多色流式细胞术检测HIV/AIDS患者CD4+T细胞亚群变化

Change of CD4 T cell subsets of HIV/AIDS patients by multi-color flow cytometry

目的以多色流式细胞术为基础,检测HIV/AIDS患者外周血单个核细胞中辅助性T细胞(helper T cell, Th)亚群的比例。方法用11种细胞表面抗体CD3、CD4、CD8、7-AAD、CD45RA、CCR7、CXCR5、CCR4、CCR6、CCR10、CXCR3建立多色流式方案,识别7种主要的Th,并对15例HIV/AIDS患者和15例健康对照外周血中Th的比例进行分析。结果根据多种趋化因子受体组合搭配,明确Th1、Th2、Th9、Th17、Th22、滤泡辅助性T细胞、ThG分析策略。通过检测分析发现,与健康对照相比,HIV/AIDS患者中Th2亚群比例增高,Th22亚群比例降低。结论建立的多色流式表面标志物检测方案可以方便快捷地检测外周血中Th的比例,所需样本量少,操作步骤简单且结果稳定,并能精准定量。

Objective To detect the proportion of CD4+ helper T cell(Th) subsets in peripheral blood monocyte cells of HIV/AIDS patients by multi-color flow cytometry. Methods The multi-color flow cytometry strategy was established by using 11 cell surface antibodies including CD3, CD4, CD8, 7-AAD, CD45 RA, CCR7, CXCR5, CCR4, CCR6, CCR10 and CXCR3. Seven major Th cells were recognized, and the proportion of Th in peripheral blood from 15 HIV/AIDS patients and 15 healthy controls was analyzed. Results Th1, Th2, Th9, Th17, Th22, T follicular helper cell and ThG analytical strategies were identified through the combination of different chemokine receptors. The detection results showed an elevated proportion of Th2 subset and decreased proportion of Th22 subset in HIV/AIDS patients, compared to healthy controls. Conclusions The established multi-color flow cytometric protocol by using surface markers can evaluate the proportions of multiple Th in peripheral blood conveniently and rapidly,which has small sample size, simple steps, stable results and precise quantitation.