Role of Viral Reservoirs in the Pathogenesis of HIV Disease

病毒库在 HIV 疾病发病机制中的作用

• 批准号: 7732546
• 负责人: Anthony S. Fauci
• 金额: $ 108.96万
• 依托单位: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7732546
• 关键词: Anti-HIV Agents; Antiviral Therapy; Biological Assay; Blood; CD4 Positive T Lymphocytes; Cell Count; Cells; Chronic Phase; Class; Coculture Techniques; Cross Infection; Cross-Sectional Studies; DNA; Data; Disease; Frequencies; Goals; Gut associated lymphoid tissue; HIV; HIV Infections; Individual; Infection; Investigation; Lead; Measurable; Numbers; Pathogenesis; Phylogenetic Analysis; Research; Residual state; Rest; Role; Staging; Time; Tissues; Viral; Virus; antiretroviral therapy; in vivo; peripheral blood; Anti-HIV Agents; Antiviral Therapy; Biological Assay; Blood; CD4 Positive T Lymphocytes; Cell Count; Cells; Chronic Phase; Class; Coculture Techniques; Cross Infection; Cross-Sectional Studies; DNA; Data; Disease; Frequencies; Goals; Gut associated lymphoid tissue; HIV; HIV Infections; Individual; Infection; Investigation; Lead; Measurable; Numbers; Pathogenesis; Phylogenetic Analysis; Research; Residual state; Rest; Role; Staging; Time; Tissues; Viral; Virus; antiretroviral therapy; in vivo; peripheral blood

For the past several years, we have been investigating the role of latently infected, resting CD4+ T cells and persistent viral replication in the pathogenesis of HIV disease and the impact of this reservoir on the treatment of HIV-infected individuals. We previously demonstrated that the latent viral reservoir in the resting CD4+ T cell compartment persists in virtually all infected individuals receiving effective antiviral therapy. Consequently, this viral reservoir is a major impediment to the eradication of HIV in vivo. In addition, we realized that HIV continually replicates at low levels in chronically infected individuals who receive effective antiviral therapy that renders them consistently aviremic for prolonged periods of time. Over the past year, we have focused our research on: 1) delineation of the mechanism by which HIV persists in infected individuals receiving effective antiviral therapy for extended periods of time and 2) investigation of the dynamics of decay of viral reservoirs in infected individuals who initiate antiretroviral therapy at different stages of disease. First, we investigated the presence and status of residual HIV in individuals who had received effective antiretroviral therapy for prolonged periods of time and examined the underlying mechanisms by which HIV persists in CD4+ T cells of such individuals. We demonstrated that CD4+ T cells in the gut-associate lymphoid tissue (GALT) carried the highest level of HIV proviral DNA compared to CD4+ T cells in the blood of infected individuals receiving effective antiretroviral therapy for prolonged periods of time. Phylogenetic analyses of HIV env sequences revealed active cross-infection between blood- and GALT-associated CD4+ T cells. Furthermore, our data also suggest that intensification of antiretroviral therapy, especially through the addition of newer classes of anti-HIV drugs, will be necessary to contain low levels of on-going viral replication in the tissue compartment of chronically infected individuals in order to achieve complete clearance of virus in vivo. Second, we investigated the dynamics of decay of viral reservoirs in infected individuals who initiated antiretroviral therapy at different stages of disease. In a cross-sectional study, we demonstrated that the median copy number of HIV proviral DNA in subjects who initiated antiretroviral therapy within 6 months of infection was significantly lower compared to subjects initiating antiretroviral therapy during the chronic phase of infection (p=0.003). In order to examine the frequency of CD4+ T cells carrying infectious virus, a High Input Co-culture assay, which allows examination of large numbers of cells, was conducted using highly enriched CD4+ T cells from 8 infected individuals in whom no measurable HIV proviral DNA had been detected in their cells. The frequency of cells carrying infectious virus in HIV-infected individuals who initiated therapy within 6 months of infection was significantly lower compared to HIV-infected individuals who initiated therapy during the chronic phase of infection (p=0.03). Remarkably, no infectious virus could be recovered from the peripheral blood or GALT CD4+ T cells of one infected individual (<1 infected cell per 0.8x109 CD4+ T cells) in whom antiretroviral therapy was initiated early in infection. Our data suggest that the combination of initiation of antiretroviral therapy early in the course of HIV infection and prolonged suppression of viral replication can result in a profound diminution of HIV reservoirs that may lead to clearance of virus in infected individuals.

在过去的几年中，我们一直在研究静止感染的CD4+ T细胞的作用，并在HIV疾病发病机理中持续的病毒复制以及该储层对HIV感染个体的治疗的影响。我们先前证明，在几乎所有接受有效抗病毒药疗法的感染者中，静止的CD4+ T细胞室中的潜在病毒储存剂持续存在。因此，这种病毒储层是消除体内HIV的主要障碍。此外，我们意识到，在接受有效的抗病毒疗法的慢性感染者中，HIV持续以低水平的速度复制，这使他们在长时间的时间内始终如一。在过去的一年中，我们将研究重点放在：1）在长时间接受有效抗病疗法的感染者中持续艾滋病毒持续存在的机制和2）研究病毒储存剂的衰变动力学的机制，这些机制在不同阶段启动抗病毒疗法的受感染个体的病毒储量衰变动态。 首先，我们研究了在长时间接受有效的抗逆转录病毒疗法的个体中残留HIV的存在和状态，并检查了此类个体CD4+ T细胞中HIV持续存在的基本机制。我们证明了肠道酶相关淋巴组织（GALT）中的CD4+ T细胞具有最高水平的HIV病毒前DNA水平，与受感染的个体的血液中的CD4+ T细胞相比，接受有效的抗逆转录病毒疗法长时间。 HIV ENV序列的系统发育分析揭示了血液和GALT相关的CD4+ T细胞之间的主动交叉感染。此外，我们的数据还表明，抗逆转录病毒疗法的强化，尤其是通过添加了较新的抗HIV药物，必须在慢性感染个体的组织隔室中含有低水平的持续病毒复制，以便在体内实现病毒的完全清除。 其次，我们研究了在疾病不同阶段开始抗逆转录病毒疗法的受感染个体中病毒储存剂衰减的动力学。在一项横断面研究中，我们证明，与在感染期间开始抗逆转录病毒疗法的受试者相比，在感染后6个月内启动抗逆转录病毒疗法的受试者的HIV前病毒DNA的中间拷贝数显着降低（P = 0.003）。为了检查携带感染性病毒的CD4+ T细胞的频率，使用高度富集的CD4+ T细胞进行了高输入共培养测定，该测定允许检查大量细胞，这些CD4+ T细胞在其细胞中未检测到未检测到的可测量的HIV HIV病毒病毒。与在慢性感染期间开始治疗的HIV感染个体相比，在感染后6个月内开始治疗的HIV感染个体中携带感染病毒的细胞频率显着降低（P = 0.03）。值得注意的是，没有一个感染个体的外​​周血或GALT CD4+ T细胞（每0.8x109 CD4+ T细胞）从外周血CD4+ T细胞中回收传染性病毒，其中抗逆转录病毒疗法在感染的早期就开始了。我们的数据表明，在HIV感染过程中早期抗逆转录病毒疗法的起始和长期抑制病毒复制可能会导致HIV储层的大幅减少，这可能导致感染个体中病毒的清除。