Role of immune modulating butyrophilins in gamma delta T cell activation

免疫调节嗜丁蛋白在 γ δ T 细胞激活中的作用

• 批准号: 10118773
• 负责人: OLGA VINOGRADOVA
• 金额: $ 41.99万
• 依托单位: UNIVERSITY OF CONNECTICUT STORRS
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-27 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10118773
• 关键词: Address; Affect; Antigen-Presenting Cells; Antigens; Binding; Binding Proteins; Binding Sites; Biological; Biological Assay; Biological Models; Biology; Biophysics; CRISPR/Cas technology; Cell Line; Cell Surface Receptors; Cell physiology; Cell surface; Cells; Chemicals; Clinical Trials; Co-Immunoprecipitations; Communicable Diseases; Complex; Conflict (Psychology); Cryoelectron Microscopy; Cytolysis; Cytotoxic T-Lymphocytes; Detection; Disease; Distant; Drug Design; Epitopes; Exposure to; Extracellular Domain; Extracellular Protein; Family member; Fluorescence Resonance Energy Transfer; Genetic; Glycoproteins; Goals; Human; Immune; Immune response; Immunotherapy; In Vitro; Infection; Interferon Type I; K562 Cells; Lead; Length; Libraries; Ligand Binding; Ligands; Literature; Malignant Neoplasms; Mediating; Membrane; Modeling; Molecular; Molecular Conformation; Molecular Target; Mutate; Organizational Change; Peptides; Phosphorus; Point Mutation; Prodrugs; Production; Protein Isoforms; Proteins; Reporting; Research; Roentgen Rays; Role; Structural Models; Structure; Structure-Activity Relationship; T cell response; T-Cell Activation; T-Cell Receptor; T-Lymphocyte; Testing; Therapeutic; Therapeutic Agents; Visualization; Western Blotting; alpha-beta T-Cell Receptor; analytical ultracentrifugation; base; butyrophilin; cancer cell; clinical development; crosslink; cytokine; cytotoxic; dimer; disorder prevention; immune activation; immune checkpoint; insight; mutant; nanodisk; new therapeutic target; novel; novel therapeutics; preservation; protein complex; protein protein interaction; receptor; response; small molecule; structured data; therapy development; tool; γδ T cells

Abstract Immune cells protect us from disease by detecting and responding to foreign molecules. Understanding the molecular basis of this response is critical if we are to generate new therapies for treatment or prevention of diseases involving immune cells. The objective of our research is to characterize a newly discovered antigen detecting protein (butyrophilin 3, BTN3) which influences the immune response mediated by gamma delta T cells. Gamma delta T cells are cytotoxic T cells that respond quickly to foreign threats and serve a variety of roles, including direct lysis of infected or malignant cells, and as such, their activation holds great promise for therapeutic manipulation. In contrast to T cells that express the more prevalent alpha beta T cell receptor and respond to peptide antigens, T cells that express the Vgamma9Vdelta2 T cell receptor respond to small phosphorous-containing compounds known as phosphoantigens. Butyrophilin 3A1 (BTN3A1) is the receptor for phosphoantigens and mediates their activation of T cells through unclear mechanisms. We developed a library of novel synthetic phosphoantigens, as well as a library of butyrophilin constructs and point mutations, both of which are valuable tools for understanding the underlying biology of butyrophilins. Here, we propose aims that test the underlying hypothesis that ligand binding to the intracellular domain of BTN3A1 produces conformational and organizational changes that are required for interaction with counter receptors on T cells. Understanding how BTN3A1 and the related 3A2, 3A3, and 2A1 isoforms function at the molecular level is important because it 1) will help optimize past and present clinical trials that have examined phosphoantigens and phosphoantigen- expanded cells as immunotherapies, and 2) will identify new molecular targets or strategies within this complex for therapeutic manipulation. Our studies in Aim 1 will show a structural basis for how phosphoantigens affect the full length endogenous BTN3A1 using multiple biophysical and molecular biological approaches. In Aim 2, we will investigate the function of BTN3A1 in phosphoantigen-induced Vgamma9Vdelta2 T cell lysis of phosphoantigen containing cells and associated cytokine production. Together, this will allow us to build a structure-function model of BTN3 with regards to how its domain organization, oligomerization status, protein- protein interactions, and relationship to BTN2A1 influence its function. This will largely be done in the context of biological membranes through use of a novel in vitro membrane nanodisc/cryo-EM model system. Our ultimate goal is to present a clear structural model that demonstrates how phosphoantigen-induced conformational and/or compositional changes in the BTN3 complex promote effector functions of T cells. This will enable clinical development of therapies that modulate butyrophilin function to overcome immune checkpoints. These findings will come at a point when the biological understanding of antigen detection is far from complete, and thus have the potential to impact the field.

抽象的 免疫细胞通过检测和反应外国分子来保护我们免受疾病的影响。了解 如果我们要生成新的治疗或预防疗法，则这种反应的分子基础至关重要 涉及免疫细胞的疾病。我们研究的目的是表征新发现的抗原 检测蛋白质（丁二醇3，BTN3），它影响由伽马三角洲介导的免疫反应 细胞。伽马三角细胞是细胞毒性T细胞，对外国威胁的反应迅速并提供各种 角色，包括直接裂解感染或恶性细胞，因此，它们的激活对 治疗性操纵。与表达更为普遍的αβT细胞受体和的T细胞相反 对肽抗原的反应，表达vgamma9vdelta2 T细胞受体的T细胞对小 含磷酸化的化合物称为磷酸剂。丁二醇3A1（BTN3A1）是受体 磷酸剂并通过不清楚的机制介导其T细胞的激活。我们开发了一个图书馆 新型的合成磷酸剂，以及丁烷蛋白构建和点突变的库 这是理解丁二醇的潜在生物学的宝贵工具。在这里，我们提出的目的是 检验基本假设，即配体与BTN3A1的细胞内结构域结合产生构象 以及与T细胞上反对者相互作用所需的组织变化。理解 BTN3A1和相关的3A2、3A3和2A1同工型在分子水平上的功能很重要，因为它 1）将有助于优化过去和现在的临床试验，这些试验已经检查了磷酸剂和磷酸剂 - 扩展的细胞作为免疫疗法，2）将在该复合物中识别新的分子靶标或策略 用于治疗性操纵。我们在AIM 1中的研究将显示磷酸剂如何影响的结构基础 使用多种生物物理和分子生物学方法的全长内源性BTN3A1。在AIM 2中， 我们将研究BTN3A1在磷脂诱导的VGAMMA9VDELTA2 T细胞裂解中的功能 含有细胞的磷脂剂和相关的细胞因子产生。在一起，这将使我们能够建立一个 BTN3的结构 - 功能模型关于其域组织，寡聚状态，蛋白质的组织如何 蛋白质相互作用以及与BTN2A1的关系影响其功能。这将在很大程度上在 生物膜通过使用新型体外膜纳米盘/冷冻EM模型系统。我们的最终 目标是提出一个清晰的结构模型，该模型表明磷酸剂诱导的构象和/或 BTN3复合物的组成变化促进了T细胞的效应函数。这将启用临床 开发调节丁二醇功能以克服免疫检查点的疗法。这些发现 当对抗原检测的生物学理解远非完整，因此 影响该领域的潜力。