Mechanisms of Formation and Persistence of Active HIV Reservoirs

描述(由申请人提供)：我们建议通过抑制性抗逆转录病毒疗法(ART)解决与实现个体艾滋病毒感染的功能性治愈相关的几个重要问题。具体地说，我们将研究在原发人类免疫缺陷病毒1型(HIV-1)感染中建立“活跃”或感染性宿主的时机和机制；确定艾滋病毒活跃宿主在抗逆转录病毒治疗下持续多年的机制，确定拥有活跃宿主的细胞；以及辨别在初次感染期间启动抗逆转录病毒治疗的个体的宿主是否持续存在抗原特异性细胞，从而能够被其目标抗原激活以用于破坏策略。我们的受试者群体来自一个独特和长期的队列，这些人在艾滋病毒感染的最初几天到几个月期间登记，他们在感染之日起4个月前接受抑制疗法，并在长达~14年的随访期间进行了密集的样本收集。101名在费比格I-V阶段开始抗逆转录病毒治疗，并继续接受抗逆转录病毒治疗至少2年的受试者，构成了主要的研究对象组。另外4名在慢性HIV感染中开始抗逆转录病毒治疗并在抗逆转录病毒治疗中断时经历病毒反弹的受试者的样本也将被用来回答我们的一个问题。我们将对血浆RNA中的病毒、外周血单个核细胞(PBMC)中的总病毒DNA和循环中未整合的病毒DNA进行定量，并检测前病毒DNA在静止的CD4T细胞中的感染能力。我们还将确定艾滋病毒感染细胞增殖的贡献，以及调节细胞周期的细胞基因中断之间的关系 通过艾滋病毒整合到染色体、血液和PBMC中特定亚群中活性储存库的持续存在、细支气管肺泡灌洗液和肠道相关淋巴组织的活组织检查。此外，我们将确定我们确定的假定宿主是否允许艾滋病毒由于细胞内抗逆转录病毒药物水平不足而保持活跃。我们的具体目标是确定：1)建立活跃的艾滋病毒储存库的机制和动力学；2)(A)艾滋病毒感染细胞的增殖，(B)低水平的病毒复制对抗逆转录病毒治疗期间活跃储存库的持续的相对贡献；以及；3)艾滋病毒的比例 蓄积细胞由病毒特异性的CD4T细胞组成。

DESCRIPTION (provided by applicant): We propose to address several important questions relevant to achieving a functional cure of HIV infection in individuals on suppressive antiretroviral therapy (ART). Specifically, we will examine the timing and mechanisms that establish the "active" or infectious reservoirs in primary human immunodeficiency virus type 1 (HIV-1) infection; identify the mechanisms by which active HIV reservoirs persist for many years under ART, identify the cells that harbor active reservoirs, and; discern whether reservoirs in individuals initiating ART during primary infection persist in antigen-specific cells and are thus amenable to activation with their target antigens for destruction strategies. Our subject population derives from a unique and long standing cohort of individuals enrolled while in the first few days to months of HIV infection, who went onto suppressive ART prior to 4 months from the date of infection and have had intensive specimen collection during follow-up for up to ~14 years. One hundred and one subjects who initiated ART in Fiebig stages I-V, and remained on ART and virologically suppressed for at least 2 years, form the primary set of subjects for study. Specimens from 4 additional subjects who started ART in chronic HIV infection and experienced a viral rebound when ART was interrupted will be also be used to address one of our questions. We will quantify virus in plasma RNA, as total viral DNA and as 2LTR circular unintegrated viral DNA in peripheral blood mononuclear cells (PBMC), and examine the infectious capacity of proviral DNA in resting CD4+T cells. We will also determine the contribution of proliferating HIV-infected cells, and the relationship between disruption of cellular genes regulating the cell cycle by HIV integration into the chromosome, to the persistence of active reservoirs in blood and in specific subpopulations in PBMC, bronchioalveolar lavage fluids and biopsies of gut-associated lymphoid tissues. Additionally, we will determine whether the putative reservoirs we identify allow HIV to remain active by virtue of inadequate intracellular levels of antiretroviral drugs. Ou specific aims are to determine: 1) The mechanisms and kinetics of establishment of active HIV reservoirs; 2) the relative contributions of (A) proliferating HIV- infected cells, and (B) low-levl viral replication to the persistence of active reservoirs during ART, and; 3) the proportion of HIV reservoir cells composed of virus-specific CD4+T cells.