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

超急性和急性艾滋病毒感染期间女性生殖道内的免疫反应和病毒动态

• 批准号: 10547981
• 负责人: Benjamin Joseph Read
• 金额: $ 6.72万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-01 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10547981
• 关键词: Acute; Address; Africa South of the Sahara; Antigen-Presenting Cells; Area; Biological Models; Biopsy; Blood; CD4 Positive T Lymphocytes; Cells; Clinical; Data; Dendritic Cells; Detection; Early treatment; Education; Environment; Epidemic; Ethics; Event; Experimental Models; Female; Female genitalia; Frequencies; Future; Genital; Genitalia; HIV; HIV Infections; Health; High Prevalence; Human; Image; Immune; Immune response; Immunofluorescence Immunologic; Immunologics; In Situ Hybridization; In Vitro; Incidence; Individual; Infection; Infection prevention; Inflammation; Inflammatory; Inflammatory Response; Intervention; Literature; Methods; Modeling; Morbidity - disease rate; Mucous Membrane; Participant; Pathogenesis; Phase; Plasma; Population; Prevalence; Process; Proteins; Province; Risk; Running; Sampling; Site; South Africa; Spatial Distribution; Structure; Systemic infection; T-Lymphocyte Subsets; Techniques; Testing; Therapeutic Intervention; Time; Tissues; Transcript; Ursidae Family; Validation; Viral; Viremia; Virus Diseases; Vulnerable Populations; acute infection; cervicovaginal; chemokine; chronic infection; cohort; cytokine; design; genital infection; human female; immunoregulation; improved; infection burden; insight; intervention effect; microscopic imaging; model design; mortality; nonhuman primate; pandemic disease; peripheral blood; prevent; programs; reproductive tract; response; single-cell RNA sequencing; systemic inflammatory response; therapeutic development; therapy development; transcriptome sequencing; transcriptomics; young woman

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.

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， 妇女然而，大多数关于女性生殖道（FGT）超急性和急性HIV感染的研究都是 在体外、离体或非人灵长类动物（NHP）模型中进行。因为 其中，在感染的最早阶段，人FGT中对HIV的免疫应答在很大程度上是不确定的 目前的模型系统在多大程度上再现了自然感染事件尚不清楚。重要的是， FGT自然感染HIV后第一批被感染的细胞仍未被确认，这是一个主要障碍 发展预防感染的干预措施。FRESH（女性崛起） 南非夸祖鲁-纳塔尔的教育、支持和健康）队列是一个长期运行的项目， 旨在识别自然的、超急性的HIV感染，并以纵向方式收集各种样本， 提供了一个前所未有的机会，以检查免疫和病毒动态的FGT期间， 超急性和急性HIV感染。通过利用已验证的尖端转录组技术 通过我们的小组，我们将能够阐明在感染的这一关键时期内FGT内的事件。 我们的总体假设是，FGT将有一个强烈的炎症反应，高峰早于 外周血中全身反应和早期炎症将主要由抗原呈递驱动 细胞（APC），特别是浆细胞样树突状细胞（pDC），而CD4+ T细胞亚群将承担最大的 生殖器感染的负担。在本提案的第一个目的中，我们将研究生殖器免疫应答， 使用纵向收集的大量细胞群体样本通过群体水平RNA测序检测HIV感染 从超急性和急性感染的FRESH参与者。我们还将进行单细胞转录组学 在检测到感染后的最早时间点分析生殖道细胞， 这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的免疫荧光（IF）成像进行补充 来自FRESH参与者的活检组织。在第二个目标中，我们将研究超急性的病毒动力学， 通过使用单细胞RNA测序和串联测序技术识别和可视化感染细胞， 原位杂交（ISH）和IF成像。此外，我们将研究目前的病毒准种的多样性， 在FGT和血浆中，以更好地确定局部感染灶的频率， FGT成功发展以建立全身感染。这些研究将首次使 明确概述在超急性和急性HIV感染期间发生的关键早期免疫和病毒事件 的人类FGT，允许验证现有的实验模型，并告知发展 预防艾滋病毒感染和传播的干预措施。