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CELL BIOLOGY AND MOLECULAR MECHANISMS OF HUMAN GAMMA/DELTA T CELL ACTIVATION

人类伽马/德尔塔 T 细胞激活的细胞生物学和分子机制

基本信息

批准号：
10166762
负责人：
Kaushik Choudhuri
金额：
$ 38.33万
依托单位：
UNIVERSITY OF MICHIGAN AT ANN ARBOR
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-06-11 至 2024-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10166762
关键词：
Address Affect Antigen Receptors Antigen Targeting Antigens Binding Biochemical Biological Biological Assay Biology Biophysics CRISPR/Cas technology Cell surface Cells Cellular Immunity Cellular Stress Cellular biology Cellular immunotherapy Chemicals Cytolysis Electron Microscopy Elements Epithelial Eukaryota Event Genetic Histocompatibility Human Image Immune Immunity Immunologic Surveillance Infection Interferon Type II Interleukin-17 Investigation Knowledge Leukemic Cell Lipids Lymphoblastic Leukemia Lymphocyte MHC antigen Malignant Neoplasms Mediating Medical Membrane Memory Metabolic Methods Microbe Modeling Molecular Mutagenesis Mutation Neutrophil Infiltration Pathway interactions Peptide/MHC Complex Peptides Phosphoric Monoester Hydrolases Phosphotransferases Population Predisposition Production Protein Family Proteins Public Health RNA Interference Research Resolution Role Signal Transduction Stress Structure Surface Synapses T cell response T-Cell Activation T-Lymphocyte Testing Therapeutic Virus ZAP-70 Gene adhesion receptor base butyrophilin cancer cell cancer type cytokine design effector T cell fluorescence imaging genome editing high resolution imaging imaging modality immune function immunological synapse improved innovation interdisciplinary approach mevalonate mucosal site novel pathogen peripheral blood receptor recruit response segregation spatiotemporal stoichiometry synaptic function tomography tumor γδ T cells

项目摘要

PROJECT SUMMARY/ABSTRACT Gamma/delta (γδ) T cells are innate-like lymphocytes that express unconventional antigen receptors (γδ TCR) on their surface. Unlike conventional αβ T cells, they do not recognize peptide-MHC antigens but are instead activated by invariant stress-inducible receptors and host/pathogen-associated metabolites. Most γδ T cells in human peripheral blood express the Vγ9Vδ2 TCR and are activated by phosphoisoprenoid (PiP) metabolites produced by medically important microbes, viruses and cancer cells. Vγ9Vδ2 T cell activation by PiPs is TCR- dependent, and requires contact with PiP-producing target cells that express the B7-family protein butyrophilin 3A1 (BTN3A1). However, the molecular mechanisms by which PiPs and BTN3A1 initiate signaling and activate Vγ9Vδ2 T cells remain unresolved. Our overall objective in this proposal is to identify the cell biological and molecular basis of PiP-dependent Vγ9Vδ2 T cell activation. In αβ T cells, antigen-induced signaling occurs at the cell-cell contact interface between T cells and antigen-bearing target cells, known as the immunological synapse. We propose to employ cutting edge approaches that include super-resolution fluorescence imaging, electron microscopy/tomography (EM), CRISPR/Cas9-based genomic editing, and proximity-based chemical tagging, to test our central hypothesis: that PiP-mediated Vγ9Vδ2 T cell activation occurs through a uniquely configured immunological synapse, in which TCR, accessory receptors (including BTN3A1), cytoskeletal elements, and lipid domains act in cis and across the synapse to initiate signaling. Our substantial preliminary investigations establish the feasibility of our approach and point to a need for `close contacts' and synaptic TCR/phosphatase segregation for effective PiP-dependent T cell activation. Surprisingly, we find that expression of BTN3A1 is required in both target cells and T cells, pointing to the possibility of an unsuspected trans-synaptic interaction of BTN3A1. To test our central hypothesis, we propose two Specific Aims: 1.) Using super-resolution imaging and EM, in conjunction with biochemical, signaling and functional assays, establish the PiP-dependent molecular organization, membrane topography, and key signaling events at the Vγ9Vδ2 T cell synapse; and 2.) Using genome-editing and RNAi approaches, combined with BTN3A1 structure/function mutagenesis, establish the role of PiP-induced changes in cell-surface organization of BTN3A1 in Vγ9Vδ2 T cell activation. Our approach is innovative as it uses cutting-edge imaging and biochemical methods to test a novel model for PiP-mediated Vγ9Vδ2T cell activation. The proposed research is significant as it will identify the fundamental biochemical, biophysical and cell biological requirements for Vγ9Vδ2 T cell activation, and provide a mechanistically-based framework to harness γδ T cell immunity for cellular immunotherapy.

项目总结/文摘