HIV-a Persistence in Myeloid Cell Reservoirs

HIV-a 在骨髓细胞库中的持续存在

• 批准号: 9204094
• 负责人: Mario Stevenson
• 金额: $ 19.19万
• 依托单位: UNIVERSITY OF MIAMI SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-07-01 至 2018-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9204094
• 关键词: AIDS/HIV problem; Alveolar; Alveolar Macrophages; Anatomy; Antiviral Agents; Attention; Biological Assay; CD4 Positive T Lymphocytes; Cell surface; Cells; DNA; Evaluation; Frequencies; Genome; Gut associated lymphoid tissue; HIV; HIV-1; Individual; Infection; Interruption; Lung; Lymphoid; Lymphoid Tissue; Macaca; Modeling; Myeloid Cells; Phagocytosis; Phylogenetic Analysis; Plasma; Play; Population; Property; Proviruses; RNA; Research; Role; SIV; Sampling; Surveys; Time; Tissues; Tropism; Variant; Viral; Viral Genome; Viral reservoir; Viremia; Virus; Virus Diseases; Work; antiretroviral therapy; catalyst; cell envelope; deep sequencing; design; digital; falls; insight; lymph nodes; macrophage; memory CD4 T lymphocyte; viral DNA; viral RNA; virus tropism

Project Summary: As the HIV/AIDS research field embarks on an endeavor to cure HIV-1 infection, insight into the obstacles to curing infection are essential. While most of the attention in this regard has focused on CD4+ T-cell reservoirs, far less attention has focused on whether tissue macrophages are a reservoir in HIV-1-infected individuals on suppressive antiretroviral therapy (ART). Indeed, the question of macrophage reservoirs has been polarized by recent work arguing that viral DNA in macrophages is exclusively a consequence of phagocytosis of infected CD4+ T-cells (Calantone et al., 2014). As a result, research on myeloid cell reservoirs is falling into obscurity. Infection of macrophages can only be initiated by HIV-1 variants that have the ability to use low levels of CD4 on the cell surface. We therefore propose to reveal myeloid cell reservoirs by the presence of macrophage-tropic envelopes from DNA and cell-associated RNA obtained from lymphoid tissues of ART-suppressed individuals and from the earliest recrudescing viruses obtained from individuals undergoing analytic treatment interruption (ATI). This unbiased approach does not rely on purification of myeloid cells to homogeneity nor is it obfuscated by the presence of phagocytosed CD4+ T-cells. To further validate myeloid cells as functional reservoirs, we will attempt to directly demonstrate the existence of macrophage-tropic, replication-competent virus in an accessible, macrophage-rich anatomic compartment. Specifically, we propose to: Aim 1: Identify macrophage-tropic variants in lymphoid tissue (LN, GALT) from ART-suppressed individuals and in early recrudescing variants following ATI. We will employ a semi-high throughput tropism assay, capable of identifying low-frequency, macrophage-tropic viruses to infer the existence of a macrophage reservoir. Aim 2: Establish whether alveolar macrophages support HIV-1 persistence in individuals on suppressive ART. Tropism and phylogenetic relationships between virus populations in alveolar macrophages and CD4+ T- cells from aviremic individuals will be assessed as well as isolation of macrophage-tropic virus from alveolar macrophages ex vivo These studies should, for the first time, provide definitive evidence for a viral reservoir in tissue macrophages and as such, provide the catalyst for the design of strategies to clear macrophage reservoirs and increase the likelihood of curing infection.

项目摘要： 随着艾滋病毒/艾滋病研究领域开始治愈HIV-1感染的努力，深入了解障碍 固化感染至关重要。尽管这方面的大多数关注都集中在CD4+ T细胞储层上，但 注意力较少的集中于组织巨噬细胞是否是HIV-1感染者的储层 抑制性抗逆转录病毒疗法（ART）。实际上，巨噬细胞储藏的问题已被极化 最近的工作认为，巨噬细胞中的病毒DNA仅是感染的吞噬作用的结果 CD4+ T细胞（Calantone等，2014）。结果，对髓样细胞储存库的研究陷入了晦涩之处。 巨噬细胞的感染只能由具有使用低水平CD4的HIV-1变体引发 在细胞表面。因此，我们建议通过存在巨噬细胞 - 循环透露髓样细胞储存剂 从Art抑制个体的淋巴组织获得的DNA和与细胞相关的RNA的信封 以及从接受分析治疗中断的个体获得的最早的重现病毒 （ati）。这种无偏的方法不依赖于髓样细胞的纯化，也不依赖于混淆。 通过吞噬的CD4+ T细胞的存在。为了进一步验证髓样细胞为功能储层，我们将 试图直接证明在可访问的，可访问的巨噬细胞效率，复制能力的病毒中存在 富含巨噬细胞的解剖室。 具体来说，我们建议： AIM 1：从Art抑制的 在ATI之后，个人和早期概述了变体。我们将采用半高的吞吐量 测定，能够鉴定低频，巨噬细胞 - 冰球病毒来推断巨噬细胞的存在 水库。 目标2：确定肺泡巨噬细胞是否支持抑制性的个体的HIV-1持久性 艺术。肺泡巨噬细胞中病毒种群与CD4+ T-的病毒群体之间的热爱和系统发育关系 将评估来自恶毒个体的细胞，并从肺泡中分离巨噬细胞 - 热带病毒 巨噬细胞离体 这些研究应首次为组织巨噬细胞中病毒储存库提供明确的证据 因此，为设计清除巨噬细胞库的策略设计并增加了催化剂 治愈感染的可能性。