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

摘要摘要 有大量证据表明，在病毒学抑制的艾滋病毒携带者(VsPWH)中，艾滋病毒仍然存在 血液和组织中的单核细胞和巨噬细胞。然而，尽管单核细胞和 有限的研究表明，巨噬细胞对艾滋病毒储备库的大小有贡献，并可能破坏基于治疗的努力 正在研究单核细胞和巨噬细胞内的艾滋病毒是否可以被重新激活以产生 VsPWH中的病毒。此外，尽管已知的基于性别的CD4储存库和免疫学差异 对艾滋病毒的反应，目前还没有报道评估髓系细胞的性别差异 (单核/巨噬细胞)储存于vsPWH。这项提议的长期目标是确定髓系细胞 蓄水池是以治愈为主的努力的重大问题。总体目标是：(1)确定是否有 髓系储存库的性别差异，(2)决定了血液中髓系储存库的稳定性 VsPWH和SIV-ART猕猴的单核细胞来源的巨噬细胞(MDM)和脑(CNS)以及(3)进一步 开发一种易于部署的方法来评估vsPWH中的髓系储存库。这一点的中心假设是 工作是，在男性和女性MDM和CNS可重新激活的储集层中将有显著差异 在HIV和SIV中，这些髓系储存库在ART抑制过程中将是稳定的。这个 拟议研究的基本原理是描绘MDM和CNS髓系的大小和稳定性 水库，以及潜在的基于性别的差异，将允许开发基于治愈的策略 适当地针对携带艾滋病毒的男性和女性的髓系储存库。核心假设将通过以下方式进行检验 追求两个具体目标：1.确定vsPWH中MDM HIV感染者的性别差异，以及2. 确定SIV-ART猕猴中MDM和CNS储存区的性别差异。这些目标将 利用新的HIV或SIV特异性技术，单核细胞来源的巨噬细胞定量病毒副产物 MDM-QVOA(MDM-QVOA)、CNS巨噬细胞定量病毒生长试验(CNS-QVOA)和髓系 改进的完整前病毒DNA测定法(MIPDA)，用于测量具有复制能力和完整的病毒库 MDM(vsPWH和SIV-ART模型)和CNS(SIV-ART猕猴)。本研究方案具有创新性。 因为它关注的是复制能力而不是DNA，它将阐明潜在的基于性别的差异 并解决了vsPWH和SIV-ART猕猴髓系储存库的稳定性问题。最后，这项建议是 意义重大，因为它将提供对单核细胞和巨噬细胞储存库的基本见解，并有助于 确定HIV是否在髓系起源的细胞中建立了真正的潜伏期。归根结底，这样的知识有可能 改变艾滋病毒携带者的基于治愈的努力和治疗方法，就像目前的大多数努力一样 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.