Single-Cell Analysis of HIV Latency Reversal

项目总结 艾滋病毒在长寿的宿主体内持续存在，导致需要终生抗逆转录病毒疗法(ART)。 由于潜在储集层的数量相对较少，准确量化这些储集层是具有挑战性的。 一个人体内被感染的细胞。HIV储备库定量的最佳方法应该是高通量的， 灵敏、成本效益高，理想情况下能够量化艾滋病毒感染细胞的频率。当前 测量这些储集层大小的分析受到一些缺点的阻碍，包括无法 为了确定感染细胞的数量，需要大量的样本，以及成本和劳动密集型的性质 某些化验结果。一种可以量化艾滋病毒转录活跃的感染细胞数量的单细胞检测方法 将对艾滋病毒宿主的状态、潜伏期的影响提供前所未有的清晰程度- 逆转剂，并将对机制和基础发病机制研究具有重要意义 井。我们最近验证了一个单细胞平台，用于检测表达艾滋病毒的细胞的频率。 利用这项试验，我们建议评估一系列潜伏期逆转药物对艾滋病毒的有效性。 并探索其不同效应背后可能的潜在机制。

PROJECT SUMMARY The persistence of HIV within long-lived reservoirs results in the need for life-long antiretroviral therapy (ART). The accurate quantification of these reservoirs is challenging due to the relatively small number of latently infected cells within an individual. The optimal assay for HIV reservoir quantification should be high-throughput, sensitive, cost-efficient, and would ideally be able to quantify the frequency of HIV-infected cells. Current assays to measure the size of these reservoirs are hindered by a number of drawbacks, including the inability to determine infected cell numbers, the need for large sample volumes, and the cost and labor-intensive nature of certain assays. A single-cell assay that can quantify the number of infected cells with active HIV transcription would provide an unprecedented level of clarity on the state of the HIV reservoir, the effects of latency- reversing agents, and would have important implications for mechanistic and basic pathogenesis studies as well. We have recently validated a single-cell platform for detecting the frequency of HIV-expressing cells. Using this assay, we propose to evaluate the effectiveness of a range of latency-reversing agents for HIV reactivation and to explore a possible underlying mechanism behind their differential effects.