Immune response and viral dynamics within the female genital tract during hyperacute and acute HIV infection

项目摘要 艾滋病毒仍然是一种全球流行病，在流行率最高的地区，年轻人感染比例不成比例。 女人。然而，大多数关于女性生殖道的超急性和急性艾滋病毒感染(FGT)的研究是 由于实际和伦理方面的考虑，在体外、体外或非人类灵长类动物(NHP)模型中进行。因为 其中，人类FGT在感染的最早阶段对HIV的免疫反应很大程度上是未知的 目前的模型系统在多大程度上重述了自然感染事件尚不清楚。关键是， FGT自然感染HIV后感染的第一批细胞仍未确定，这构成了一个主要障碍 朝着预防感染的干预措施的发展。新鲜的(女性崛起通过 教育、支持和卫生)在南非夸祖鲁-纳塔尔的队列是一个长期运行的计划，是 旨在识别自然的、超急性的艾滋病毒感染，并纵向收集各种样本， 为研究FGT的免疫和病毒动力学提供了一个前所未有的机会 超急性和急性艾滋病毒感染。通过利用已被验证的尖端转录技术 通过我们的小组，我们将能够阐明在这一关键的感染时间段内FGT内发生的事件。 我们的总体假设是，FGT将有强烈的炎症反应，其峰值早于 外周血液中的全身反应和早期炎症将主要由抗原提呈驱动 细胞(APC)，特别是浆细胞样树突状细胞(PDCs)，而CD4+T细胞亚群将承担最大 生殖器感染的负担。在这项提案的第一个目标中，我们将研究生殖器的免疫反应。 使用纵向收集的大量细胞种群样本进行种群水平的RNA测序来实现HIV感染 来自极度尖锐和急性感染的新鲜参与者。我们还将进行单细胞转录 从生殖道细胞分析最早的时间点检测感染情况进行调查 哪些特定的细胞亚群负责炎症和免疫调节。这些分析 将通过Luminex细胞因子研究和宫颈阴道免疫荧光(IF)成像进行补充 新鲜受试者的活检组织。在第二个目标中，我们将研究超急性肺炎的病毒动力学。 用单细胞RNA测序和串联技术识别和可视化感染细胞的生殖器HIV感染 原位杂交(ISH)和IF成像。此外，我们还将研究目前存在的病毒准物种的多样性。 在FGT和在血浆中的这个时间点，以更好地确定局部感染灶在 FGT成功地发展为建立系统性感染。拟议的研究将首次使 明确概述在超急性和急性艾滋病毒感染期间发生的关键早期免疫和病毒事件 人类FGT，允许验证现有的实验模型，并通知开发 预防艾滋病毒感染和传播的干预措施。

Project Summary HIV remains a global epidemic and, in the areas of greatest prevalence, disproportionately infects young women. However, most studies of hyperacute and acute HIV infection of the female genital tract (FGT) are conducted in in vitro, ex vivo, or nonhuman primate (NHP) models due to practical and ethical concerns. Because of this, the immune response to HIV in the human FGT during the earliest stages of infection is largely undefined and the degree to which current model systems recapitulate the events of natural infection is unclear. Critically, the first cells to be infected following natural HIV infection of the FGT remain unidentified, creating a major hurdle toward the development of interventions effect at preventing infection. The FRESH (Females Rising through Education, Support, and Health) cohort in KwaZulu-Natal, South Africa, is a long-running program that is designed to identify natural, hyperacute HIV infection and collect a variety of samples in a longitudinal manner, presenting an unprecedented opportunity to examine the immune and viral dynamics of the FGT during hyperacute and acute HIV infection. By utilizing cutting-edge transcriptomic techniques that have been validated by our group, we will be able to elucidate the events within the FGT during this critical time period of infection. Our overall hypothesis is that the FGT will have a strong inflammatory response that peaks earlier than the systemic response in peripheral blood and that early inflammation will primarily be driven by antigen-presenting cells (APCs), particularly plasmacytoid dendritic cells (pDCs), while CD4+ T cell subsets will bear the greatest burden of genital infection. In the first Aim of this proposal, we will investigate the immune response to genital HIV infection via population-level RNA sequencing using longitudinally collected bulk cellular population samples from hyperacutely and acutely infected FRESH participants. We will also conduct a single cell transcriptomic analysis of cells from the genital tract at the earliest time point following the detection of infection to investigate which specific cellular subpopulations are responsible for inflammation and immune modulation. These analyses will be supplemented by Luminex cytokine studies and immunofluorescence (IF) imaging of cervicovaginal biopsy tissues from FRESH participants. In the second Aim, we will investigate the viral dynamics of hyperacute genital HIV infection by identifying and visualizing infected cells using single cell RNA sequencing and tandem in situ hybridization (ISH) and IF imaging. Additionally, we will examine the diversity of viral quasispecies present in the FGT and in plasma at this time point to better ascertain the frequency with which local foci of infection in the FGT successfully develop to establish systemic infection. The proposed studies will for the first time enable a clear outline of key early immunological and viral events that occur during hyperacute and acute HIV infection of the human FGT, allowing for the validation of existing experimental models and informing the development of interventions to prevent HIV infection and spread.