Identification of Mycobacterium tuberculosis-derived metabolites acting as ligands for MR1-restricted T cells.

鉴定作为 MR1 限制性 T 细胞配体的结核分枝杆菌衍生代谢物。

• 批准号: 10667695
• 负责人: Charles Kyriakos Vorkas
• 金额: $ 26.2万
• 依托单位: STATE UNIVERSITY NEW YORK STONY BROOK
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-04 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10667695
• 关键词: Anabolism; Antigen Presentation; Antigen-Presenting Cells; Antigens; Bacteria; Binding; Biological; Biological Assay; Blood; CRISPR interference; Candida; Cell physiology; Cells; Cellular biology; Chronic; Clinical; Clone Cells; Collaborations; Communities; Data; Disease; Event; Exposure to; Flow Cytometry; Future; Genetic; Genus Mycobacterium; Granzyme; Haitian; Household; Human; Immune; Immune Evasion; Immune Targeting; Immunity; Immunology; Immunotherapy; In Vitro; Infection; Interferons; Laboratories; Laboratory Study; Legionella; Ligands; Lymphocyte; Lymphocyte Biology; Lymphocyte Function; Measures; Metabolism; Microbe; Morbidity - disease rate; Mucous Membrane; Mus; Mycobacterium smegmatis; Mycobacterium tuberculosis; Patients; Peptides; Peripheral; Peripheral Blood Mononuclear Cell; Plasma; Preventive vaccine; Proteins; Research; Research Personnel; Riboflavin; Salmonella; Sampling; Source; System; T-Cell Activation; T-Cell Depletion; T-Cell Receptor; T-Lymphocyte; Technology; Tuberculosis; Vaccines; Vitamin B Complex; Work; cohort; cytotoxic; experience; improved; in vivo; insight; knock-down; liquid chromatography mass spectrometry; metabolome; metabolomics; microbial; mortality; mucosal vaccine; mutant; mycobacterial; novel; pathogen; protein purification; receptor; recruit; research clinical testing; response; transmission process

Project Summary Tuberculosis (TB) remains a leading global cause of morbidity and mortality, yet the immune correlates of protection remain elusive. Improved understanding of the initial immune events following Mycobacterium tuberculosis (Mtb) exposure that determine progression to infection and active disease is a critical priority for developing novel immune therapies and vaccines. Our laboratory studies innate lymphocyte biology and utilizes systems immunology approaches to identify targets for immune therapies against TB. In this proposal, we will focus on one of the most abundant innate lymphocytes responding to Mtb, mucosal-associated invariant T (MAIT) cells, that recognize activating/inhibitory microbial metabolites of riboflavin metabolism that act as non-peptide antigens presented by the evolutionarily conserved MHC I-related protein, MR1. While MAIT cells are activated by Mtb in vitro and in vivo, they do not robustly expand after infection. Moreover, MAIT cells are depleted during chronic murine and human TB disease. Our working hypothesis to explain this observation is that Mtb synthesizes an abundance of inhibitory MR1 ligands, which suppress MAIT cell activity and contribute to immune evasion. We have recently developed LC-MS technologies to detect MR1 ligands within biological samples and will couple this platform with our expertise in in mycobacterial genetics and TB immunology to define the MR1 metabolome of Mtb as compared to non-mycobacterial species as well as Mtb riboflavin auxotrophs we will generate by inducible CRISPRi knockdown to transiently perturb specific branchpoints of mycobacterial riboflavin biosynthesis. In parallel, we will quantify Mtb-specific MR1 ligands and MR1 expression/MAIT cell function in vivo during initial Mtb exposure and infection in humans who have been recently exposed to Mtb in their household. In sum, our approach will identify MR1 ligands specific to Mtb metabolism and determine their impact on MR1/MAIT cell activity, informing future targeting of Mtb Vitamin B metabolism in TB immune therapies and mucosal vaccines.

项目摘要 结核病（TB）仍然是全球发病率和死亡率的主要原因，但免疫系统 保护的相关性仍然难以捉摸。提高对初始免疫事件的理解 结核分枝杆菌（Mtb）暴露后，决定感染进展， 活动性疾病是开发新的免疫疗法和疫苗的关键优先事项。我们 实验室研究先天淋巴细胞生物学，并利用系统免疫学方法， 确定结核病免疫疗法的靶点。在本建议书中，我们将集中讨论其中一项 对Mtb，粘膜相关不变T（MAIT）应答的最丰富的先天性淋巴细胞 细胞，识别核黄素代谢的激活/抑制微生物代谢物， 由进化上保守的MHC I相关蛋白MR 1呈递的非肽抗原。 虽然MAIT细胞在体外和体内被Mtb激活，但它们在Mtb激活后不能稳健地扩增。 感染此外，MAIT细胞在慢性鼠和人TB疾病期间耗尽。我们 解释这一观察结果的工作假设是Mtb合成了大量的抑制性蛋白质， MR 1配体，其抑制MAIT细胞活性并有助于免疫逃避。我们有 最近开发了LC-MS技术，用于检测生物样品中的MR 1配体， 将这个平台与我们在分枝杆菌遗传学和结核病免疫学方面的专业知识结合起来， Mtb的MR 1代谢组与非分枝杆菌物种以及Mtb核黄素相比 我们将通过诱导型CRISPRi敲低产生营养缺陷型， 分枝杆菌核黄素生物合成的分支点。与此同时，我们将量化结核特异性 在初始Mtb暴露期间的体内MR 1配体和MR 1表达/MAIT细胞功能， 最近在家庭中接触过结核杆菌的人感染。总之，我们的 该方法将鉴定Mtb代谢特异性的MR 1配体，并确定它们对 MR 1/MAIT细胞活性，为结核免疫中Mt B维生素B代谢的未来靶向提供信息 治疗和粘膜疫苗。