A mechanistic understanding of tuberculosis progression through bacterial reporter strains

通过细菌报告菌株了解结核病进展的机制

基本信息

批准号：
10217964
负责人：
DAVID G RUSSELL
金额：
$ 64.79万
依托单位：
CORNELL UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-07-01 至 2022-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10217964
关键词：
Accounting Alveolar Alveolar Macrophages Awareness Bacteria Bronchoalveolar Lavage Cause of Death Cell surface Cells Cessation of life Collaborations Data Databases Dendritic Cells Development Disease Disease Progression Environment Goals Granuloma Growth HIV Health Heterogeneity Human Immune Immune response Immunologics Immunology Individual Infection Infection Control Infectious Agent Interferons Knowledge Literature Lung Macrophage Activation Malawi Mediating Methods Microbiology Modeling Monkeys Mus Mycobacterium tuberculosis Operative Surgical Procedures Pain Pathway interactions Penetrance Phagocytes Phenotype Physiological Population Program Development Protocols documentation Reporter Risk Route South Africa Suspensions System Testing Tuberculosis Tuberculosis Vaccines Uncertainty Universities Vaccines Validation bacterial fitness base cell type co-infection disorder control experimental study fitness flexibility human disease in vivo interstitial macrophage microbial mouse model mycobacterial neutrophil nonhuman primate novel permissiveness pulmonary granuloma recruit transcriptome sequencing volunteer

项目摘要

Project Summary / Abstract Due to its extensive penetrance of the human population, Mycobacterium tuberculosis (Mtb) remains a serious health risk to those individuals living with HIV. TB vaccine development programs are hampered by our poor understanding of the immune mechanisms underpinning disease progression. What we propose in this application is the utilization of Mtb reporter strains to provide a functional readout of microbial fitness and replication to enable us to identify and characterize those phagocytes that restrict bacterial growth (controllers) versus those phagocytes the promote bacterial growth (permissive) to understand the basis of disease progression in human tuberculosis. Our hypothesis is that Mtb reporter strains represent a novel route to the identification of the phagocyte populations that best restrict or promote bacterial replication, and that defining these cell populations will provide a rational framework for understanding immune control of tuberculosis. Aim 1: Development and validation of Mtb reporter strains and challenge models in the murine system. We will (a) exploit Mtb reporter strains to identify permissive and controller phagocyte populations; (b) optimize an ex vivo infection protocol for cells recruited to Mtb-infected mouse lung, to be used on NHP granulomas, and human airway and granuloma phagocytes; and (c) perform RNASeq profiling on the different phagocyte populations defined by the Mtb reporter strains for phenotypic analysis and to generate a panel of candidate cell surface markers for NHP and human ex vivo challenge studies. Aim 2. Functional characterization of phagocyte subsets in NHP pulmonary granulomas. We will perform parallel analysis of NHP phagocyte populations infected with reporter bacterial strains following the isolation of individual granulomas from infected monkey lungs. We propose comparable phenotypic characterization using biased and unbiased methods to functionally identify permissive and controller phagocyte populations in NHP tuberculosis in in vivo infections and ex vivo challenge experiments. Specific Aim 3: Determination of human phagocyte phenotypes through ex vivo challenge with Mtb reporter strains. In addition, in collaboration with Drs. Henry Mwandumba (Malawi) and Alasdair Leslie (South Africa) we will determine the functional phenotypes of human airway and human TB granuloma phagocyte subsets by probing the cells ex vivo with the fluorescent Mtb reporter strains.

项目概要/摘要由于其在人群中的广泛渗透性，结核分枝杆菌 (Mtb) 仍然是一种对艾滋病毒感染者造成严重的健康风险。结核病疫苗开发计划受到我们的阻碍对疾病进展的免疫机制了解甚少。我们在此提出的建议应用是利用 Mtb 报告菌株来提供微生物适应性的功能读数和复制使我们能够识别和表征那些限制细菌生长的吞噬细胞（控制器）与那些促进细菌生长（允许）的吞噬细胞相比，以了解疾病的基础人类结核病的进展。我们的假设是 Mtb 报告菌株代表了一种识别吞噬细胞的新途径最能限制或促进细菌复制的细胞群，并且定义这些细胞群将提供理解结核病免疫控制的合理框架。目标 1：在小鼠系统中开发和验证 Mtb 报告菌株和挑战模型。我们将 (a) 利用 Mtb 报告菌株来识别许可型和控制型吞噬细胞群； (b) 优化用于招募至 Mtb 感染小鼠肺部的细胞的离体感染方案，用于治疗 NHP 肉芽肿，以及人气道和肉芽肿吞噬细胞； (c) 对不同的吞噬细胞进行 RNASeq 分析由 Mtb 报告菌株定义的群体进行表型分析并生成一组候选细胞用于 NHP 和人类离体挑战研究的表面标记。目标 2. NHP 肺肉芽肿中吞噬细胞亚群的功能特征。我们将表演分离后感染报告菌株的 NHP 吞噬细胞群的平行分析来自受感染猴肺部的单个肉芽肿。我们提出使用可比较的表型表征有偏和无偏方法在功能上识别 NHP 中允许和控制吞噬细胞群体内感染和离体激发实验中的结核病。具体目标 3：通过 Mtb 离体攻击确定人类吞噬细胞表型记者菌株。此外，与博士合作。 Henry Mwandumba（马拉维）和 Alasdair Leslie（南非洲）我们将确定人类气道和人类结核肉芽肿吞噬细胞的功能表型通过用荧光 Mtb 报告菌株离体探测细胞来分析子集。