Phenotypic analysis of the unstimulated in vivo HIV CD4 T cell reservoir

The latent reservoir is a main barrier for curing HIV. But because latently-infected cells cannot be phenotyped directly, the features of the in vivo reservoir have remained elusive. Here, we describe a method that leverages high-dimensional phenotyping using CyTOF to trace latently-infected cells reactivated ex vivo to their original pre-activation states. Our results suggest that contrary to common assumptions, the reservoir is not randomly distributed among cell subsets, and is remarkably conserved between individuals. However, reservoir composition differs between tissues and blood, as do cells successfully reactivated by different latency reversing agents. By selecting 8-10 of our 39 original CyTOF markers, we were able to isolate highly purified populations of unstimulated in vivo latent cells. These purified populations were highly enriched for replication-competent and intact provirus, transcribed HIV, and displayed clonal expansion. The ability to isolate unstimulated latent ce...

潜伏储库是治愈人类免疫缺陷病毒（HIV）感染的主要障碍。由于无法直接对潜伏感染细胞进行表型分型，体内潜伏储库的特征始终难以阐明。本研究报道一种方法，借助质谱流式细胞技术（CyTOF）开展的高维表型分型，可将体外激活的潜伏感染细胞追溯至其原始激活前状态。研究结果显示，与普遍假设相悖的是，潜伏储库并非随机分布于各类细胞亚群中，且在不同个体间呈现显著保守性。不过，储库的组成在不同组织与血液之间存在差异，不同潜伏期逆转剂成功激活的细胞亦是如此。通过从39个原始CyTOF标志物中筛选8-10个，我们得以分离得到高纯度的未受刺激体内潜伏细胞群体。这些纯化后的群体高度富集具有复制能力的完整前病毒、转录型HIV，并展现出克隆扩增特性。分离未受刺激潜伏细胞的能力……