Evaluating macrophage antiviral immunity as a suppressive factor in SIV-M. tuberculosis co-infection

评估巨噬细胞抗病毒免疫作为 SIV-M 的抑制因素。

• 批准号: 10547182
• 负责人: Joshua T. Mattila
• 金额: $ 22.73万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-01 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10547182
• 关键词: Affect; Animals; Anti-Bacterial Agents; Anti-Retroviral Agents; Antibodies; Antimycobacterial Agents; Antiviral Response; Bacteria; CD4 Positive T Lymphocytes; Cell Communication; Cell Count; Cells; Clinical; Data; Development; Down-Regulation; Effector Cell; Experimental Models; Functional disorder; Future; Gene Expression; Gene Expression Profile; Genetic Transcription; Goals; Granuloma; HIV; HIV Infections; HIV vaccine; HIV/TB; Human; Image; Image Analysis; Immune system; Immunity; Immunobiology; Immunocompetent; In Vitro; Individual; Infection; Influenza; Interferon Type II; Interferons; Knowledge; Lead; Lesion; Link; Lung; Macaca; Machine Learning; Macrophage Activation; Mediating; Modeling; Monkeys; Mus; Mycobacterium tuberculosis; Normal Range; Opportunistic Infections; Organoids; Outcome; Pathogenesis; Pathology; Pathway interactions; Periodicity; Peripheral; Peripheral Blood Mononuclear Cell; Persons; Pharmaceutical Preparations; Population; Predisposition; Proteins; Research; Risk; SIV; Signal Transduction; Structure of parenchyma of lung; System; T-Lymphocyte; TNF gene; Testing; Tissues; Transcript; Tuberculosis; Viral load measurement; Viremia; Virus; Virus Diseases; Work; acute infection; antimicrobial; antiviral immunity; base; co-infection; cytokine; experience; high dimensionality; human model; immunosuppressive macrophages; improved outcome; in vivo; innovation; insight; macrophage; mouse model; mycobacterial; nonhuman primate; novel; pathogen; peripheral blood; preservation; programs; pulmonary granuloma; response; tuberculosis immunity

PROJECT SUMMARY: A third of the world’s population is infected with Mycobacterium tuberculosis (Mtb), the bacterium that causes tuberculosis (TB). The human immune system is highly successful at controlling Mtb and immunocompetent individuals who are infected have a 5-10% lifetime chance of experiencing active (symptomatic) TB. This level of protection is mediated by cytokine-based cell-cell communication in granulomas, the lesions associated with TB. Factors that dysregulate the protective Th1 responses, including downregulation of the macrophage activat- ing cytokines IFNγ and TNF, can promote active TB. HIV significantly increases the risk of developing TB and HIV-Mtb coinfected individuals experience a 5-15% annual risk of developing active TB and TB is a leading killer in this population. This elevated risk occurs in individuals who have normal range peripheral CD4+ T cell numbers and well controlled virus loads. Studies in SIV-Mtb coinfected nonhuman primates (NHPs), which are the best available model of HIV-Mtb coinfection, show that granulomas from coinfected and Mtb-only animals contain similar numbers of Mtb-specific T cells. These data suggest that virus-associated T cell-independent factors promote TB in SIV/HIV+ individuals. Macrophages may be the key to understanding the pathology of HIV-Mtb coinfection. Macrophages can support Mtb replication but are also the primary anti-Mtb effector cell in granulo- mas. Our preliminary data using RNAscope indicates that macrophages in NHP granulomas are infected with SIV and transcriptional analysis of granulomas from SIV+ and SIV- monkeys shows that coinfected granulomas have increased expression of type 1 interferon (IFN1) transcripts. IFN1s are induced by viral infection but are also associated with exacerbated TB, and based on these data, we hypothesize that viral infection induces IFN1- regulated immunity in granuloma macrophages and this suppresses macrophage antimycobacterial immunity, thus promoting TB. This hypothesis cannot be tested in humans and murine models are experimentally tractable but have significantly different immunobiology, HIV susceptibility, and TB presentation than humans. To over- come these obstacles, we will use highly multiplexed cyclic IHC and RNAscope on FFPE NHP granulomas to identify the relationship between IFN1, SIV infection, and macrophage anti-Mtb activity in SIV/Mtb coinfection. These studies will be done on a unique set of NHP granulomas with known Mtb and SIV loads, and we will use computational image analysis and machine learning to identify SIV+/- cells, how SIV affects cellular organization, IFN1 expression, and macrophage activation. To test our findings from these studies, we will use lung granuloma organoids composed of lung tissue from Mtb-infected macaques, in combination with fluorescent protein ex- pressing Mtb, SIV and IFN1 inhibition. These studies will identify relationships between SIV-regulated IFN1 ex- pression and Mtb survival in coinfected granulomas. Our long-term goal is to identify host-directed therapies to improve outcomes in HIV-Mtb coinfection and the mechanistic studies we are proposing with this innovative and translational system will produce new information on HIV-TB pathogenesis that contributes to this objective.

