Mechanisms of Formation and Persistence of Active HIV Reservoirs

活跃HIV病毒库的形成和持续机制

• 批准号: 9229518
• 负责人: JAMES Ivan MULLINS
• 金额: $ 85.28万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-03-15 至 2019-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9229518
• 关键词: Acute; Address; Adult; Anti-Retroviral Agents; Antigen Targeting; Antigenic Specificity; Antigens; Apoptosis; Back; Biopsy; Blood; Bronchoalveolar Lavage; CD4 Positive T Lymphocytes; Cell Cycle; Cell Proliferation; Cells; Chromosomes; Chronic; Combined Vaccines; Cytomegalovirus; DNA; Data; Early treatment; Enrollment; Event; Genes; Goals; Gut associated lymphoid tissue; HIV; HIV Infections; HIV therapy; HIV vaccine; HIV-1; Human Herpesvirus 4; Immunity; Individual; Infant; Infection; Interruption; Irrigation; Kinetics; Knowledge; Lead; Link; Liquid substance; Methods; Myelogenous; Patient Care; Peripheral Blood Mononuclear Cell; Pharmaceutical Preparations; Plasma; Population; Primary Infection; Prior Therapy; Production; Proliferating; Proteins; Provirus Integration; Proviruses; Public Health; RNA; Reporting; Research; Rest; Simplexvirus; Site; Specimen; Study Subject; T memory cell; Targeted Toxins; Tetanus; Time; Tissues; Viral; Viral reservoir; Virion; Virus; Virus Replication; alpha Toxin; antiretroviral therapy; co-infection; cohort; experience; follow-up; improved; integration site; pandemic disease; prevent; public health relevance; sample collection; site-specific integration; treatment group; viral DNA; viral RNA; viral rebound; virology

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.

描述（由申请人提供）：我们建议解决几个重要的问题，相关实现抑制性抗逆转录病毒治疗（ART）的个人HIV感染的功能性治愈。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”或感染性储库的时间和机制;确定在ART下活性HIV储库持续多年的机制，确定携带活性储库的细胞;辨别在初次感染期间启动ART的个体中的储库是否在抗原中持续存在-特异性细胞，并因此适合于用其靶抗原活化以用于破坏策略。我们的受试者人群来自于一个独特的长期队列，该队列是在HIV感染的最初几天至几个月内入组的个体，他们在感染之日起4个月前接受了抑制性ART，并在随访期间进行了长达14年的密集标本采集。在Fiebig I-V期开始ART治疗并保持ART治疗和病毒学抑制至少2年的101例受试者形成了研究的主要受试者集。另外4例因慢性HIV感染开始ART治疗并在ART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。我们将定量血浆RNA中的病毒、外周血单核细胞（PBMC）中的总病毒DNA和2LTR环状未整合病毒DNA，并检查静息CD 4 +T细胞中前病毒DNA的感染能力。我们还将确定HIV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HIV整合到染色体中，到血液中和PBMC、支气管肺泡灌洗液和肠道相关淋巴组织活检中的特定亚群中活性储库的持续存在。此外，我们将确定我们确定的假定水库是否允许艾滋病毒保持活性，由于抗逆转录病毒药物的细胞内水平不足。我们的具体目标是确定：1）建立活性HIV储库的机制和动力学; 2）ART期间（A）增殖的HIV感染细胞和（B）低水平病毒复制对活性储库持续存在的相对贡献;以及3）HIV感染细胞和HIV感染细胞的比例。 由病毒特异性CD 4 +T细胞组成的储库细胞。