Identification of the cognate epitopes of autoreactive T cells in Type 1 Diabetes

1 型糖尿病自身反应性 T 细胞同源表位的鉴定

基本信息

批准号：
10264075
负责人：
Alok joglekar
金额：
$ 15.91万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
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：
2020-09-14 至 2022-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10264075
关键词：
Affect Alleles American Antigens Autoantigens Autoimmune Binding Biological Assay CD4 Antigens CD4 Positive T Lymphocytes CD8B1 gene Cells Clone Cells Data Set Disease Epitopes FOXP3 gene Foundations Future Gene Expression Genes Goals Histocompatibility Antigens Class II Human Immune Tolerance Immune response Immune system Immunotherapy In Vitro Inbred NOD Mice Individual Insulin Insulin-Dependent Diabetes Mellitus Islets of Langerhans Jurkat Cells Knowledge Libraries Major Histocompatibility Complex Maps Mass Spectrum Analysis Mediating Methods Mus Pancreas Pathogenesis Patients Peptide/MHC Complex Peptides Protocols documentation Publishing Regulatory T-Lymphocyte Reporter Research Sampling Signal Transduction Sorting - Cell Movement Source Specificity T cell receptor repertoire sequencing T-Cell Antigen Receptor Specificity T-Cell Receptor T-Lymphocyte T-Lymphocyte Epitopes T-cell receptor repertoire Techniques Technology Tissues arm autoreactive T cell autoreactivity base cell killing central tolerance combat cytotoxicity design diabetes pathogenesis diabetogenic effector T cell in vitro Assay islet mouse model novel novel therapeutic intervention peripheral tolerance prevent programs receptor single cell sequencing targeted treatment

项目摘要

ABSTRACT Type 1 Diabetes (T1D), affects approximately 4 million individuals worldwide, including 1.6 million Americans. T1D is caused by progressive destruction of pancreatic  cells, resulting in a significantly diminished capacity to produce insulin. Diabetogenic CD8+ and CD4+ effector T cells that infiltrate pancreatic islets mediate  cell destruction in an antigen-specific manner by recognizing peptide epitopes presented on class I and class II MHC molecules respectively. In contrast, regulatory T cells can suppress diabetogenic T cells, thereby preventing T1D pathogenesis. Recognition of epitopes presented by  cells is critical for the function of these autoreactive T cells. Only a small number of self-epitopes recognized by autoreactive CD8+, CD4+ effector and regulatory T cells in T1D have been uncovered. However, the epitopes recognized by the majority of islet-infiltrating T cells are not known. The knowledge of these epitopes is critical for understanding disease pathogenesis and for developing targeted therapies. Currently, widely applicable and efficient methods for T cell antigen discovery for uncovering autoreactivity are lacking. The overarching goal of this project is to uncover the cognate epitopes of autoreactive T cells in T1D using a novel and generalizable antigen discovery technology developed by our group. In this proposal, we will employ T cell epitope discovery using Signaling and Antigen-presenting Bifunctional Receptors (SABRs) to identify the epitopes recognized by effector and regulatory T cells in a mouse model of T1D, NOD mice. We propose that constructing a library of epitopes derived from genes expressed specifically in pancreatic  cells will lead to identification of novel targets of islet-infiltrating T cells. We will construct epitope libraries from published mass spectrometry and gene expression datasets from NOD mice. We will obtain islet-reactive TCRs by performing single cell TCR sequencing on pancreatic islets of NOD mice. Using  cell derived SABR libraries, we will determine the cognate epitopes of islet-reactive TCRs and validate them in vitro. The epitopes identified by these studies will lead to future studies aiming to understand the breakage of immune tolerance by autoreactive T cells and to develop targeted immunotherapy approaches to combat T1D. This approach will also establish the foundation for antigen discovery for T cells from T1D patient samples in affiliation with Human Islet Research Network. 1

抽象的 1型糖尿病（T1D）影响了全球约400万人，其中包括160万美国人。 T1D是由胰腺细胞进行性破坏引起的，导致能力显着降低产生的胰岛素。浸润胰岛的糖尿病性CD8+和CD4+效应T细胞介导细胞通过识别在I类和II类MHC上呈现的肽表位，以抗原特异性方式破坏分子分别。相反，调节性T细胞可以抑制糖尿病性T细胞，从而防止T1D 发病。 细胞提出的表位的识别对于这些自动反应t的功能至关重要细胞。仅自动反应性CD8+，CD4+效应子和调节t识别的少数自我介绍 T1D中的细胞已被发现。但是，大多数胰岛浸润的T细胞所识别的表位不知道。这些表位的知识对于理解疾病发病机理和开发目标疗法。当前，广泛适用和有效的T细胞抗原发现方法缺乏发现自动反应性。该项目的总体目标是发现 T1D中的自动反应性T细胞使用我们的新颖且可推广的抗原发现技术团体。在此提案中，我们将使用信号传导和抗原呈现T细胞表位发现双功能受体（SABR）识别小鼠效应子和调节性T细胞识别的表位 T1D的模型，点头小鼠。我们建议构建从表达的基因得出的表位库特别是在胰腺细胞中，将导致识别胰岛渗透T细胞的新靶标。我们将从已发表的质谱和基因表达数据集中构造表位库。我们将通过在NOD小鼠的胰岛上进行单细胞TCR测序来获得胰岛反应性TCR。使用细胞得出的SABR库，我们将确定胰岛反应性TCR的同源表位并验证他们体外。这些研究确定的表位将导致未来的研究旨在了解自身反应性T细胞破坏免疫耐受性，并开发出靶向免疫疗法的方法战斗T1D。这种方法还将为T1D患者的T细胞发现抗原发现基础与人类胰岛研究网络的相关样品。 1