Gamma Delta T cell surveillance of metabolites as signals of cellular stress

Gamma Delta T 细胞对代谢物的监测作为细胞应激信号

基本信息

批准号：
9171943
负责人：
Erin June Adams
金额：
$ 38.94万
依托单位：
UNIVERSITY OF CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-11-03 至 2019-10-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9171943
关键词：
Actins Adopted Agonist Alpha Cell Anabolism Antibodies Antigens Applications Grants Binding Blood Cell membrane Cell surface Cells Cellular Stress Clinical Complement Coupled Crystallography Cytoskeleton Data Detection Disease Energy Transfer Event Extracellular Domain Fluorescence Goals Health Human Immobilization Immunity Individual Infection Laboratories Lead Leprosy Ligands Link Maps Mass Spectrum Analysis Measurement Mediating Mediator of activation protein Membrane Microbe Microscopy Modeling Molecular Molecular Conformation Mycobacterium leprae Mycobacterium tuberculosis Pathway interactions Play Population Primates Process Production Protein Engineering Protein Family Proteins Publications RNA interference screen Research Resolution Role Signal Transduction Sirolimus Structure Surface T cell response T-Cell Activation T-Cell Receptor T-Lymphocyte T-Lymphocyte Subsets Testing Translating Tuberculosis antigen binding base butyrophilin crosslink experimental study extracellular functional mimics genome-wide human disease improved in vivo isoprenoid knock-down member mevalonate microbial neoplastic cell pathogen public health relevance response sensor γδ T cells

项目摘要

DESCRIPTION (provided by applicant): Human V9V2 T cells are the major subset of  T cells that are found in human blood, comprising up to 5% of the T cells in healthy individuals and expanding to 20-50% during infection or disease. These cells play important roles in mediating immunity against microbial pathogens, including Mycobacterium tuberculosis and Mycobacterium leprae (the causative agents of tuberculosis and leprosy, respectively), and can respond potently against certain types of tumor cells. V9V2 T cells respond to these threats through recognition of structurally related non-peptidic phosphorylated antigens (pAgs). In tumor cells these are intermediate metabolites that accumulate due to over-production of the mevalonate pathway and in microbes they are generated during isoprenoid biosynthesis. It is largely unknown how V9V2 T cells recognize these pAgs; this proposal seeks to improve our overall understanding of the mechanisms behind V9V2 T cell activation and improve our ability to modulate this population in clinical settings. These cells also represent a prime opportunity to study alternative recognition strategies by  T cells, as classical and most no-classical MHC molecules do not appear to be involved in the recognition process. Our collaborators were the first to define the role of the BTN3A molecules in pAg-mediated V9V2 T cell stimulation and we have since then demonstrated the intracellular domain of one on them, BTN3A1, is the molecular sensor for pAg accumulation. We seek to understand the events that occur after pAg binding in our Aim1: "To determine the direct molecular consequences of pAg binding to the BTN3A1 B30.2 intracellular domain." by using structural and dynamic studies including crystallography, NMR and fluorescence based measurements (FRET). In our Aim 2. "To determine the supra-molecular organization of BTN3A1 molecules in the plasma membrane upon pAg or 20.1 mAb binding and its role in V9V2 T cell activation", we will use FRET based approaches, microscopy and protein engineering to study the factors that mediate cell surface assembly of BTN3A molecules. Finally, we will employ cross-linking and pull-downs combined with a SILAC-based mass spectrometry approach, complemented with a high-throughput genome-wide knockdown screen to pursue our Aim 3. "From BTN3A1 to the V9V2 TCR: discovery of the relevant molecular players linking BTN3A1 to V9V2 T cell activation", which will define the other molecular players that are involved in pAg recognition in inducing the signals that directly lead to V9V2 T cell activation.

描述（由适用提供）：人V9V2T细胞是人类血液中发现的T细胞的主要子集，在健康个体中最多可完成5％的T细胞，并在感染或疾病期间膨胀到20-50％。这些细胞在介导免疫原中的重要作用，包括结核分枝杆菌和麻风分枝杆菌（分别是结核病和麻风病的病因），并且可以对某些类型的肿瘤细胞反应。 V9V2T细胞通过识别结构相关的非肽磷酸化抗原（PAG）来应对这些威胁。在肿瘤细胞中，这些是中间代谢产物，由于甲丙酸酯途径的过量产生而积累，在类异急动物生物合成期间产生的微生物。 V9V2T细胞在很大程度上尚不清楚这些PAG。该提案旨在提高我们对V9V2T细胞激活背后机制的总体理解，并提高我们在临床环境中调节该人群的能力。这些细胞也代表了研究T细胞的替代识别策略的主要机会，因为古典和大多数不经典的MHC分子似乎与识别过程无关。我们的合作者是第一个定义BTN3A分子在PAG介导的V9V2T细胞刺激中的作用的人，此后我们已经证明了其中一个上的细胞内结构域BTN3A1，BTN3A1是PAG积累的分子传感器。我们试图了解我们的AIM1中PAG结合后发生的事件：“确定PAG与BTN3A1 B30.2细胞内结构域的直接分子后果。”通过使用结构和动态研究，包括晶体学，NMR和基于荧光的测量（FRET）。在我们的目标2中。“为了确定PAG时质膜中BTN3A1分子的超分子组织，或20.1 mAb结合及其在V9V2T细胞激活中的作用及其在V9V2T细胞激活中的作用”，我们将使用基于FRET的方法，显微镜和蛋白质工程来研究介导Btn3a Melecules的细胞表面组装的因素。 Finally, we will employ cross-linking and pull-downs combined with a SILAC-based mass spectrometry approach, completed with a high-throughput genome-wide knockdown screen to pursue our Aim 3. "From BTN3A1 to the V9V2 TCR: discovery of the relevant molecular players linking BTN3A1 to V9V2 T cell activation", which will define the other molecular players that are involved in pAg识别在诱导的信号中直接导致V9V2T细胞激活。