Novel Technologies for Investigation of Self-reactive T cells in MS and Relevant

研究多发性硬化症及相关疾病中自身反应性 T 细胞的新技术

• 批准号: 8583040
• 负责人: VIJAY K. KUCHROO
• 金额: $ 21.74万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-01 至 2018-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8583040
• 关键词: Adhesions; Animal Model; Antigen Presentation; Antigens; Area; Autoimmune Diseases; B-Lymphocytes; Baculoviruses; Behavior; Binding; C-terminal; CD28 gene; CD4 Positive T Lymphocytes; CD58 gene; CD80 gene; CLIP peptide; Cell Separation; Cell physiology; Cells; Chimeric Proteins; Clinical; Clonal Expansion; Collaborations; Communication; Communication Programs; Complex; Cytometry; Data Set; Dendritic Cells; Devices; Experimental Autoimmune Encephalomyelitis; Extracellular Domain; Funding; Gene Expression; Granulocyte-Macrophage Colony-Stimulating Factor; Histocompatibility Antigens Class II; Human; Image; Immune; Immunology; Individual; Inflammatory; Instruction; Intercellular adhesion molecule 1; Interleukin-17; Investigation; Label; Ligands; Link; Lipid Bilayers; Lipids; Love; Lymphatic Endothelial Cells; Lymphocyte; Measures; Micromanipulation; Mus; Myelin; Pathogenesis; Patients; Peptide/MHC Complex; Peptides; Phenotype; Production; Proteins; Reagent; Recombinant Proteins; Recombinants; Recovery; Regulatory T-Lymphocyte; Research Personnel; Resolution; Resources; Robotics; Role; Signal Transduction; Specificity; Specimen; Stromal Cells; Structure; Structure-Activity Relationship; Surface; Synapses; System; T-Cell Proliferation; T-Lymphocyte; Technology; Time; Tissues; Work; base; cell motility; chemokine; cytokine; functional group; genetic analysis; immunological synapse; immunological synapse formation; insight; interest; lymph nodes; mouse model; new technology; novel; podoplanin; programs; research study; response; single cell analysis; tool

This core will make important resources available to all Projects of this PPG. The nanowell technology represents a powerful tool to study the functional consequences of interactions between T cells and APC. The technology will enable in-depth investigation of the impact of different types of APCs on the functional programs of self-reactive T cells, which is of central importance to this PPG. A novel feature of this technology is that the functional consequences on both T cells and APC can be studied for interacting cell pairs, offering insights into the complex communication that programs self-reactive T cells with autoaggressive behavior. This core will enable integration of datasets from clinical specimens and experimental mouse models (Aim 1). It provides an important link between Projects 1 and 2, which both study myelin-specific T cells from patients with MS. The technology enables robotic isolation of cells of interest, such as myelin-specific T cells that produce combinations of pro-inflammatory cytokines (e.g. IL-17 + GM-CSF), for analysis of single-cell gene expression or clonal expansion. Furthermore, the technology will be made available for analysis of murine T cells, APCs and stromal cells (Projects 3 and 4). The core will also make recombinant proteins available to all Projects of this PPG, including experiments in the nanowell system (Aim 2), This core has provided such reagents to all Projects in the previous funding period and will continue to provide them for several efforts: 1. Studies by Dustin and Wucherpfennig have examined immunological synapse formation by human self-reactive T cells using the planar lipid bilayer system developed by the Dustin lab. Recent collaborative studies by the Love and Wucherpfennig labs have applied this approach to the nanowell system, enabling synapse structure to be related tp T cell function. These experiments require sets of human and murine recombinant proteins, including peptide-MHC complexes, ICAM-1 and CD80 (Projects 1 and 2). 2. Collaborative studies by the Kuchroo and Wucherpfennig labs have shown that tetramers of the MOG extracellular domain can be used to label MOG-specific B cells, and this reagent will be used in Project 3. 3. The turley, Kuchroo and Wucherpfennig labs have utilized Ig fusion proteins of podoplanin and CLEC-2 to study the function of these molecules in T cell - APC communication, which will be provided to Projects 3 and 4. This core will thus enable in-depth investigation of T cell - APC interactions in both humans and animal models to advance our understanding ofthe pathogenesis of MS.

该核心将为PPG的所有项目提供重要资源。纳米井技术