Role of HIV Rev in Reactivation from Latency

项目总结/摘要 艾滋病病毒感染者体内持续存在可诱导的前病毒， 潜伏感染细胞如果可以诱导构成前病毒库的细胞主动产生病毒， 那么储库可能通过天然免疫杀伤或外源性治疗而被清除。努力 迄今为止，在这一方向上，由于不能可靠地刺激整个潜在储层， 这个间隙是可以实现的。一种潜在的机制，可能有助于次优的潜伏细胞 激活和清除是HIV Rev/RRE调节轴的变化。 我们实验室的最新数据表明，Rev/RRE功能活性在不同的细胞中差异很大。 主要分离株，而不是作为一个开关，Rev/RRE活性调节病毒复制。 在重新激活后，具有较低功能活性Rev/RRE对的潜伏感染细胞中的前病毒将被激活。 预计将减少许多病毒蛋白和病毒颗粒的生产，同时继续生产 免疫调节病毒Nef蛋白的正常水平。含有低Rev/RRE的前病毒的细胞 因此，在所谓的“踢-杀”治疗策略中，活性可能更难以清除。 本提案中的实验旨在验证这一假设。子项目1a和1b将利用 建立的潜伏期模型和一组改变Rev/RRE活性的工程病毒， 评估用各种潜伏再活化剂再活化潜伏感染细胞如何影响病毒活化 和增长。Subaim 1c将测量分离自以下病毒的复制能力病毒中的Rev/RRE功能： 病人的潜伏细胞这些实验将确定Rev/RRE功能与序列相比如何变化 来自新诊断患者的外周血，以及是否有任何选择， 关于Rev/RRE功能活动的水平。特异性目标2将测量潜伏再活化的CTL杀伤。 细胞，以直接确定Rev/RRE活性变化如何影响细胞对免疫应答的敏感性。 淘汰 对Rev-RRE调节系统在病毒潜伏期中如何发挥作用的更好理解可能是其中之一 这是理解为什么迄今为止“踢死”治疗策略不成功的关键。这些信息 可以反过来用于通过调节潜伏病毒的Rev-RRE活性来优化“踢”。拟议 因此，这项研究直接关系到迄今为止最有希望治愈艾滋病毒的策略，并可能提供关键的 知识来推进这一迄今为止难以捉摸的目标。

Project Summary/Abstract Efforts to achieve a cure for HIV continue to be stymied by the persistence of inducible proviruses in latently infected cells. If the cells constituting the proviral reservoir can be induced to actively produce virus, then the reservoir could potentially be cleared by means of native immune killing or extrinsic therapies. Efforts in this direction to date have been hampered by an inability to reliably stimulate the entire latent reservoir such that this clearance can be achieved. One potential mechanism that could contribute to suboptimal latent cell activation and clearance is variation in the HIV Rev/RRE regulatory axis. Recent data from our laboratory have indicated that Rev/RRE functional activity varies greatly among primary isolates and that rather than acting as an on-off switch, Rev/RRE activity modulates viral replication. Upon re-activation, proviruses in latently infected cells with less functionally active Rev/RRE pairs would be expected to have decreased production of many viral proteins and virus particles, while continuing to produce normal levels of the immune modulatory viral Nef protein. Cells containing proviruses with low Rev/RRE activity may therefore be more difficult to clear in the so-called “kick-and-kill” strategies for cure. Experiments in this proposal are designed to test this hypothesis. Subaims1a and 1b will utilize an established latency model and a set of engineered viruses that have altered Rev/RRE activity to directly assess how reactivation of latently infected cells with various latency reactivating agents affects viral activation and outgrowth. Subaim1c will measure Rev/RRE function in replication competent viruses isolated from patients' latent cells. These experiments will determine how Rev/RRE function varies compared to sequences derived from the peripheral blood of newly diagnosed patients, and whether there has been any selection with respect to levels of Rev/RRE functional activity. Specific Aim 2 will measure CTL killing of latency-reactivated cells to directly determine how Rev/RRE activity variation influences the susceptibility of the cells to immune elimination. Improved understanding of how the Rev-RRE regulatory system functions in viral latency may be one key to understanding why “kick-and-kill” cure strategies have been unsuccessful to date. This information could in turn be used to optimize the “kick” by modulating the Rev-RRE activity of latent virus. The proposed study thus has direct bearing on the most promising strategy to date to cure HIV and could provide crucial knowledge to advance this thus-far elusive goal.