HIV persistence in vivo

该项目的目的是帮助阐明艾滋病毒如何在体内持续存在，从而帮助开发新的艾滋病毒治疗方法。这项工作的重点是一小群CD4 T细胞，它们含有具有复制能力的艾滋病毒，即使在有效的抗逆转录病毒治疗（ART）的环境中，它们的水平也保持在接近稳定的水平。尽管这些细胞被认为是“潜在的储存库”，但维持其数量的细胞过程和潜在的基因表达模式仍然不完全确定。实际上，感染性病毒在细胞间的持续传播、接受抗逆转录病毒治疗前被感染细胞的增殖以及受感染细胞的长期静止（“潜伏期”）都可能成为有效抗逆转录病毒治疗期间治愈的障碍。区分这些机制对体内HIV治愈障碍的贡献将是至关重要的，因为旨在对抗每种单独机制的干预措施可能无法影响甚至恶化其他机制。

The purpose of this project is to help elucidate how HIV persists in vivo and thus to aid the development of novel HIV cure-directed therapies. The focus of the work is a small population of CD4 T cells that contain replication-competent HIV and persist at nearly stable levels even in the setting of effective antiretroviral therapy (ART). Although these cells have been considered to represent a "latent reservoir," the cellular processes and underlying gene expression patterns that maintain their numbers remain incompletely defined. Indeed, it remains possible that the ongoing cell-to-cell spread of infectious virus, the proliferation of cells infected before institution of ART, and the long-term quiescence ("latency") of infected cells all present barriers to cure during effective ART. Distinguishing the contributions of these mechanisms to the barrier to HIV cure in vivo will be critical because interventions designed to antagonize each individual mechanism may fail to affect or even worsen the others. Unfortunately, distinguishing the contributions of these mechanisms in blood and tissue samples from HIV-infected study participants has been extremely challenging. A major reason for this is that the CD4 T cells that contain HIV in vivo often lack expression of virus- or host-encoded markers. This prevents identification and isolation of highly pure populations of cells for the types of comprehensive downstream analysis that might reveal transcriptomic, epigenetic, or proteomic signatures of ongoing virus replication, cellular proliferation, or cellular latency. In this project, we are developing two complementary approaches to overcoming this technical challenge: a reverse approach in which phenotypic CD4 T cell subsets are sorted by flow cytometry for downstream characterization as hosts for the virus, and a forward approach in which cells containing HIV genomic DNA are isolated for comprehensive analysis following detection in water-in-oil emulsions.