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。

基本信息

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
项目类别：
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

项目摘要

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.

项目概要：世界上三分之一的人口感染了结核分枝杆菌 (Mtb)，这种细菌会导致结核病 (TB) 人类免疫系统在控制 Mtb 和免疫功能方面非常成功。感染者终生有 5-10% 的机会罹患活动性（有症状）结核病。保护作用是由肉芽肿中基于细胞因子的细胞通讯介导的，肉芽肿是与 TB。保护性 Th1 反应失调的因素，包括巨噬细胞激活的下调。细胞因子 IFNγ 和 TNF 可以促进活动性结核病，显着增加患结核病的风险。 HIV-Mtb 双重感染者每年患活动性结核病的风险为 5-15%，结核病是主要杀手在这一人群中，外周 CD4+ T 细胞数量处于正常范围的个体会出现这种较高的风险。以及对 SIV-Mtb 共感染的非人灵长类动物 (NHP) 的病毒载量控制良好的研究，这是最好的。 HIV-Mtb 合并感染的现有模型表明，来自合并感染和仅 Mtb 动物的肉芽肿含有 Mtb 特异性 T 细胞数量相似，这些数据表明病毒相关 T 细胞独立因素。巨噬细胞可能是了解 HIV-Mtb 病理学的关键。巨噬细胞可以支持 Mtb 复制，但也是颗粒细胞中主要的抗 Mtb 效应细胞。我们使用 RNAscope 的初步数据表明 NHP 肉芽肿中的巨噬细胞被感染对 SIV+ 和 SIV- 猴的肉芽肿进行 SIV 和转录分析表明，共感染的肉芽肿 1 型干扰素 (IFN1) 转录物的表达增加，但 IFN1 是由病毒感染诱导的。也与结核病恶化有关，根据这些数据，我们发现病毒感染会诱导 IFN1- 调节肉芽肿巨噬细胞的免疫，从而抑制巨噬细胞的抗分枝杆菌免疫，从而促进结核病的发生，这一假设无法在人类身上进行检验，而小鼠模型在实验上是可以处理的。但与人类相比，其免疫生物学、艾滋病毒易感性和结核病表现存在显着差异。面对这些障碍，我们将在 FFPE NHP 肉芽肿上使用高度多重循环 IHC 和 RNAscope，以确定 SIV/Mtb 共感染中 IFN1、SIV 感染和巨噬细胞抗 Mtb 活性之间的关系。这些研究将在一组具有已知 Mtb 和 SIV 负荷的独特 NHP omas 上进行，我们将使用计算图像分析和机器学习来识别 SIV+/- 细胞，SIV 如何影响细胞组织，为了测试我们的研究结果，我们将使用肺肉芽肿。由感染 Mtb 的猕猴的肺组织组成的类器官，结合荧光蛋白 ex- 这些研究将确定 SIV 调节的 IFN1 ex-之间的关系。我们的长期目标是确定针对宿主的治疗方法。改善 HIV-Mtb 合并感染的结果以及我们提出的这一创新和机制研究转化系统将产生有关艾滋病毒结核病发病机制的新信息，有助于实现这一目标。