Defining sex-based differences in the replication-competent myeloid reservoir in virally suppressed people with HIV and SIV-infected ART-suppressed macaques

摘要 在病毒学抑制的HIV感染者（vsPWH）中有大量证据表明， 单核细胞和巨噬细胞。然而，尽管单核细胞和 巨噬细胞有助于艾滋病毒水库的大小，并可能破坏基于治疗的努力，有限的研究 目前正在研究单核细胞和巨噬细胞内的HIV是否可以被重新激活， vsPWH中的病毒。此外，尽管已知在CD 4储库和免疫学方面存在性别差异， 对HIV的反应，目前还没有研究报告评估骨髓中基于性别的差异， （单核细胞/巨噬细胞）储库。这项提案的长期目标是确定髓系 储层是基于治愈的努力的一个重要关注点。总体目标是：（1）确定是否存在 基于性别的骨髓储库差异，（2）决定骨髓储库在血液中的稳定性 （3）进一步研究vsPWH和SIV-ART猕猴的单核细胞衍生的巨噬细胞（MDM）和脑（CNS）的细胞毒性， 开发一种易于部署的方法来评估vsPWH中的骨髓储库。这个问题的核心假设是 这项研究的结果是，男性和女性的MDM和CNS可再活化储库存在显著差异， 在HIV和SIV中，这些骨髓储库在整个ART抑制过程中将是稳定的。的 拟议研究的基本原理是描述MDM和CNS髓样细胞的大小和稳定性， 水库，和潜在的性别差异，将允许开发基于治疗的战略， 适当地针对艾滋病毒感染者的骨髓储库。中心假设将通过以下方式进行检验： 具体目标有两个：1.定义vsPWH中MDM HIV储库中基于性别的差异，以及2. 定义SIV-ART猕猴中MDM和CNS储库的性别差异。这些目标将 利用新的HIV或SIV特异性技术， 通过MDM-QVOA、CNS巨噬细胞定量病毒生长测定（CNS-QVOA）和髓样细胞定量病毒生长测定（MDM-QVOA）， 适应的完整前病毒DNA测定法（mIPDA），以测量 MDM（vsPWH和SIV-ART模型）和CNS（SIV-ART猕猴）。这一研究建议具有创新性 因为它关注的是复制能力而不是DNA，将阐明潜在的性别差异， 并解决vsPWH和SIV-ART猕猴中骨髓储库的稳定性。最后，这一建议是 意义重大，因为它将提供对单核细胞和巨噬细胞储库的基本见解，并有助于 确定HIV是否在骨髓来源的细胞中建立真正的潜伏期。最终，这些知识有可能 改变男性和女性艾滋病毒感染者基于治愈的努力和治疗方法，因为目前的大多数努力都是 完全集中在T细胞上

SUMMARY ABSTRACT There is substantial evidence in virologically-suppressed people with HIV (vsPWH) that HIV persists in monocytes and macrophages from blood and tissues. However, despite the likelihood that monocytes and macrophages contribute to the size of the HIV reservoir, and may derail cure-based efforts, limited studies exist investigating whether HIV within monocytes and macrophages can be reactivated to produce functional virus in vsPWH. Additionally, despite known sex-based differences in the CD4 reservoir and immunological response to HIV, there are no reported studies that have assessed sex-based differences in the myeloid (monocyte/macrophage) reservoir in vsPWH. The long-term goal of this proposal is to determine if the myeloid reservoir is a significant concern for cure-based efforts. The overall objectives are to (1) determine if there are sex-based differences in the myeloid reservoir, (2) determine the stability of the myeloid reservoir in the blood (monocyte derived macrophage, MDM) and brain (CNS) of vsPWH and SIV-ART macaques and (3) further develop an easily deployable method to assess the myeloid reservoir in vsPWH. The central hypothesis of this work is that there will be significant differences in the male and female MDM and CNS reactivatable reservoirs in both HIV and SIV, and that these myeloid reservoirs will be stable throughout ART suppression. The rationale for the proposed study is that delineating the size and stability of the MDM and CNS myeloid reservoirs, and potential sex-based differences, will allow for the development of cure-based strategies that appropriately target the myeloid reservoir in men and women with HIV. The central hypothesis will be tested by pursing two specific aims: 1. Define sex-based differences in the MDM HIV reservoir in vsPWH, and 2. Define sex-based differences in the MDM and CNS reservoirs in SIV-ART macaques. These aims will utilize novel HIV or SIV specific techniques, the monocyte derived macrophage quantitative viral outgrowth assay (MDM-QVOA), the CNS macrophage quantitative viral outgrowth assay (CNS-QVOA) and a myeloid adapted intact proviral DNA assay (mIPDA), to measure the replication-competent and intact viral reservoirs in MDM (vsPWH and SIV-ART model) and the CNS (SIV-ART macaques). This research proposal is innovative because it focuses on replication-competence rather than DNA, will elucidate potential sex-based differences and address the stability of myeloid reservoirs in vsPWH and SIV-ART macaques. Finally, this proposal is significant because it will provide essential insights into the monocyte and macrophage reservoir and help to determine if HIV establishes true latency in cells of myeloid origin. Ultimately, such knowledge has the potential to alter cure-based efforts and therapeutics in men and women with HIV as the majority of current efforts are entirely T cell focused.