Cellular markers of T cell activation, exhaustion and senescence in HIV– and HIV+ post-menopausal women.

Expression of activation (CD38, HLA-DR, Ki-67), exhaustion (PD-1) and senescence (CD28, CD57, CD127) markers was evaluated by flow cytometry in live CD4 and CD8 T cells. Cryopreserved PBMC were thawed and rested overnight before staining with ViViD and monoclonal antibodies and subsequent acquisition on a flow cytometer. Gating strategy for the phenotypic analysis of T cells was performed as follows: Lymphocytes were gated based on forward and side scatter, and gates for exclusion of singlets and dead cells (ViViD+ events) were drawn. Statistical differences between groups were analyzed by Student t-test. Significant P values are shown in bold.本研究通过流式细胞术（flow cytometry）对活的CD4和CD8 T细胞中的活化标志物（CD38、HLA-DR、Ki-67）、耗竭标志物（PD-1）及衰老标志物（CD28、CD57、CD127）的表达水平进行了检测。将冻存的外周血单个核细胞（PBMC, peripheral blood mononuclear cell）解冻后过夜静置，随后使用ViViD染料与单克隆抗体进行染色，最终在流式细胞仪上完成样本数据采集。T细胞表型分析的设门策略如下：基于前向散射光与侧向散射光圈选淋巴细胞群，再设置门控以排除黏连体细胞与死细胞（ViViD阳性事件）。组间统计学差异采用两独立样本t检验（Student t-test）进行分析，显著性P值以粗体标注。