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) 影响着全球约 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，导致其代谢能力显着下降。产生胰岛素，浸润胰岛并介导  细胞，从而产生糖尿病性 CD8+ 和 CD4+ 效应 T 细胞通过识别 I 类和 II 类 MHC 上呈现的肽表位，以抗原特异性方式进行破坏相反，调节性 T 细胞可以抑制致糖尿病 T 细胞，从而预防 T1D。识别  细胞呈递的表位对于这些自身反应性 T 的功能至关重要。仅少数自身表位被自身反应性 CD8+、CD4+ 效应子和调节性 T 细胞识别。然而，T1D 细胞中的表位已被大多数胰岛浸润性 T 细胞识别。这些表位的知识对于了解疾病发病机制和了解至关重要。目前，广泛适用且有效的 T 细胞抗原发现方法。该项目的总体目标是揭示自身反应性。 T1D 中的自身反应性 T 细胞使用我们开发的新颖且通用的抗原发现技术在本提案中，我们将利用信号传导和抗原呈递来发现 T 细胞表位。双功能受体 (SABR) 用于识别小鼠中效应 T 细胞和调节 T 细胞识别的表位我们建议构建源自表达基因的表位文库。特别是在胰腺  细胞中，我们将鉴定出胰岛浸润 T 细胞的新靶点。根据已发表的 NOD 小鼠质谱和基因表达数据集构建表位库。我们将通过对 NOD 小鼠的胰岛进行单细胞 TCR 测序来获得胰岛反应性 TCR。使用  细胞衍生的 SABR 文库，我们将确定胰岛反应性 TCR 的同源表位并验证这些研究确定的表位将导致未来的研究旨在了解这些表位。自身反应性 T 细胞破坏免疫耐受并开发靶向免疫治疗方法这种方法也将为从 T1D 患者中发现 T 细胞的抗原奠定基础。样本隶属于人类胰岛研究网络。 1