Beliakova-Bethell Nadejda, Hezareh Marjan, Wong Joseph K, Strain Matthew C, Lewinski Mary K, Richman Douglas D, Spina Celsa A
VA San Diego Healthcare System, 3350 La Jolla Village Dr., San Diego, CA 92161, USA; University of California San Diego, 9500 Gilman Dr., La Jolla, CA 92093, USA.
University of California San Diego, 9500 Gilman Dr., La Jolla, CA 92093, USA.
Virology. 2017 Aug;508:127-133. doi: 10.1016/j.virol.2017.05.008.
Quantification of cell-associated replication-competent HIV, in blood samples from patients with undetectable plasma viremia, requires specialized culture conditions that include exogenous pan T cell stimulation. Different research groups have used several stimuli for this purpose; however, the relative efficacies of these T cell stimuli to induce productive HIV replication from latently infected cells ex vivo have not been systematically evaluated. To this end, we compared four commonly used T cell stimuli: 1) irradiated allogeneic cells plus phytohaemagglutinin (PHA); 2) PHA alone; 3) phorbol myristate acetate plus Ionomycin; and 4) immobilized αCD3 plus αCD28 antibodies. End-point dilutions of patient CD4 T cells were performed, using virion RNA production to quantify HIV induction. Our results demonstrated that these activation approaches were not equivalent and that antibody cross-linking of CD3 and CD28 membrane receptors was the most effective means to activate HIV replication from a resting cell state, closely followed by stimulation with irradiated allogeneic cells plus PHA.
对血浆病毒血症检测不到的患者血样中与细胞相关的具有复制能力的HIV进行定量分析，需要专门的培养条件，包括外源性全T细胞刺激。不同的研究小组为此使用了多种刺激物；然而，这些T细胞刺激物在体外从潜伏感染细胞诱导产生有 productive HIV复制的相对效力尚未得到系统评估。为此，我们比较了四种常用的T细胞刺激物：1）辐照的异基因细胞加植物血凝素（PHA）；2）单独使用PHA；3）佛波酯肉豆蔻酸酯乙酸盐加离子霉素；4）固定化的αCD3加αCD28抗体。使用病毒体RNA产生来定量HIV诱导，对患者CD4 T细胞进行终点稀释。我们的结果表明，这些激活方法并不等效，CD3和CD28膜受体的抗体交联是从静止细胞状态激活HIV复制的最有效手段，紧随其后的是用辐照的异基因细胞加PHA进行刺激。
