HIV-a Persistence in Myeloid Cell Reservoirs

项目总结： 随着艾滋病毒/艾滋病研究领域开始努力治愈艾滋病毒-1感染，洞察到 治愈感染是必不可少的。虽然这方面的大部分注意力都集中在CD4+T细胞储存库上， 人们对组织巨噬细胞是否为HIV-1感染者体内的蓄水池的关注要少得多 抑制性抗逆转录病毒疗法(ART)事实上，巨噬细胞储存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完全是感染的吞噬细胞的结果 CD4+T细胞(Calantone等人，2014年)。因此，关于髓系细胞储存库的研究正在变得默默无闻。 巨噬细胞的感染只能由能够使用低水平CD4的HIV-1变异株启动 在细胞表面。因此，我们建议通过巨噬细胞嗜性的存在来揭示髓系细胞储存库。 来自ART抑制个体淋巴组织的DNA和细胞相关RNA的包膜 从从接受分析治疗中断的个体获得的最早复发的病毒 (ATI)。这种不偏不倚的方法不依赖于将髓系细胞纯化到同质性，也不是混淆的 通过吞噬的CD4+T细胞的存在。为了进一步证实髓系细胞是功能储存库，我们将 试图直接证明巨噬细胞嗜好、复制能力强的病毒在可访问的、 巨噬细胞丰富的解剖小室。 具体来说，我们建议： 目的1：从ART抑制的淋巴组织(LN、GALT)中鉴定巨噬细胞嗜性变异体 个体和ATI后早期复发的变种。我们将采用半高吞吐量取向 一种能够识别低频率、嗜巨噬细胞的病毒以推断巨噬细胞的存在的方法 水库。 目的2：确定肺泡巨噬细胞是否支持HIV-1在抑制状态下持续存在 艺术。肺泡巨噬细胞病毒群与CD_4~+T-T细胞的亲和性及系统发育关系 将对来自无菌个体的细胞进行评估，并从肺泡中分离出嗜巨噬细胞病毒 巨噬细胞体外培养 这些研究应该第一次为组织巨噬细胞中存在病毒储存库提供确凿证据。 因此，为设计清除巨噬细胞储存库的策略提供催化剂并增加 治愈感染的可能性。

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.