An Ex Vivo Model of HIV-1 Latency and Reactivation using Primary Memory Cells

描述（由申请方提供）：拟定研究使用了一种新的病毒潜伏期和再激活实验模型，该模型利用原代记忆细胞作为靶点。使用这个系统，我们计划详细了解HIV-1用于建立体内潜伏期的机制，以及触发病毒再激活的生理信号。来自这些领域的知识将适用于未来的转化研究，这些研究将寻求模仿或拮抗相关信号通路的化合物，最终目标是破坏潜在的水库。 公共卫生相关性：HIV-1在感染者体内建立了一个相对较小但寿命较长的潜伏感染细胞库。潜伏性感染与低或无病毒基因表达相关，因此似乎是非细胞病变性的。潜伏的病毒可以重新激活，引起新的具有完全致病潜力的生产性感染。这种长期潜伏的病毒库的细胞类型被认为是由静止的记忆T细胞组成的。我们提出了一种新的离体模型的研究艾滋病毒的潜伏期，以及它的再激活。该模型忠实地概括了体内潜伏库的显著特征。例如，该系统中的潜伏前病毒缺乏任何可检测的基因表达，然而，当被许多外部刺激重新激活时，它们完全有能力进行基因表达。该模型使用原代人CD 4+淋巴细胞，病毒整合是多克隆的。这个强大而直接的实验系统将使我们能够（i）评估各种CD 4 + T细胞亚群对潜伏性前病毒的相对允许性;（ii）探索可能阻止潜伏性感染建立和/或诱导潜伏性前病毒再活化的信号通路;和（iii）探索使用相关信号通路的选择激动剂和拮抗剂来实验性地操纵潜伏期和再激活。 最终，根除HIV-1需要对控制进入和退出潜伏期的生物过程有深刻的理解。

DESCRIPTION (provided by applicant): The proposed studies use a novel experimental model of viral latency and reactivation that utilizes primary memory cells as targets. Using this system, we plan to understand in detail the mechanisms that HIV-1 uses to establish latency in vivo, as well as the physiological signals that trigger viral reactivation. Knowledge from these areas will be applicable in future translational studies that will seek compounds mimicking or antagonizing the relevant signaling pathways, with the ultimate goal of destroying the latent reservoir. PUBLIC HEALTH RELEVANCE: HIV-1 in infected individuals establishes a relatively small, but long-lived reservoir of latently infected cells. Latent infection is associated with low or no viral gene expression and, accordingly, appears to be non-cytopathic. Latent viruses can undergo reactivation, giving rise to new productive infections that carry full pathogenic potential. The cell type that harbors this long-lived, latent viral reservoir is thought to consist of quiescent memory T cells. We present a novel ex-vivo model for the study HIV latency as well as its reactivation. This model faithfully recapitulates salient features of the in vivo latent reservoir. For example, latent proviruses in this system are devoid of any detectable gene expression, yet, they are perfectly competent for gene expression when reactivated by a number of external stimuli. This model uses primary, human CD4+ lymphocytes and viral integration is polyclonal. This powerful, yet straighforward experimental system will allow us to (i) assess the relative permissivenes of various CD4+ T cell subsets to harbor latent proviruses; (ii) explore signaling pathways that may preclude the establishment of latent infections and/or induce reactivation of latent proviruses; and (iii) explore the use of select agonists and antagonists of the relevant signaling pathways to experimentally manipulate latency and reactivation. Ultimately, HIV-1 eradication will require a profound understanding of the biological processes that control entry and exit from latency.