T-cell depletion and maintenance of the HIV-1 latent reservoir in distinct tissue compartments

T 细胞耗竭和不同组织区室中 HIV-1 潜伏库的维持

• 批准号: 10591589
• 负责人: Melissa Laird Smith
• 金额: $ 71.09万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-04-01 至 2027-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10591589
• 关键词: Allografting; Antigens; Antithymoglobulin; Atlases; Automobile Driving; Binding; Biological Assay; Biological Sciences; Blood; CD4 Positive T Lymphocytes; Cell Separation; Cells; Circulation; Clinical; Clonal Expansion; Clonality; Cytomegalovirus; DNA; Data; Freezing; HIV; HIV-1; Half-Life; Human Herpesvirus 8; Immune; Immunofluorescence Immunologic; Immunologic Markers; Immunologics; Immunosuppression; Immunosuppressive Agents; Individual; Influenza; Interleukin 2 Receptor; Interruption; Kidney; Kidney Transplantation; Lymphoid Cell; Lymphoid Tissue; Maintenance; Measurement; Measures; Metadata; Modeling; Organ; Organ Transplantation; Outcome; Peripheral Blood Mononuclear Cell; Persons; Phenotype; Play; Population; Positioning Attribute; Process; Proliferating; Proviruses; Resolution; Rest; Sampling; Simplexvirus; Site; Solid; Source; Structure of germinal center of lymph node; T cell response; T-Cell Depletion; T-Cell Proliferation; T-Lymphocyte; Time; Tissue Sample; Tissues; Transplant Recipients; Transplantation; Viral; Viral reservoir; Virus; Virus Latency; allograft rejection; antiretroviral therapy; basiliximab; cohort; insight; integration site; interest; kidney allograft; laser capture microdissection; latent HIV reservoir; lymph nodes; migration; multimodality; next generation sequencing; novel; post-transplant; promoter; prophylactic; single molecule

The primary challenge in curing HIV-1 is the persistence of a latent viral reservoir (LVR) in resting CD4+ (rCD4) T cells that harbor stably integrated latent HIV. Examining changes in the LVR composition is incredibly difficult due to the long half-life. Recent data show that clonal expansion of latently infected rCD4 T cells through a combination of antigenic stimulation, homeostatic proliferation, and integration site promotor disruption are major contributors to LVR maintenance. It is unclear which tissue source is the primary driver of LVR maintenance, as well as what level of contribution each of these three potential mechanisms driving proliferation may play in that process. Solid organ transplantation in people living with HIV and the associated different T cell induction strategies prescribed for prophylactic allograft rejection treatment provide a unique opportunity to examine how the LVR rebounds after a large proportion of the T cell repertoire is destroyed. The HOPE in Action HIV+ kidney organ transplantation trial provides access to >120 matched flash frozen lymph nodes (LN), renal allograft tissue, and longitudinally collected peripheral blood mononuclear cells (PBMC) from PLWH. We hypothesize that the LVR is primarily maintained through antigen stimulation of latently infected cells in micro foci within lymph nodes, which subsequently migrate into the circulation and other tissues in the body, thereby reestablishing the LVR post-T cell depletion therapy. Aim 1: Examine long-term LVR dynamics post-renal transplantation and its association with clinical outcomes. We will measure the HIV LVR annually for up to 10 years using the intact proviral DNA assay (IPDA), which distinguishes fully intact HIV from defective, deleted, and hypermutated proviral DNA, in individuals receiving transplant-related immunosuppressive drugs that are of interest to HIV cure strategies. Aim 2: Develop a tissue specific atlas of the LVR in LN, blood, and organ tissue (kidney) pre-transplantation, and examine reseeding of the circulating, LN, and kidney allograft LVR post T cell induction. We will assemble a multi-modal atlas of HIV+ LN by integrating the CODEX multiplexed immunofluorescence (mIF) platform to phenotype lymphoid cells and laser capture microdissection (LCM) and site-directed next-generation sequencing of the proviruses in cells isolated from distinct LN zones. HIV SMRTcap, a novel HIV-specific single molecule sequencing assay will provide simultaneous resolution of proviral sequences and matched integration sites, to evaluate clonality and intactness of latent provirus within the LN, PBMC and kidney. Aim 3: Determine the relative contribution of homeostatic proliferation, antigenic stimulation, and integration site promoter disruption on LVR maintenance and re-establishment post-transplant. These proposed studies will enable us to characterize the longitudinal LVR spatially, genetically, and phenotypically in multiple compartments. This project will provide critical information on feasibility and mechanisms of potential HIV cure strategies by modeling re-seeding of viral populations in kidney allograft and lymphoid tissues and determining driving mechanisms of clonal proliferation.

治愈HIV-1的主要挑战是静止CD4+（RCD4）中潜在病毒储存库（LVR）的持久性 具有稳定整合潜在艾滋病毒的T细胞。检查LVR组成的变化非常困难 由于半衰期。最近的数据表明，通过A的克隆膨胀通过A 抗原刺激，稳态增殖和整合部位启动子的混合是 LVR维护的主要贡献者。目前尚不清楚哪个组织源是LVR的主要驱动力 维护，以及这三种潜在机制的贡献水平，驱动增殖 可以在这个过程中播放。艾滋病毒和相关的不同T的固体器官移植 针对预防性同种异体移植排斥治疗规定的细胞诱导策略为您提供了独特的机会 检查大部分T细胞库被破坏后LVR如何反弹。希望 动作HIV+肾脏器官移植试验可访问> 120个匹配的闪光冷冻淋巴结（LN）， 肾脏同种异体移植组织和纵向收集的外周血单核细胞（PBMC）。我们 假设LVR主要是通过抗原刺激在微观中维持的。 淋巴结中的焦点，随后迁移到体内的循环和其他组织中，从而 重新建立LVR T细胞耗竭疗法。 目标1：检查长期LVR动力学后肾脏移植及其与临床结局的关联。 我们将使用完整的前病毒DNA分析（IPDA）每年测量每年的HIV LVR长达10年，该测定法（IPDA） 在接受的个体中，将完全完整的HIV与有缺陷，删除和超重的病毒DNA区分开 艾滋病毒治愈策略感兴趣的与移植相关的免疫抑制药物。目标2：开发组织 LN，血液和器官组织（肾脏）预先进行的LVR特定地图集 T细胞诱导后循环，LN和肾脏同种异体移植LVR。我们将组装HIV+的多模式地图集 LN通过将法典多重免疫荧光（MIF）平台整合到表型淋巴样细胞和 激光捕获显微解剖（LCM）和位置定向的细胞病毒的下一代测序 从不同的LN区域分离出来。 HIV SMRTCAP，一种新型的HIV特异性单分子测序测定法 提供前病毒序列和匹配的集成位点的同时解决，以评估克隆性和 LN，PBMC和肾脏内潜在病毒的完整性。目标3：确定相对贡献 LVR维护上的稳态增殖，抗原刺激和集成站点启动子破坏 并重建移植后。 这些提出的研究将使我们能够在空间，遗传和 在多个隔间中表型。该项目将提供有关可行性和 通过对肾脏同种异体移植中病毒种群的重新种植进行建模，潜在的HIV治疗策略的机制 淋巴组织和确定克隆增殖的驱动机制。