Mechanisms of Formation and Persistence of Active HIV Reservoirs

摘要 我们建议解决与实现艾滋病毒感染的功能性治愈有关的几个重要问题 接受抑制性抗逆转录病毒治疗(ART)的个人。具体地说，我们将研究时间和 在原发人类免疫缺陷病毒1型中建立“活性”或感染性储存库的机制 (HIV-1)感染；根据ART确定活跃的艾滋病毒宿主持续多年的机制； 识别拥有活跃储集层的细胞，并识别储集层是否存在于发起ART的个体中 在初次感染期间，持续存在抗原特异的细胞，因此容易与目标细胞一起激活 用于销毁战略的抗原。我们的受试者群体来自一个独特的和长期的队列 在HIV感染的最初几天到几个月内登记的个人，他们之前接受了抑制疗法 从感染之日起至4个月，并在随访期间进行密集的样本采集，最高可达 ~14年。101名受试者在FieBig I-V阶段启动ART，并继续接受ART和 病毒学抑制至少2年，构成主要研究对象。标本来自4个 在慢性HIV感染中开始抗逆转录病毒治疗并在接受抗逆转录病毒治疗时经历病毒反弹的其他受试者 中断也将被用来回答我们的一个问题。我们将对血浆RNA中的病毒进行量化， 外周血单个核细胞(PBMC)中的病毒DNA和AS 2ltR环状未整合病毒DNA，以及 检测前病毒DNA在静息CD4+T细胞中的感染能力。我们还将确定捐款 HIV感染细胞的增殖，以及调控细胞的细胞基因中断之间的关系 周期通过HIV整合到染色体中，使血液中的活性蓄水池持续存在，并在特定情况下 PBMC、细支气管肺泡灌洗液和肠道相关淋巴组织活检中的亚群。 此外，我们将确定我们确定的假定宿主是否允许艾滋病毒凭借其自身的优势保持活跃 细胞内抗逆转录病毒药物水平不足。我们的具体目标是确定：1)机制 和建立活跃的艾滋病毒储存库的动力学；2)(A)增殖的艾滋病毒- 感染细胞，以及(B)低水平的病毒复制，以维持ART期间活跃的水库，以及；3) HIV蓄积细胞中由病毒特异性的CD4+T细胞组成的比例。

ABSTRACT 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. Our 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-level 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.