Chen Wei-wei, Wu Xiao-li, Nie Wei-min, Xie Yang-xin, Wang Li, Zhao Min
Treatment and Research Center for Infectious Diseases, Beijing, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2012 Mar;28(3):285-7.
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.
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 anti-γδ 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.
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%.
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 anti-γδ 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.
研究外周血γδT细胞亚群的特征以及体外诱导后Vδ1和Vδ2T细胞亚群的增殖情况,为体外扩增γδT细胞的方法提供实验依据。
采用流式细胞术(FCM)检测25例艾滋病患者(HIV组)和31例健康成年人(HC组)外周血CD3⁺T细胞中γδT细胞、Vδ1和Vδ2T细胞的百分比;每组取10例外周血单个核细胞(PBMC),用抗γδTCR单克隆抗体和IL-2诱导培养14 d,分别于0 d、7 d和14 d进行细胞计数,并采用流式细胞术(FCM)检测分析γδT细胞以及Vδ1、Vδ2T细胞。
HIV组γδT细胞和Vδ2T细胞的百分比及绝对计数均显著低于HC组,而Vδ1亚群的百分比及绝对计数显著高于HC组。培养14 d后,HC组细胞总量扩增近3倍,而HIV组仅稍有扩增;HC组γδT细胞百分比高于80%,而HIV组仅约为35%;HC组Vδ2T细胞百分比约为65%,而HIV组仅约为17%。
艾滋病患者外周血γδT细胞比例显著降低,Vδ1/Vδ2比例倒置;采用抗γδTCR单克隆抗体和IL-2诱导艾滋病患者γδT细胞及Vδ2亚群的扩增效果不佳,且Vδ1/Vδ2比例倒置无法逆转,需探索更有效的培养方法。