The effect of HIV infection on CD4 T cell immunity to Mycobacterium tuberculosis

项目概要/摘要 结核分枝杆菌（Mtb）感染导致1000万例活动性结核（TB）疾病 每年都发生，并且是由于单一感染剂导致死亡的主要原因。虽然保护性的相关因素 对结核分枝杆菌免疫力没有明确定义，与人类免疫缺陷病毒（HIV）的合并感染是一个重要的 发展活动性结核病的危险因素。此外，结核病风险的增加在以下人群中是显而易见的： HIV血清转换的第一年，表明对Mtb的免疫损害发生在HIV感染后早期。 尽管有令人信服的证据表明HIV感染与CD 4 T细胞功能障碍有关，但HIV感染的分子机制可能与CD 4 T细胞功能障碍有关。 HIV感染损害CD 4 T细胞对Mtb应答的机制尚不清楚。本提案的重点 确定HIV感染和治疗对表型、功能、表观基因组和 CD 4 T细胞对Mtb应答的转录组。我们将利用从一个独特的，嗯- 在肯尼亚的蒙巴萨，对艾滋病毒感染高危妇女进行纵向队列研究， 假设（i）HIV感染改变了CD 4 T细胞的表观基因组和转录组， 分化功能障碍的CD 4 T细胞;和（ii）早期开始抗逆转录病毒治疗（ART），在HIV-1感染之前， 诱导的CD 4 T细胞缺陷，保留了CD 4 T细胞应答的分子程序和功能能力 这可能有助于增强对结核分枝杆菌的免疫控制。我们将使用纵向样本进行高- 三维流式细胞术，以确定总的和Mtb特异性CD 4 T细胞的表型和功能， 在每个时间点，我们还将定义染色质， 总CD 4 T细胞群体的可及性景观、DNA甲基化组和转录组。最后，我们将 使用尖端的单细胞RNA测序技术来确定HIV感染和治疗对 Mtb特异性CD 4 T细胞的转录状态。这些研究将产生前所未有的新见解， HIV感染中CD 4 T细胞的表型、功能、转录组学和表观遗传学特征。而且我们 全基因组研究将定义HIV感染中CD 4 T细胞的分子程序，并可能识别 新的基因、途径和基于转录因子的调控模块，可以利用它们来增强 持久的CD 4 T细胞介导的Mtb控制。

Project Summary / Abstract Infection with Mycobacterium tuberculosis (Mtb) results in 10 million cases of active tuberculosis (TB) disease each year and is the leading cause of death due to a single infectious agent. Although the correlates of protective immunity to Mtb are not clearly defined, co-infection with human immunodeficiency virus (HIV) is a significant risk factor for development of active TB disease. Moreover, the increased risk of TB disease is evident within the first year of HIV seroconversion, suggesting that impairment of immunity to Mtb occurs early after HIV infection. Although there is compelling evidence that HIV infection is associated with CD4 T cell dysfunction, the molecular mechanisms by which HIV infection impairs CD4 T cell responses to Mtb are unknown. The focus of this proposal is to determine the effect of HIV infection and treatment on the phenotype, function, epigenome and transcriptome of CD4 T cell responses to Mtb. We will leverage blood samples collected from a unique, well- characterized longitudinal cohort of women at high-risk for HIV infection in Mombasa, Kenya to test the hypotheses that (i) HIV infection modifies the epigenome and transcriptome of CD4 T cells to promote differentiation of dysfunctional CD4 T cells; and (ii) early initiation of antiretroviral therapy (ART), prior to HIV- induced CD4 T cell deficiency, preserves the molecular program and functional capacity of CD4 T cell responses that may contribute to enhanced immune control of Mtb. We will use longitudinal samples to conduct high- dimensional flow cytometry to define the phenotype and function of total and Mtb-specific CD4 T cells before and after HIV infection, and before and after ART. At each time point, we will also define the chromatin accessibility landscape, DNA methylome, and transcriptome of the total CD4 T cell population. Lastly, we will use cutting edge, single-cell RNA-sequencing to determine the effect of HIV infection and treatment on the transcriptional state of Mtb-specific CD4 T cells. These studies will generate unprecedented, novel insights into the phenotypic, functional, transcriptomic, and epigenetic profiles of CD4 T cells in HIV infection. Moreover, our genome-wide studies will define the molecular program of CD4 T cells in HIV infection and will potentially identify novel genes, pathways, and transcription factor-based regulatory modules that can be leveraged to enhance durable CD4 T cell-mediated control of Mtb.