The impact of TB co-infection on the reservoir of persistent HIV in vivo

 描述（由申请人提供）：最近的研究强调了靶向持久性HIV储库以实现功能性治愈的重要性。髓系细胞谱系在维持持久性病毒储库中的作用知之甚少。结核分枝杆菌（Mtb）直接感染巨噬细胞，并与增加的HIV病毒载量和更差的临床结果，尽管治疗两种病原体。这与其他机会性感染不同，如肺孢子虫肺炎（PCP），其中循环病毒载量的增加对治疗迅速作出反应。因此，Mtb感染可能促进HIV骨髓储库的持续存在甚至扩大。我们的目的是描述在体内活动性结核病（TB）背景下HIV的骨髓储库。为此，我们计划在TB肉芽肿相关巨噬细胞和肺泡巨噬细胞内目视检测HIV DNA和RNA，以评估HIV和Mtb的DNA或RNA的细胞共定位，并辨别HIV和Mtb治疗后是否存在HIV的这种假定骨髓储库的消退。我们假设，活动性或近期活动性TB将与持续的HIV感染储库相关，尽管对HIV和Mtb进行了治疗; HIV和Mtb将在巨噬细胞内共定位; TB肉芽肿相关巨噬细胞将更频繁地含有HIV DNA和RNA。证明组织巨噬细胞内存在可量化的结核分枝杆菌依赖性HIV储库，尽管进行了HIV和结核分枝杆菌治疗，但这种储库仍然存在，这对于未来旨在根除HIV的战略非常重要，特别是在世界上结核病猖獗的许多地方。我们组建了一个由结核病研究人员和艾滋病毒免疫学家组成的多学科团队，他们在艾滋病毒检测和临床研究设计的超灵敏方法方面具有专业知识。我们的工作利用了由NIH和Wellcome Trust资助的现有研究队列，从患有和不患有活动性TB的HIV感染受试者中获取生物库福尔马林固定石蜡包埋组织，并前瞻性地从患有HIV和活动性TB或HIV和PCP的受试者中收集支气管肺泡灌洗液（BAL）和外周血。样本将用于：（1）检测和定量HIV的组织骨髓储库;（2）评估HIV与细胞内Mtb的细胞共定位;（3）比较开始ART和最近治疗活动性TB或急性PCP的HIV感染受试者中肺泡巨噬细胞HIV储库的持久性。如果这项探索性研究揭示了结核分枝杆菌特异性骨髓HIV储库的存在，我们将进行更全面的前瞻性研究，以调查结核分枝杆菌感染可能维持持续HIV储库的机制，即使在抗逆转录病毒治疗的情况下。

 DESCRIPTION (provided by applicant): Recent studies highlight the importance of targeting the persistent HIV reservoir to attain a functional cure. The role of the myeloid cell lineage in maintaining a persistent viral reservoir is poorly understood. Mycobacterium tuberculosis (Mtb) directly infects macrophages, and is associated with increased HIV viral loads and worse clinical outcomes despite treatment for both pathogens. This is unlike other opportunistic infections, such as Pneumocystis Pneumonia (PCP), in which increases in circulating viral loads quickly respond to treatment. Thus, Mtb infection may facilitate the persistence and even expansion of a myeloid reservoir of HIV. Our objective is to describe the myeloid reservoir of HIV in the setting of active tuberculosis (TB) in vivo. To that end, we plan to visually detect HIV DNA and RNA within TB granuloma-associated macrophages and alveolar macrophages, to assess cellular co-localization of DNA or RNA from HIV and Mtb, and to discern whether there is resolution of this putative myeloid reservoir of HIV after treatment for HIV and Mtb. We hypothesize that active or recently active TB will be associated with a sustained reservoir of HIV infection despite treatment for HIV and Mtb; that HIV and Mtb will co-localize within macrophages; and that TB granuloma-associated macrophages will more frequently contain HIV DNA and RNA. Demonstration of a quantifiable Mtb-dependent HIV reservoir within tissue macrophages that persists despite HIV and Mtb treatment will be important for future strategies aimed at HIV eradication, particularly in those many places around the world where TB is rampant. We have assembled a multidisciplinary team of TB researchers and HIV immunologists with expertise in ultra-sensitive methods of HIV detection and clinical study design. Our work leverages existing research cohorts funded by the NIH and the Wellcome Trust, accessing biobanked formalin-fixed paraffin- embedded tissue from HIV-infected subjects with and without active TB, and prospectively collecting bronchoalveolar lavage (BAL) and peripheral blood from subjects with HIV and active TB or HIV and PCP. Samples will be used to: (1) detect and quantify the tissue myeloid reservoir of HIV; (2) assess cellular co- localization o HIV with intracellular Mtb; (3) compare persistence of the alveolar macrophage HIV reservoir in HIV-infected subjects started on ART and recently treated for active TB or acute PCP. Should this exploratory study reveal the existence of an Mtb-specific myeloid reservoir of HIV, we will pursue more comprehensive prospective studies to investigate the mechanisms by which Mtb infection might sustain the persistent HIV reservoir, even in the face of antiretroviral therapy.