三色免疫荧光标记法检测外周血Th细胞亚群  

Three Color Immuno-fluorescence Detection of Th-Subsets in Peripheral Blood

作　　者：叶卫东[1] 张循善[2] 翟志敏[3] 连莉[2] 孙小梅[3] 李庆[3] 

机构地区：[1]武警安徽总队医院检验科,合肥230041 [2]安徽医科大学第一附属医院 [3]安徽省立医院中心实验室

出　　处：《临床输血与检验》2008年第2期119-121,共3页Journal of Clinical Transfusion and Laboratory Medicine

基　　金：安徽省教育厅科研基金项目(No2001kj147)资助

摘　　要：目的建立一种流式细胞仪三色法检测外周血Th细胞亚群的方法。方法外周全血用RPMI1640培养液和终浓度为25ng/ml的PMA、1μg/ml的Ionomycin、20ng/ml的Monensin培养2h,固定和破膜后,加入荧光标记的CD4、IFN-γ和IL-4抗体,应用流式细胞仪进行检测。结果CD4+细胞与CD4-细胞能明显区分,12份标本Th0、Th1和Th2分别为(2.46±0.97)%、(12.77±4.01)%和(1.82±0.81)%。结论三色免疫荧光标记法可以同时快速准确地测定外周血Th0、Th1和Th2细胞亚群。Objective To establish a three-color method for detecting Th subsets in peripheral blood by flow cytometry. Methods Peripheral whole blood were co-cultured with RPMI1640,25 ng/ml PMA, 1 μg/ml ionomycin and 20 ng/ml monensin for 2 hours, fixed and permeabilized,then fluorescein-labeled antibody of CD4,IFN-γ and IL-4 were added. The cells containing corresponding antigen were detected by flow cytometry. Results CD4^＋ cells and CD4^-cells could be discriminated by CD4 molecular of cell surface. Th0 ,Th1 and Th2 of 12 samples were （2.46±0. 97）%, （12.77±4.01）% and （1.82±0. 81）% respectively. Conclusion This method can detect Th0,Th1and Th2 simultaneously, quickly and exactly.

关 键 词：流式细胞仪 三色免疫荧光 T辅助细胞亚群 

分 类 号：R392.11[医药卫生—免疫学]