Mechanisms of Formation and Persistence of Active HIV Reservoirs

描述（由申请人提供）：我们建议解决几个重要的问题，这些问题与在抑制性抗逆转录病毒治疗（ART）中实现HIV感染的功能性治愈有关。具体来说，我们将研究在原发性人类免疫缺陷病毒1型（HIV-1）感染中建立“活性”或感染性宿主的时间和机制；确定在抗逆转录病毒治疗下活性HIV病毒库持续多年的机制，确定携带活性HIV病毒库的细胞；辨别在原发性感染期间启动抗逆转录病毒治疗的个体体内的储库是否在抗原特异性细胞中持续存在，从而能够被其靶抗原激活以实施破坏策略。我们的受试者群体来自一个独特的长期队列，这些个体在感染艾滋病毒的最初几天到几个月内被招募，在感染之日起4个月之前接受了抑制性抗逆转录病毒治疗，并在长达14年的随访期间进行了密集的标本收集。101名在Fiebig I-V期开始接受抗逆转录病毒治疗，并持续接受抗逆转录病毒治疗和病毒学抑制至少2年的受试者构成了研究的主要对象组。另外4名在慢性艾滋病毒感染时开始抗逆转录病毒治疗并在抗逆转录病毒治疗中断时出现病毒反弹的受试者的标本也将用于解决我们的一个问题。我们将量化血浆RNA中的病毒，总病毒DNA和外周血单核细胞（PBMC）中的2LTR环状未整合病毒DNA，并检测静息CD4+T细胞中的前病毒DNA的感染能力。我们还将确定增殖艾滋病毒感染细胞的贡献，以及调节细胞周期的细胞基因破坏之间的关系

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.