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; 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）仍然是发病率和死亡率的全球主要原因，但免疫 保护的关联仍然难以捉摸。改善了对最初免疫事件的理解 跟随结核分枝杆菌（MTB）暴露，确定了感染的发展和 活动疾病是开发新型免疫疗法和疫苗的关键优先事项。我们的 实验室研究先天淋巴细胞生物学，并利用系统免疫学方法 确定针对结核病的免疫疗法的靶标。在此提案中，我们将专注于 对MTB，与粘膜相关的不变t（MAIT）响应MTB的最丰富的先天淋巴细胞（MAIT） 识别核黄素代谢的激活/抑制性微生物代谢产物的细胞 由进化保守的MHC I相关蛋白MR1提出的非肽抗原。 虽然Mait细胞在体外和体内被MTB激活，但它们在 感染。此外，在慢性鼠和人类结核病疾病期间，MAIT细胞会耗尽。我们的 解释这一观察的工作假设是MTB综合了丰富的抑制 MR1配体，抑制Mait细胞活性并导致免疫逃避。我们有 最近开发了LC-MS技术来检测生物样品中的MR1配体，并将 将这个平台与分枝杆菌遗传学和结核病免疫学方面的专业知识相结合，以定义 MR1代谢组MTB与非细菌物种以及MTB核黄素相比 我们将通过诱导的CRISPRI敲低以瞬时扰动特异性产生的合并营养 分枝杆菌核黄素生物合成的分支。同时，我们将量化MTB特异性 在初始MTB暴露期间，MR1配体和MR1表达/MAIT细胞功能在体内和 最近在其家庭中接触MTB的人类感染。总而言之 方法将识别特定于MTB代谢的MR1配体并确定其对 MR1/MAIT细胞活性，告知MTB维生素B代谢的未来靶向结核病免疫 疗法和粘膜疫苗。