CFSE标记的淋巴细胞增殖实验检测方法的建立  

CFSE labeled cells for the detection of lymphocyte proliferation

作　　者：孙尔维[1] 袁小澎[2] 罗宇维[2] 陈忠华 

机构地区：[1]华中科技大学同济医学院附属同济医院器官移植研究所 [2]南方医科大学珠江医院移植免疫研究所,广州510282

出　　处：《广东医学》2008年第10期1626-1628,共3页Guangdong Medical Journal

基　　金：国家自然科学基金资助项目(编号:30571757);广东省自然科学基金团队项目(编号:5200513);广州市科技计划项目(编号:2007Z3-E0051)

摘　　要：目的建立基于流式细胞术和活细胞荧光染料CFSE的淋巴细胞增殖、单向混合淋巴细胞实验的免疫学检测方法。方法分离单个核细胞,将反应细胞经CFSE染色后,分别加入不同的淋巴细胞增殖刺激物或与灭活的异基因淋巴细胞共培养。培养后细胞经流式细胞术检测标记细胞是否增殖,应用CD4-PE抗体检测增殖细胞中CD4阳性T细胞亚群比例,ModFit软件分析增殖动力模型。计算增殖CDI或PI指数。结果基于CFSE建立的淋巴细胞增殖实验,经不同刺激物或异基因抗原刺激后增殖指数明显升高,使用CD4-PE抗体进行再次标记后可以对增殖实验中CD4阳性淋巴细胞亚群的增殖进行评估。结论成功建立了基于CFSE标记淋巴细胞的增殖实验检测方法,该方法简单、实用、灵敏度高,可以取代3H或MTT用于不同条件、不同免疫刺激情况下淋巴细胞增值的检测或评估。Objective To establish an immunological methop to detect lymphocyte proliferation with CFSE labeled cells. Methods PBMCs were isolated from normal subjects and labeled with CFSE, and were subseqently stimulated for 5d with PHA, ConA and allogeneic splenocytes respectively. The CD4^＋ phenotypes of proliferating lymphocytes were determined by flow cytometry with specific antibodies. The proliferation dynamics of CFSE labeled cells was detected by flow cytometry and analyzed with the ModFit software. Results Lymphocytes could be activated by PHA, ConA or allogeneic antigen. Proliferation of different lymphocyte subses can be determined by labeling with specific lymphocytes. Conclusion With assistance of CFSE labeling, flow cytometry can be used to obtain more detailed information on the distinctive response of different T cell subsets and on the dynamics of a specific T cell subset.

关 键 词：羧基荧光素乙酰乙酸琥珀酰亚胺酯 增殖 流式细胞术 

分 类 号：R977.9[医药卫生—药品] R392.11[医药卫生—药学]