HIV/AIDS感染者体外诱导的γδT细胞亚群的特点

Characteristics of γδ T cell subsets induced from peripheral blood mononuclear cells of HIV /AIDS patients in vitro

目的:了解HIV/AIDS感染者外周血中γδT细胞亚群的特点以及体外诱导后γδT细胞的增殖情况和Vδ1、Vδ2亚群的变化,为γδT细胞体外扩增方法的建立打下基础。方法:流式细胞术(FCM)检测25例HIV/AIDS感染者(HIV组)和31例健康对照者(HC组)外周血中的γδT细胞、Vδ1亚群、Vδ2亚群的频率和绝对值;选择两组中各10例外周血单个核细胞(PBMC),用anti-γδTCR单克隆抗体(mAb)和IL-2在体外诱导培养14 d,在0 d、7 d、14 d分别进行细胞计数和FCM检测,分析比较γδT细胞亚群的比例和培养情况。结果:HIV组外周血中γδT细胞、Vδ2亚群百分比和绝对值都显著低于HC组,而Vδ1亚群百分比和绝对值显著高于HC组;体外诱导培养14 d时,HC组总细胞数量增加3倍,HIV组总细胞数量稍有增加;HC组的γδT细胞比例可达到80%以上,其中Vδ2亚群可以增加到65%左右,而HIV组的γδT细胞比例达35%左右,Vδ2亚群比例仅增加到17%左右。结论:HIV/AIDS感染者外周血中γδT细胞比例显著降低,Vδ1/Vδ2比值倒置;用anti-γδTCR mAb和IL-2在体外诱导HIV/AIDS感染者的Vδ2亚群扩增效果不佳,不能逆转Vδ1/Vδ2倒置现象。应进一步寻找更加有效的诱导培养方法。

AIM： To investigate the characteristics of γδ T cell subsets in peripheral blood and Vδ1 and Vδ2 T cell subsets proliferation after induction in vitro,and to provide experimental basis for γδ T cells expansion methods in vitro.METHODS： Percentages of γδ T cells,Vδ1 and Vδ2 T cells in CD3 T cells from 25 cases of HIV /AIDS patients（HIV group） and 31 cases of healthy adults as control（HC group） were investigated with flow cytometry（FCM）;and 10 cases peripheral blood mononuclear cells（PBMCs） from each group were induced and cultured for 14 d by using antiγδ TCR mAb and IL-2,then the cell amounts were counted,γδ T cells and Vδ1,Vδ2 T cells were detected and analyzed with flow cytometry（FCM） at 0 d,7 d and 14 d.RESULTS： The percentages and absolute counts of γδ T cells and V δ2 T cells in HIV group were significantly lower than those in HC group,but those of Vδ1 subsets were significantly higher than those of HC group.After culture for 14 day,the total cell amount of HC group expanded almost 3 times,but those of HIV group expanded a little;The percentages of γδ T cells in HC group were above 80%,but those in HIV group were only about 35%;the percentages of Vδ2 T cells in HC group were about 65%,but those in HIV group were only about 17%.CONCLUSION： The proportion of γδ T cells in peripheral blood of HIV /AIDS patients decreased significantly,and Vδ1 /Vδ2 ratio was inversed;the expansion effect was not so good to use antiγδ TCR mAb and IL-2 to induce γδ T cells and Vδ2 subsets of HIV /AIDS,and the Vδ1 /Vδ2 ratio inversion could not reverse.More efficient culture methods should be explored.