项目摘要： 世界三分之一的人口感染了结核分枝杆菌（MTB），这是引起的细菌 结核病（TB）。人类免疫系统在控制MTB和免疫能力方面非常成功 被感染的人有5-10％的终身机会体验活跃（症状）结核病。这个级别 保护的介导是由基于细胞因子的细胞细胞通信在颗粒中，与之相关的病变 TB。失调受保护的TH1反应的因素，包括巨噬细胞活动的下调 ING细胞因子IFNγ和TNF可以促进主动TB。艾滋病毒显着增加了发展结核病的风险 HIV-MTB共感染的个体的年每年出现活跃结核病和结核病的风险为5-15％ 在这个人群中。这种升高的风险发生在正常范围外围CD4+ T细胞数的个体中 和控制良好的病毒负荷。 SIV-MTB共同感染的非人类素数（NHP）的研究，这是最好的 HIV-MTB共感染的可用模型表明，共同感染和仅MTB动物的肉芽肿包含 相似数量的MTB特异性T细胞。这些数据表明与病毒相关的T细胞独立因素 在SIV/HIV+个体中促进结核病。巨噬细胞可能是理解HIV-MTB病理学的关键 共感染。巨噬细胞可以支持MTB复制，但也是颗粒中的主要抗MTB效应细胞 马斯。我们使用rnascope的初步数据表明，NHP肉芽瘤中的巨噬细胞感染 SIV+和SIV-猴子的颗粒的SIV和转录分析表明，共感染了肉芽肿 1型干扰素（IFN1）转录本的表达增加。 IFN1由病毒感染诱导，但 也与加重的结核病有关，并基于这些数据，我们假设病毒感染会引起IFN1- 肉芽肿巨噬细胞中受调节的免疫学，这抑制了巨噬细胞抗菌免疫学， 从而促进结核病。该假设不能在人类中进行检验，鼠模型是可以实验的 但是与人类具有明显不同的免疫生物学，HIV敏感性和结核病的表现。过度 - 出现这些障碍，我们将在FFPE NHP粒状瘤上使用高度多重的环状IHC和RNASCOPE 确定SIV/MTB共感染中IFN1，SIV感染和巨噬细胞抗MTB活性之间的关系。 这些研究将对具有已知MTB和SIV负载的独特NHP肉芽肿进行，我们将使用 计算图像分析和机器学习以识别SIV +/-细胞，SIV如何影响细胞组织， IFN1表达和巨噬细胞激活。为了从这些研究中测试我们的发现，我们将使用肺肉芽肿 由MTB感染猕猴组成的肺组织组成的类器官，结合荧光蛋白Ex- 按下MTB，SIV和IFN1抑制。这些研究将确定SIV调节的IFN1 ex- 固定颗粒。我们的长期目标是确定宿主指导的疗法 改善HIV-MTB共感染的结果以及我们通过这种创新性和 翻译系统将产生有关HIV-TB发病机理的新信息，这有助于这一目标。