Investigation of persistent HIV immune stimulation in lymphoid tissues during therapy as a cause of sustained immune activation

研究治疗期间淋巴组织中持续的 HIV 免疫刺激作为持续免疫激活的原因

• 批准号: 10598469
• 负责人: Timothy W Schacker
• 金额: $ 74.15万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-03 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10598469
• 关键词: Address; Age; Anatomy; Anti-Retroviral Agents; Antigens; B-Lymphocytes; Biological Assay; C-reactive protein; CD4 Positive T Lymphocytes; Cells; Circulation; Coagulation Process; Collection; Cytomegalovirus; Data; Detection; Development; Fibrin fragment D; Foundations; Frequencies; Future; Gene Expression Profile; Genes; Genetic Transcription; HIV; HIV Infections; HIV Seronegativity; HIV envelope protein; Health; Helper-Inducer T-Lymphocyte; Herpesviridae; Herpesviridae Infections; Human Herpesvirus 4; Image Analysis; Immune; Immune response; Immunologic Stimulation; In Situ Hybridization; Individual; Infection; Interleukin-6; Interruption; Interstitial Nephritis; Investigation; Link; Lymphoid Tissue; Measurement; Measures; Metabolic; Methods; Mononuclear; Morbidity - disease rate; Outcome; Pathology; Patients; Pharmaceutical Preparations; Phenotype; Plasma; Production; Proliferating; Pulmonary Hypertension; RNA; Research; Rest; Serum Markers; Simplexvirus; Site; Source; Strategic Planning; T-Lymphocyte; T-Lymphocyte Subsets; TNF gene; Technology; Testing; Tissues; United States National Institutes of Health; Viral; Viral Antigens; Viral Load result; Viral reservoir; Virion; Virus; Virus Replication; Work; antibody test; antiretroviral therapy; cardiovascular disorder risk; cell envelope; comorbidity; detection limit; endothelial dysfunction; experience; genetic signature; high throughput screening; immune activation; immune reconstitution; improved; microbial; mortality; neutralizing antibody; quantitative imaging; transcriptomics; translational approach; viral detection

Antiretroviral therapy (ART) sufficiently suppresses HIV replication to reduce plasma viral load (pVL) below the limit of detection, but immune activation (IA) is not normalized and the elevated levels of IA markers- IL-6, TNF, TGFß, C reactive protein (CRP), and D-Dimer remain elevated are associated with increased risk for cardiovascular disease, endothelial disfunction and clotting abnormalities, pulmonary hypertension, interstitial nephritis, development of non-AIDS associated malignancies, and CNS abnormalities. Multiple mechanisms have been proposed to explain the persistence of IA under ART including microbial translocation and herpes virus infections (e.g., HSV, CMV, EBV). However, here we propose that HIV itself is a major cause of persistent IA because of ongoing virus production in lymphoid tissues (LT) while on ART. In this revised proposal, we have one specific aim that tests two hypotheses: 1) Sustained IA during ART is driven by production of virions and/or expression of viral antigens in reactivated latently infected cells with or without persistent low-level virus replication; and 2) that persistent low-level virus replication during ART is the result of intracellular concentrations (IC) of antiretroviral (ARV) drugs in LT that do not completely inhibit replication and virus production. For our first hypothesis we will seek direct evidence of correlations between persistent IA and virus production/antigen expression in LT by identification, at the single cell level, of: 1) virus-producing CD4 T cells lacking markers of activation and proliferation that we have previously shown can sustain low levels of virus production; 2) T follicular helper cells (Tfh) in B cell follicles that have recently been shown to be an independent reservoir for viral persistence; and 3) reactivated latently infected T cells producing virus or p24. We propose investigations of these drivers of IA by our validated and highly sensitive in situ hybridization (ISH) methods to detect, phenotype and locate virus (v) RNA+ and virus-producing cells in LT; by a validated highly sensitive high throughput “envelope detection by induced transcription-based sequencing” (EDITS) assay; and a broadly neutralizing antibody (bNab) method to enrich for HIV-envelope (ENV)+ cells. For the hypothesized correlations of the virus drivers with IA, we will measure IA with standard flow-based antibody assays and with single cell transcriptomic analysis of LT mononuclear cells to identify unique gene signatures associated with IA. Our second hypothesis will be tested by quantification of ARV-intracellular concentrations (ICs) in LT and determine the relationships among ARV-ICs and the frequency of detecting the three putative virus drivers of IA. Establishing that HIV itself is a cause of IA would point future developments to fully suppress virus production in the LT reservoir with benefits both in reducing IA and associated pathologies and as an essential component of HIV Cure Strategies.

抗逆转录病毒疗法（ART）充分抑制了HIV复制，以降低血浆病毒载量（PVL） 检测极限，但免疫激活（IA）未归一化，IA标记的水平升高-IL-6，TNF， TGFß，C反应性蛋白（CRP）和D-二聚体保持升高与增加的风险有关 心血管疾病，内皮疾病和凝结异常，肺动脉高压，间隙 肾炎，非AID相关的恶性肿瘤的发展和中枢神经系统异常。多种机制 已经提出要解释IA在艺术下的持久性，包括微生物易位和疱疹 病毒感染（例如HSV，CMV，EBV）。但是，在这里我们建议艾滋病毒本身是 持续性IA是因为淋巴组织中持续产生的病毒（LT）。在此修订中 提案，我们有一个特定的目的，它检验了两个假设：1）在艺术期间持续的IA驱动。 在有或没有的，有或没有 持续的低级病毒复制； 2）在艺术期间持续的低级病毒复制是 LT中抗逆转录病毒（ARV）药物的细胞内浓度（IC）不会完全抑制复制和 病毒生产。对于我们的第一个假设，我们将寻求直接证据证明持续的IA与 通过在LT中的病毒产生/抗原表达，在单细胞水平上鉴定为：1）产生CD4 T的病毒 我们以前已经显示的缺乏激活和增殖标记的细胞可以维持较低的水平 病毒生产； 2）B细胞中的T卵泡辅助细胞（TFH）最近被证明是一种 独立的病毒持久性水库； 3）重新激活了产生病毒或p24的潜在感染的T细胞。 我们通过我们经过验证且高度敏感的原位杂交（ISH）对这些IA的这些驱动因素进行研究。 检测，表型和定位病毒（V）RNA+和产生病毒细胞的方法；通过经过验证的高度 敏感的高吞吐量“通过诱导转录测序检测包膜检测”（编辑）测定；和 一种广泛的中和抗体（BNAB）方法，可富集HIV-Envelope（Env）+细胞。对于假设的 病毒驱动因素与IA的相关性，我们将使用标准的基于流动的抗体测定和与 LT单核细胞的单细胞转录组分析，以鉴定与 ia。我们的第二个假设将通过量化LT中的ARV-细胞浓度（IC）来检验 确定ARV-ICS之间的关系以及检测三种假定病毒驱动因素的频率 ia。确定艾滋病毒本身是造成IA的原因将指出未来的发展以完全抑制病毒 在减少IA和相关病理方面的利益中，在LT储层中生产有益，也是必不可少的 HIV治疗策略的组成部分。