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

• 批准号: 10462661
• 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
• 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/10462661
• 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

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.

γδT细胞构成了针对感染剂的免疫激发和取消的重要组成部分 转化，但是它们通过其体形检测抗原的生化机制 重组T细胞受体（TCR）尚不清楚。与αβTCR不同，这些αβTCR仅限于识别抗原 在主要的组织相容性复合物（MHC）分子的背景下，γδTCR可以识别多种配体 从自我MHC到完整，未经加工的病毒糖蛋白。我们最近的工作已经建立了CD1 分子作为配体，用于对人Vδ1γδT细胞的亚群，产生鲁棒的功能，生化和 结构证据。我们试图将研究扩展到人类肠道，其中γδT细胞，尤其是Vδ1+ T细胞，占主导地位。我们的初步数据表明，CD1识别是强大的，并且在所有个体中都存在 检查了，并且肿瘤中的CD1反应性γδT细胞之间存在重要的功能差异 健康的毗邻组织。这是该提案的长期目标是充分表征此CD1反应性 肿瘤与健康组织中的种群，检查其功能效应子表型，TCR曲目和 除TCR外，免疫调节信号还塑造了募集，激活和潜在膨胀 这些细胞在高度相关的人类疾病（大肠癌）的背景下。我们的第一个目标， “在正常组织和解散组织中CD1特异性γδT细胞的表征。”，试图使用经典的细胞 通过直接的离体功能和转录分析完成的扩展，对CD1反应性和非 - 源自肿瘤和毗邻健康组织的反应性T细胞种群。这些数据将提供有关 与健康组织相比，调节肿瘤微环境中γδT细胞的信号。我们的第二个 目的，“阐明γδTCR与CD1/脂质复合物结合的分子机制。” 专注于表征这些细胞表达的γδTCR与CD1/脂质抗原之间的相互作用。 我们将使用蛋白质生物化学，生物物理学和X射线晶体学来阐明分子机制 γδTCR识别CD1/脂质。我们的努力将大大扩展我们对特定的理解 调节人类健康和疾病中γδT细胞活性的信号。我们的第三个目标：“确定存在 配体，共刺激性和/或共受体分子在CD1特异性γδT细胞激活中的作用和 结肠中的表型将表征可能调节的配体和免疫调节信号 在人类结直肠癌的背景下，CD1反应性γδT细胞的活性。我们将结合rnaseq，并 使用脐带血的分化测定，Vδ1细胞测试候选信号的相关性。这 通过OP9/DL1系统，将通过体外衍生的天然Vδ1T细胞完成。 γδT细胞可以是 促炎或调节性，因此，我们试图了解这些细胞在该疾病中扮演的角色（如果有的话） 状态。总之，这些目标将开始揭示人类免疫生物学中γδT细胞的奥秘 细胞和分子水平。