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Molecular and functional investigation of the role of CD1 in gamma delta T cell surveillance

CD1 在 γ δ T 细胞监测中作用的分子和功能研究

基本信息

批准号：
10268214
负责人：
Erin June Adams
金额：
$ 52.47万
依托单位：
UNIVERSITY OF CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-22 至 2025-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10268214
关键词：
Address Adoption Anti-Inflammatory Agents Antigens Autoimmunity Binding Biochemical Biochemistry Biological Assay Biophysics CD1 Antigens Cancerous Cell Culture Techniques Cell Line Cell physiology Cells Clonal Expansion Colon Colorectal Cancer Complement Complex Crystallization Data Disease Ensure Epithelial Foundations Frequencies Generations Glycoproteins Goals Health Human Immune response Immunity Immunobiology In Vitro Individual Infection Infectious Agent Inflammatory Institutional Review Boards Investigation Laboratories Lead Ligands Link Lipids Major Histocompatibility Complex Malignant Neoplasms Mediating Memory Molecular Mus Peripheral Phenotype Physiological Play Population Protein Biochemistry Protein Engineering RNA Role Sampling Shapes Signal Transduction Specificity Stress Structure Sulfoglycosphingolipids System T-Cell Activation T-Cell Receptor T-Lymphocyte T-cell receptor repertoire Testing Tissues Transcript Tumor-Derived Umbilical Cord Blood Viral Work X-Ray Crystallography adaptive immune response antigen detection biophysical analysis cell growth colorectal cancer progression cytokine functional status human disease immunoregulation innovation insight interest pathogen peripheral blood premalignant professor receptor receptor binding recruit response structural biology success tool transcriptome sequencing tumor tumor microenvironment tumorigenesis γδ T cells

项目摘要

γδ T cells constitute an important component of the immune response against infectious agents and cancerous transformations, yet the biochemical mechanisms by which they detect antigen through their somatically recombined T cell receptor (TCR) remain unclear. Unlike αβTCRs, which are restricted to recognizing antigens in the context of Major Histocompatibility Complex (MHC) molecules, γδTCRs can recognize a diversity of ligands ranging from self MHC to intact, unprocessed, viral glycoproteins. Our recent work has established CD1 molecules as ligands for a subpopulation of human Vδ1 γδ T cells, producing robust functional, biochemical and structural evidence. We seek to extend our studies to the human gut, where γδ T cells, and in particular, Vδ1+ T cells, predominate. Our preliminary data suggests that CD1 recognition is robust and present in all individuals examined, and that there exist important functional differences between CD1 reactive γδ T cells in tumors versus healthy adjoining tissue. Thus, the long-term goal of this proposal is to fully characterize this CD1 reactive population in tumors versus healthy tissue, examining their functional effector phenotypes, TCR repertoire and immunomodulatory signals, in addition to the TCR, that shape the recruitment, activation and potential expansion of these cells in the context of a highly relevant human disease, colorectal cancer. Our first aim, “Characterization of CD1-specific γδ T cells in normal and diseased tissue.”, seeks to use classical cellular expansions complemented by direct ex vivo functional and transcript analysis to profile CD1 reactive and non- reactive T cell populations derived from tumor and adjoining healthy tissue. These data will provide insight into the signals that regulate γδ T cells within the tumor microenvironment compared to healthy tissue. Our second aim, “Elucidation of the molecular mechanisms by which γδ TCRs bind to CD1/lipid complexes.”, will focus on characterizing the interaction between the γδ TCRs expressed by these cells and CD1/lipid antigen. We will use protein biochemistry, biophysics and x-ray crystallography to elucide the molecular mechanisms by which the γδ TCR recognizes CD1/lipid. Our effort will significantly expand our understanding of the specific signals that regulate γδ T cell activity in human health and disease. Our third aim, “Determine the presence and role of ligand, co-stimulatory and/or co-receptor molecules in CD1 specific γδ T cell activation and phenotype in the colon” will characterize the ligand and immunomodulatory signals that may regulate the activity of CD1 reactive γδ T cells in the context of human colorectal cancer. We will combine RNAseq and differentiation assays using cord blood derived, naïve Vδ1 cells to test the relevance of candidate signals. This will be complemented by in vitro derived native Vδ1 T cells through the OP9/DL1 system. γδ T cells can be either pro-inflammatory or regulatory, therefore we seek to understand which role these cells play, if any, in this disease state. Together, these aims will begin to unravel the mystery of γδ T cells in human immunobiology, both at the cellular and molecular levels.

γδ T 细胞是针对传染性病原体和癌症的免疫反应的重要组成部分。转化，但它们通过体细胞检测抗原的生化机制重组 T 细胞受体 (TCR) 仍不清楚。与 αβTCR 不同，αβTCR 仅限于识别抗原在主要组织相容性复合物 (MHC) 分子的背景下，γδTCR 可以识别多种配体范围从自身 MHC 到完整、未加工的病毒糖蛋白。我们最近的工作建立了CD1 分子作为人类 Vδ1 γδ T 细胞亚群的配体，产生强大的功能、生化和结构性证据。我们寻求将我们的研究扩展到人类肠道，其中 γδ T 细胞，特别是 Vδ1+ T细胞占主导地位。我们的初步数据表明 CD1 识别能力很强并且存在于所有个体中研究发现，肿瘤中的 CD1 反应性 γδ T 细胞与肿瘤中的 CD1 反应性 γδ T 细胞之间存在重要的功能差异。健康的邻近组织。因此，该提案的长期目标是充分表征这种 CD1 反应性肿瘤与健康组织中的群体，检查它们的功能效应表型、TCR 库和除了 TCR 之外，免疫调节信号还影响招募、激活和潜在扩展在高度相关的人类疾病——结直肠癌的背景下研究这些细胞。我们的首要目标， “正常和患病组织中 CD1 特异性 γδ T 细胞的表征。”，寻求使用经典的细胞通过直接离体功能和转录本分析来补充扩展，以分析 CD1 反应性和非反应性来自肿瘤和邻近健康组织的反应性 T 细胞群。这些数据将提供深入了解与健康组织相比，调节肿瘤微环境中 γδ T 细胞的信号。我们的第二个目标是“阐明 γδ TCR 与 CD1/脂质复合物结合的分子机制”。重点关注这些细胞表达的 γδ TCR 与 CD1/脂质抗原之间的相互作用。我们将利用蛋白质生物化学、生物物理学和 X 射线晶体学来阐明分子机制 γδ TCR 识别 CD1/脂质。我们的努力将显着扩大我们对特定领域的理解调节人类健康和疾病中 γδ T 细胞活性的信号。我们的第三个目标是“确定存在配体、共刺激和/或共受体分子在 CD1 特异性 γδ T 细胞激活中的作用和结肠中的表型”将表征可能调节结肠的配体和免疫调节信号 CD1 反应性 γδ T 细胞在人类结直肠癌中的活性。我们将结合 RNAseq 和使用脐带血来源的初始 Vδ1 细胞进行分化测定，以测试候选信号的相关性。这将通过 OP9/DL1 系统得到体外衍生的天然 Vδ1 T 细胞的补充。 γδ T 细胞可以是促炎性或调节性，因此我们试图了解这些细胞在这种疾病中发挥什么作用（如果有的话）状态。总之，这些目标将开始揭开 γδ T 细胞在人类免疫生物学中的神秘面纱。细胞和分子水平。