Role of HIV Rev in Reactivation from Latency

项目摘要/摘要 治愈艾滋病毒的努力继续受到持续存在的诱导性前病毒的阻碍。 潜伏感染的细胞。如果构成前病毒储存库的细胞能够被诱导活跃地产生病毒， 然后，可能会通过天然免疫杀死或外在疗法来清除病毒。努力 到目前为止，由于不能可靠地刺激整个潜在储集层 这一许可是可以实现的。一种可能导致潜伏细胞不理想的潜在机制 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功能活性在以下几个方面差异很大 REV/RRE活性调节病毒复制，而不是起开关作用。 在重新激活时，潜伏感染细胞中功能较不活跃的REV/RRE对中的前病毒将是 预计会减少许多病毒蛋白和病毒颗粒的产生，同时继续产生 正常水平的免疫调节病毒Nef蛋白。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，活动可能更难清除。 这项提议中的实验旨在检验这一假设。Subaims1a和1b将利用 已建立延迟模型和一组已将REV/RRE活性直接更改为 评估使用各种潜伏再活化剂重新激活潜伏感染细胞对病毒激活的影响 以及成长的结果。Subaim1c将测量从中国分离的具有复制能力的病毒的REV/RRE功能 病人的潜伏细胞。这些实验将确定REV/RRE功能与序列相比有何不同 从新诊断的患者的外周血中提取，以及是否有任何选择与 关于REV/RRE功能活动的水平。特定目标2将测量潜伏期重新激活的CTL杀伤 直接确定REV/RRE活性变化如何影响细胞对免疫的敏感性 淘汰赛。 对REV-RRE调节系统如何在病毒潜伏期发挥作用的更好的理解可能是 这是理解为什么“先踢后杀”的治疗策略迄今不成功的关键。此信息 反过来，可以通过调节潜伏病毒的REV-RRE活性来优化“踢”。建议数 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战略，并可能提供至关重要的 推进这一迄今难以实现的目标的知识。

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.