Roles of endothelial cells in HIV infection and latency in resting CD4+ T cells

描述（由申请人提供）：在许多HIV-1感染患者中，高效抗逆转录病毒疗法（HAART）成功抑制了病毒载量并恢复了CD4 + T细胞数量。然而，在静息CD4 + T细胞中鉴定的主要潜伏性储库对根除构成了很大的障碍，并确保了病毒在患者中的持续存在。对这种潜在油气藏是如何形成的认识相当有限。更全面地了解有助于建立水库的机制将影响对抗病毒持久性的策略。 最近在体内或离体进行的一些研究表明，淋巴组织的微环境和体内细胞-细胞相互作用在静息CD4 + T细胞的潜伏期形成中起重要作用。此外，内皮细胞，在体内的淋巴组织中的T细胞在生理上很容易相互作用，诱导生产性感染的HIV在休息的CD4 + T细胞，并可能在这些细胞的潜伏期形成中发挥重要作用。 本研究的总体目标是研究内皮细胞对HIV-1感染和静息CD4 + T细胞潜伏期的作用。具体目的是1）使用新开发的体外长寿命原代T细胞系统研究内皮细胞对静息CD4 + T细胞中HIV潜伏期形成的影响; 2）使用患者样品和体外产生的潜伏感染的静息CD4 + T细胞检查内皮细胞对潜伏HIV的再活化;将来自患者或来自体外产生的潜伏感染的静息CD4 + T细胞与内皮细胞共培养以检查前病毒的再活化。 3)使用蛋白质印迹法鉴定可能参与内皮细胞刺激的静息T细胞中HIV感染的细胞因子。 从这项研究中获得的知识将显着提高对艾滋病毒潜伏储库形成的理解，并将影响对抗病毒持久性的策略。它还将深入了解有助于HIV感染CD4 + T细胞的机制，并可能有助于潜在的创新干预。 公共卫生相关性：根除HIV-1疾病的最大障碍是静息CD4 + T细胞中的潜伏病毒库。这项研究旨在确定内皮细胞在HIV-1感染和静息CD4 + T细胞潜伏期中的作用。对这个问题的回答将增加我们对艾滋病毒感染和持续性的理解，并有助于治疗艾滋病毒疾病的潜在创新干预。

DESCRIPTION (provided by applicant): In many patients with HIV-1 infection, highly active antiretroviral therapy (HAART) successfully suppresses viral loads and restores CD4+ T cell numbers. However, a major latent reservoir identified in resting CD4+ T cells poses a great barrier to eradication and ensures viral persistence in patients. The understanding of how such a latent reservoir is formed is quite limited. A more complete understanding of the mechanisms contributing to the establishment of the reservoir will influence the strategies in battling viral persistence. Some recent studies done in vivo or ex vivo suggested that the microenvironment of the lymphoid tissue and cell-cell interactions in vivo played important roles in latency formation in resting CD4+ T cells. Moreover, endothelial cells, which physiologically interact readily with T cells in the lymphoid tissues in vivo, were shown to induce productive infection of HIV in resting CD4+ T cells and might play a significant role in latency formation in these cells. The overall goal of this study is to investigate the roles of endothelial cells on HIV-1 infection and latency in resting CD4+ T cells. The specific aims are 1) to investigate the effect of endothelial cells on HIV latency formation in resting CD4+ T cells using a newly developed in vitro long-lived primary T cell system; 2) to examine reactivation of latent HIV by endothelial cells using patient samples and in- vitro-generated latently infected resting CD4+ T cells; latently infected resting CD4+ T cells from patients or from in vitro generation will be co-cultured with endothelial cells to examine reactivation of proviruses. 3) to identify cellular factors potentially involved in allowing HIV infection in endothelial-cell- stimulated resting T cells using western blots. The knowledge gained from this study will significantly improve the understanding of HIV latent reservoir formation and will influence the strategies in battling viral persistence. It will also provide insight into the mechanisms contributing to HIV infection of CD4+ T cells and may contribute to potential innovative intervention. PUBLIC HEALTH RELEVANCE: The biggest barrier to eradication of HIV-1 diseases is the latent viral reservoir in resting CD4+ T cells. The proposed study aims to determine the roles endothelial cells play on HIV-1 infection and latency in resting CD4+ T cells. An answer to this question will increase our understanding of HIV infection and persistence and contribute to potential innovative intervention in treating HIV diseases.