CD8 T cell suppression of HIV latency establishment and maintenance in virally suppressed individuals

CD8 T 细胞抑制病毒抑制个体中 HIV 潜伏期的建立和维持

• 批准号: 10242771
• 负责人: Deanna A Kulpa
• 金额: $ 39万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-24 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10242771
• 关键词: Address; Animals; Antiviral Agents; CD4 Positive T Lymphocytes; CD8-Positive T-Lymphocytes; CD8B1 gene; Cells; Clinical; Clinical Research; Cytotoxic T-Lymphocytes; DNA Modification Process; Disease; Epigenetic Process; Excision; Flow Cytometry; Future; Gene Expression; Genetic Transcription; Goals; HIV; HIV Infections; Histone Deacetylation; In Vitro; Individual; Infection; Interleukin-15; Interruption; Intervention; Laboratories; Lead; Longevity; Lymphocyte Suppression; Macaca; Maintenance; Mediating; Methylation; Modeling; Molecular; Outcome; Phenotype; Play; Predisposition; Production; Role; SIV; Series; Shock; T-Lymphocyte Subsets; Testing; Therapeutic; Transcript; Viral; Viral reservoir; Viremia; Virus; Virus Replication; Work; antiretroviral therapy; base; combinatorial; cytokine; design; effective intervention; experimental study; high dimensionality; histone methylation; improved; in vitro Model; in vivo; innovation; novel; novel strategies; transcription factor; viral rebound; viral rescue

ABSTRACT: Current antiretroviral therapy (ART) does not eradicate HIV as shown by the rapid rebound of viremia upon treatment interruption. Rescuing viral gene expression in latently infected cells by compounds termed Latency Reversing Agents (LRA) is a strategy to reduce the virus reservoir in ART-treated HIV-infected individuals (i.e., shock & kill). HIV latency is triggered by mechanisms that lead to the silencing of virus expression including epigenetic DNA modification through histone methylation and deacetylation, limited availability of critical transcription factors and inefficient elongation of the nascent viral transcripts. Recent studies in ART-treated SIV- infected macaques demonstrated that depletion of CD8+ lymphocytes is followed by reactivation of virus production, and increased susceptibility to the LRA effect of the IL-15 superagonist ALT803. These results strongly suggest that CD8+ lymphocyte play an important yet understudied role in silencing HIV expression. In this proposal, we hypothesize that CD8+ T cells suppress HIV gene expression by promoting the establishment and maintenance of HIV latency, and that reversal of this effect may result in a significant amplification of the LRA effect of various compounds, thus impacting the reservoir size and stability in vivo. We will use innovative approaches to identify the CD8+ T cell mediated mechanisms of HIV silencing. In Aim 1, we will employ a novel model of HIV latency to characterize the role of CD8+ T cells in the establishment of HIV latency. We will quantify HIV gene expression in CD4+ T cell subsets and characterize the impact of CD8+ T cells on HIV latency establishment. In Aim 2, we will characterize the role of CD8+ T cells in the maintenance of HIV latency. Our in vitro model will allow us to define how CD8+ T cells maintain HIV latency during cytokine driven activation, proliferation or differentiation. In Aim 3, we will design a combinatorial anti-latency strategy that will eliminate most if not all latently infected cells. We will employ high dimensional flow cytometry to define alterations in phenotype associated with suppression and activation of latency reversal. Ultimately, we will characterize the role of CD8+ lymphocyte in counter-acting the latency reversal by specific LRAs (and LRA combinations) as a key step to develop effective interventions in which viral transcription in latently infected cells is rescued to reduce the reservoir size. The experiments described in this proposal will identify novel mechanisms of HIV silencing that can lead to the reactivation and subsequent elimination of most (if not all) latently infected cells. We propose that this overarching goal can be achieved through a step-wise scientific approach that includes in vitro and ex vivo studies that will address a newly-defined mechanism of HIV persistence. We believe that the work proposed in this application will allow us to understand the mechanisms by which CD8+ lymphocytes suppress virus transcription and ultimately promote HIV latency and persistence in ART-treated HIV-infected individuals, thus helping designing new approaches for HIV eradication.

抽象的： 当前的抗逆转录病毒疗法（ART）不会消除艾滋病毒，如病毒血症在 治疗中断。通过称为潜伏期的化合物在潜在感染细胞中挽救病毒基因表达 逆转剂（LRA）是减少经过艺术治疗的HIV感染个体（即， 震惊与杀戮）。 HIV潜伏期是由导致病毒表达沉默的机制引发的 通过组蛋白甲基化和脱乙酰化的表观遗传DNA修饰，关键的可用性有限 新生病毒转录本的转录因子和效率低下。最近关于艺术处理的SIV的研究 感染的猕猴表明CD8+淋巴细胞的耗竭后，病毒重新激活 生产，并提高对IL-15超级飞机ALT803 LRA效应的敏感性。这些结果 强烈建议CD8+淋巴细胞在沉默HIV表达中起重要但研究的作用。 在此提案中，我们假设CD8+ T细胞通过促进 艾滋病毒潜伏期的建立和维护以及这种影响的逆转可能会导致重大 各种化合物的LRA效应的扩增，从而影响体内储层大小和稳定性。我们 将使用创新方法来识别CD8+ T细胞介导的HIV沉默机制。在AIM 1中，我们 将采用一种新型的HIV潜伏期模型来表征CD8+ T细胞在艾滋病毒建立中的作用 潜伏期。我们将量化CD4+ T细胞子集中的HIV基因表达，并表征CD8+ T的影响 艾滋病毒潜伏期建立的细胞。在AIM 2中，我们将表征CD8+ T细胞在维护中的作用 艾滋病毒潜伏期。我们的体外模型将使我们能够定义CD8+ T细胞在细胞因子过程中如何保持HIV潜伏期 驱动的激活，增殖或分化。在AIM 3中，我们将设计一种组合反映策略 将消除大多数（如果不是全部）感染的细胞。我们将采用高维流式细胞仪来定义 与抑制和激活潜伏期逆转有关的表型的改变。最终，我们会的 表征CD8+淋巴细胞在通过特定LRA（和LRA 组合）作为开发有效干预措施的关键步骤，在该干预措施中，潜在感染细胞中的病毒转录 被救出以减少储层尺寸。该提案中描述的实验将确定新的机制 艾滋病毒沉默可能导致重新激活和随后消除大多数（如果不是全部）被延迟感染的 细胞。我们建议可以通过逐步的科学方法来实现这个总体目标 包括体外和离体研究，这些研究将解决新定义的HIV持久性机制。我们相信 本应用程序中提出的工作将使我们能够了解CD8+的机制 淋巴细胞抑制病毒转录，并最终促进ART治疗的HIV潜伏期和持久性 感染了艾滋病毒的人，从而帮助设计新的消除艾滋病毒方法。