Role of HIV Rev in Reactivation from Latency

HIV Rev 在潜伏期重新激活中的作用

• 批准号: 9534516
• 负责人: MARIE-LOUISE HAMMARSKJOLD
• 金额: $ 20.13万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-07-26 至 2020-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9534516
• 关键词: Affect; Anti-Retroviral Agents; Autologous; CD4 Positive T Lymphocytes; CD8-Positive T-Lymphocytes; Cells; Coculture Techniques; Data; Elements; Engineering; Goals; HIV; Highly Active Antiretroviral Therapy; Immune; Immune Evasion; Knowledge; Laboratories; Latent Virus; Lymphocyte; Measures; Memory; Modeling; Newly Diagnosed; Patients; Peptides; Pharmaceutical Preparations; Population; Predisposition; Process; Production; Proviruses; RNA Binding; Reporter; Research; Role; System; T-Lymphocyte; Testing; Variant; Viral; Viral Proteins; Viral Structural Proteins; Virion; Virus; Virus Activation; Virus Latency; Virus Replication; cytotoxic; design; experimental study; immune clearance; improved; nef Protein; peripheral blood; reactivation from latency; rev Genes

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.

项目概要/摘要 由于诱导型原病毒在体内的持续存在，治愈艾滋病毒的努力继续受到阻碍。 如果可以诱导构成原病毒库的细胞主动产生病毒， 那么可以通过天然免疫杀伤或外源疗法来清除储存库。 迄今为止，在这个方向上的努力因无法可靠地刺激整个潜在储层而受到阻碍，例如 可以实现这种清除，这是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 功能活动在不同人群中差异很大 Rev/RRE 活性不是充当开关，而是调节病毒复制。 重新激活后，功能活性较低的 Rev/RRE 对的潜伏感染细胞中的原病毒将被激活。 预计许多病毒蛋白和病毒颗粒的产量会减少，同时继续生产 含有低 Rev/RRE 的原病毒的细胞具有正常水平的免疫调节病毒 Nef 蛋白。 因此，在所谓的“踢杀”治疗策略中，活动可能更难以清除。 本提案中的实验旨在检验这一假设。 建立了潜伏模型和一组具有 Rev/RRE 活性的工程病毒，可以直接 评估用各种潜伏重新激活剂重新激活潜伏感染细胞如何影响病毒激活 Subaim1c 将测量从分离的有能力复制病毒中的 Rev/RRE 功能。 这些实验将确定 Rev/RRE 功能与序列相比如何变化。 来源于初诊患者的外周血，是否经过筛选 具体目标 2 将测量潜伏期重新激活的 CTL 杀灭。 细胞直接确定 Rev/RRE 活性变化如何影响细胞对免疫的敏感性 消除。 更好地了解 Rev-RRE 调节系统在病毒潜伏期如何发挥作用可能是其中之一 这些信息是理解为什么“踢杀”治疗策略迄今为止不成功的关键。 反过来，可以通过调节潜伏病毒的 Rev-RRE 活性来优化“踢”。 因此，这项研究对迄今为止最有希望的治疗艾滋病毒的策略有直接影响，并可以提供关键的信息 知识来推进这一迄今为止难以实现的目标。