破膜剂对流式细胞术检测Th1/Th2/Th17细胞频率的影响  

The effects of Fix&Perm reagents on flow cytometry detection for Th1/Th2/Th17 cell subsets

作　　者：周春保[1] 史继静[1] 王立峰[1] 李元元[1] 张学秀[1] 袁今虹[1] 张纪元[1] 徐若男[1] 金磊[1] 施明[1] 王福生[1] 

机构地区：[1]中国人民解放军第三○二医院感染性疾病诊治与研究中心,北京100039

出　　处：《免疫学杂志》2016年第2期181-184,共4页Immunological Journal

基　　金：国家自然科学基金(81172779;81470837)

摘　　要：目的探讨不同破膜剂对流式细胞术检测Th1/Th2/Th17细胞频率的影响。方法以健康人外周血为研究对象,使用多参数流式细胞仪分析,比较3种不同破膜剂(常规改进的破膜剂、eBioscience公司破膜剂、BD公司破膜剂)对CD4^+T淋巴细胞内干扰素-γ(IFN-γ)、白介素-4(IL-4)和白介素-17(IL-17)检测结果的影响。结果在相同条件下,eBioscience公司破膜剂检测到的IL-4^+CD4^+T细胞频率显著高于其它2组(P<0.05);改进的破膜剂组(GJ)IL-17^+CD4^+T细胞频率明显高于其它组(P<0.05);但IFN-γ^+CD4^+T细胞的频率在3组间无统计学差异(P>0.05)。结论 3种破膜剂破膜后均可检测到IFN-γ^+CD4^+(Th1)、IL-4^+CD4^+(Th2)、IL-17^+CD4^+(Th17)T细胞,但不同破膜剂对不同类型Th细胞的检出频率有影响,因此操作过程中应根据不同的需求选择合适的破膜剂。This study designed to explore the effects of different Fix＆Perm reagents on detection of Th1/Th2/ Th17 cell subsets by flow cytometry. Peripheral blood mononuclear cells were collected from health adults, and the percentages of IFN-γ＋ Th cells, IL-17＋ Th cells and IL-4＋ Th cells were analyzed by muhiparameter flow cytometry using three kinds of Fix＆Perm reagents （the improved Fix＆Perm reagent from our Lab, Transcription Factor Staining Buffer Set from eBioscience, Cytofix/CytopermTM Plus Fixation/Permeabilization Kit from BD）. Compared with the improved Fix＆Perm reagent from our Lab and Cytofix/CytopermTM Plus Fixation/Permeabilization Kit from BD, the percentage of IL-4＋ Th cells stained by Fix＆Perm reagent from eBioscience were significant higher （P〈 0.05）; the percentage of IL-17＋ Th cells stained by the improved Fix＆Perm reagent from our Lab were the highestone among three groups （P〈0.05）; there was no significant difference in the percentages of IFN-γ＋ Th cells stained by the three Fix＆Perm reagents （P〉0.05）. The results revealed that IFN-γ＋ Th, IL-4＋Th and IL-17＋ Th cells could be detected by flow cytometry using the three Fix＆Perm reagents, but different Fix＆Perm reagents may affected the detection efficiency. So we should choose the optimized Fix＆Perm reagent according to experimental purpose.

关 键 词：流式细胞术 破膜剂 细胞因子 

分 类 号：R446.11[医药卫生—诊断学]