulated PBMC was cultivated in medium containing IL-2. The phenotypes of proliferated lymphocytes were determined by flow cytometry. The dynamics of the proliferation of activated T cell subpopulations were analyzed with the ModFit software, Results Total T cells were mostly activated by PHA and CD3Ab, Different proliferations were observed between CD4^＋ T and CD8^＋ T eells, in which CD8^＋ T cells had greater reproduction, On the other hand, Mtb-Ag was mainly stimulated the reproduction of the γδT eells. Conclusion With assistance of CFSE labeling assay, the flow eytometry is able to obtain more detail information of proliferative response and dynamics of different T cell subpopulations after stimulated with different stimuli.

关 键 词：羧基荧光素乙酰乙酸琥珀酰亚胺酯 增殖 T细胞亚群 流式细胞术 

分 类 号：R392[医药卫生—免疫学]