Single-Cell Analysis of HIV Latency Reversal

项目摘要 艾滋病毒在长寿宿主中的持续存在导致需要终身抗逆转录病毒治疗（ART）。 由于这些储层的潜在数量相对较少，因此对这些储层的准确量化具有挑战性。 一个人体内被感染的细胞用于HIV储库定量的最佳测定应该是高通量的， 灵敏，成本效益高，理想情况下能够量化HIV感染细胞的频率。电流 测量这些储库的大小的测定受到许多缺点的阻碍， 确定感染细胞数量，需要大量样品，以及成本和劳动密集型 某些化验。一种单细胞检测方法，可以量化具有活跃HIV转录的感染细胞的数量 将提供前所未有的清晰度，对艾滋病毒水库的状态，潜伏期的影响， 逆转剂，并将有重要意义的机制和基本发病机制的研究， 好.我们最近验证了一种用于检测HIV表达细胞频率的单细胞平台。 使用这种检测方法，我们建议评估一系列潜伏期逆转剂对HIV的有效性 重新激活，并探讨其差异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.