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.

项目概要： 随着艾滋病毒/艾滋病研究领域开始奋进治愈HIV-1感染，深入了解艾滋病毒/艾滋病治疗的障碍， 治疗感染是必不可少的。虽然这方面的大多数注意力集中在CD 4 + T细胞库上， 人们对组织巨噬细胞是否是HIV-1感染者的储存库的关注要少得多。 抑制性抗逆转录病毒治疗（ART）。事实上，巨噬细胞储库的问题已经两极分化， 最近的研究认为，巨噬细胞中的病毒DNA完全是感染细胞吞噬的结果， CD 4 + T细胞（Calantone等人，2014年）。因此，对骨髓细胞储库的研究正陷入模糊。 巨噬细胞的感染只能由具有使用低水平CD 4的能力的HIV-1变体启动 在细胞表面。因此，我们建议通过巨噬细胞趋化因子的存在来揭示骨髓细胞储库。 来自ART抑制个体淋巴组织的DNA和细胞相关RNA的包膜 以及从经历分析治疗中断的个体获得的最早复发病毒 （ATI）。这种无偏倚的方法不依赖于骨髓细胞的纯化，也不混淆 通过吞噬的CD 4 + T细胞的存在。为了进一步验证骨髓细胞作为功能性储库，我们将 试图直接证明嗜巨噬细胞的存在，复制能力的病毒在一个可访问的， 巨噬细胞丰富的解剖室。 具体而言，我们建议： 目的1：鉴定ART抑制的淋巴组织（LN，GALT）中的巨噬细胞变体 个体和ATI后的早期复发变体。我们将采用半高通量取向 一种能够鉴定低频率嗜巨噬细胞病毒以推断巨噬细胞存在的检测方法 水库 目的2：确定肺泡巨噬细胞是否支持HIV-1在个体中的持续存在， 条病毒在肺泡巨噬细胞和CD 4 + T细胞中的嗜性和系统发育关系 将评估来自病毒血症个体的细胞以及从肺泡巨噬细胞嗜性病毒的分离。 离体巨噬细胞 这些研究应该是第一次为组织巨噬细胞中的病毒储库提供明确的证据 因此，为设计清除巨噬细胞储库和增加巨噬细胞增殖的策略提供了催化剂。 治愈感染的可能性。