一种CD3^＋ T细胞细胞内CD69分子的检测方法  

A method for detecting intracellular CD69 molecules in CD3^+ T cells

作　　者：王静[1] 王一任 王克强 

机构地区：[1]泰安市中医医院检验科,山东泰安271000 [2]青岛大学医学部,山东青岛266000

出　　处：《泰山医学院学报》2017年第12期1337-1339,共3页Journal of Taishan Medical College

基　　金：山东省保健科技协会科学技术课题(2016BJ0012)

摘　　要：目的建立一种人外周血CD3^+ T细胞细胞内CD69的检测方法。方法分离获取健康人PBMC,用PMA^+ IM(佛波醇酯^+ 离子霉素)刺激PBMC,用蛋白转运抑制剂(Brefeldin A,BFA)阻断细胞内因子的分泌,用皂角素(Saponin)破膜,流式细胞术检测CD3^+ T细胞胞内CD69阳性率,以确定最佳的破膜时间。结果最佳的破膜时间为15 min,CD3^+ T细胞胞内CD69表达的阳性率较高,可达87.82%。结论流式细胞术是一种检测CD3^+ T细胞内CD69的较好方法,这一方法也为CD3^+ T细胞内其他分子的检测分析奠定了方法学基础。Objective: To establish a method for detecting intracellular CD69 in human peripheral blood CD3^+ T cells.Methods: Healthy human PBMC was isolated,PBMC were stimulated by PMA and IM. Protein transport inhibitors( Brefeldin A,BFA) were used to block the secretion of cytokines. Saponin was used to destruct cell membrane. In order to find the best time of membrane destruction,we used flow cytometry to detect the positive rate of CD69 in CD3^+ T cells. Results:The best time of membrane destruction was 15 minutes. The positive rate of CD69 in CD3^+ T cells was higher,even up to87. 82%. Conclusion: Flow cytometry is a good method for the detection of intracellular CD69 in CD3^+ T cells,and flow cytometry also make a methodological foundation for the detection of other cytokines in CD3^+ T cells.

关 键 词：CD3^+T细胞 CD69分子 PMA+IM 流式细胞术 

分 类 号：R392-33[医药卫生—免疫学]