Dynamics, immune responses, and transcriptomics of the HIV-expressing reservoir on ART

项目摘要/摘要(项目1) 为了开发新的治疗方法，旨在治愈艾滋病毒或减少抗逆转录病毒治疗感染的后遗症，有 迫切需要对艾滋病毒感染细胞的持久性进行更多研究，这有助于免疫激活 在艺术上，并允许病毒学在艺术中断后反弹。而反弹能力较强的储油层往往 假定与潜伏的和/或完整的储存库相同，来自反弹病毒的序列通常是这样的 不匹配来自潜伏储存库的那些，但有时与ART之前的细胞相关HIV RNA匹配 中断。这些发现表明，有必要更好地了解体内表达艾滋病毒的宿主，这些宿主 准备好在ART中断时启动反弹，并可能有助于免疫激活，器官 损害，并减少了ART的预期寿命。然而，先前对“转录活性储存库”的研究 还不能完全描述这些细胞的异质性，这些细胞在艾滋病毒类型方面有所不同 转录的RNA(过程的或完全的，来自有缺陷的或完整的前病毒)以及它是否被翻译成 HIV蛋白质。我们假设，表达不同类型HIV RNA和/或蛋白质的细胞亚群将有所不同 在频率、存活率/清除率、对免疫激活的贡献、细胞基因表达、 和组织分布。为了研究这些假设，这个项目将应用一系列新的和尖端的 对不同临床表型的纵向样本进行分析，以：1)衡量不同的临床表现 (完整/缺陷)，阻止HIV转录，缺陷或完整的HIV转录本，以及HIV Gag蛋白变化 随着时间的推移，血液中的艺术前和艺术后，以及精英控制者和发起 急性或慢性感染期间的抗逆转录病毒治疗；2)确定每种类型的艾滋病毒RNA和蛋白质水平如何相互关联 具有HIV特异性T细胞和其他免疫反应，以及免疫激活/炎症的标志；以及3) 确定宿主细胞基因(如抗病毒因子)的差异表达如何与这些基因的能力相关 细胞表达艾滋病毒并在血液和淋巴结内存活。后续项目将调查不同之处 前病毒和HIV表达细胞的类型在其频率、表型和病毒/细胞基因上有所不同 在体内所有组织中的表达(项目2)，以及它们在 对针对和杀死HIV表达细胞的新疗法的反弹和敏感性的贡献 (项目3)。

Project Summary/Abstract (Project #1) To develop new therapies aimed at HIV cure or reducing the sequelae of ART-treated infection, there is a critical need for more research on the persistence of HIV-infected cells that can contribute to immune activation on ART and allow virologic rebound after ART interruption. While the rebound competent reservoir is often assumed to be the same as the latent and/or the intact reservoir, sequences from the rebound virus usually do not match those from the latent reservoir but sometimes match those from cell-associated HIV RNA prior to ART interruption. These findings suggest the need to better understand the reservoirs that express HIV in vivo, which are poised to initiate rebound on interruption of ART and likely contribute to the immune activation, organ damage, and reduced life expectancy on ART. However, prior studies of the “transcriptionally active reservoir” have not been able to fully characterize the heterogeneity of these cells, which vary in terms of the types of HIV RNA transcribed (processive or complete, from defective or intact proviruses) and whether it is translated into HIV protein. We hypothesize that subsets of cells expressing different types of HIV RNA and/or protein will differ in terms of their frequency, survival/clearance rate, contribution to immune activation, cellular gene expression, and tissue distribution. To investigate these hypotheses, this project will apply a series of new and cutting-edge assays to longitudinal samples fromdifferent clinical phenotypes in order to: 1) measure how different proviruses (intact/defective), blocks to HIV transcription, defective or intact HIV transcripts, and HIV Gag protein change over time pre- and post-ART in the blood and how they differ between elite controllers and individuals who initiate ART during acute or chronic infection; 2) determine how levels of each type of HIV RNA and protein correlate with HIV-specific T and other immune responses as well as markers of immune activation/inflammation; and 3) determine how differential expression of host cell genes (such as antiviral factors) relates to the ability of these cells to express HIV and survive in blood and lymph nodes. Subsequent projects will investigate how different types of proviruses and HIV-expressing cells differ in their frequencies, phenotypes, and viral/cellular gene expression throughout the full spectrum of tissues in the body (Project #2), and how they differ in their contribution to rebound and susceptibility to novel therapies designed to target and kill HIV-expressing cells (Project #3).