The Origins of Human Anti-Insulin B Lymphocytes in Type 1 Diabetes

1 型糖尿病中人类抗胰岛素 B 淋巴细胞的起源

基本信息

批准号：
10343084
负责人：
Rachel H Bonami
金额：
$ 45.43万
依托单位：
VANDERBILT UNIVERSITY MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-12-01 至 2026-11-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10343084
关键词：
Address Affinity Antigen-Presenting Cells Antigens Autoantibodies Autoantigens Autoimmune Autoimmune Diseases Autoimmune Responses Autoimmunity Avidity B cell clonality B cell differentiation B-Cell Antigen Receptor B-Cell Receptor Binding B-Lymphocyte Subsets B-Lymphocytes B-cell receptor repertoire sequencing Beta Cell Binding Biological Assay Biological Markers Blood Blood Volume Cell Death Cell Differentiation process Cells Cellular Indexing of Transcriptomes and Epitopes by Sequencing Child Clinical Clinical Trials Clonal Evolution Clonal Expansion Coal Code Dangerousness Data Detection Diabetes Mellitus Disease Disease Progression Epitope Mapping Epitopes Event Future Goals Heterogeneity Human Hybridomas IA-2 protein Immune Targeting Immune Tolerance Immune response Immune signaling Immunoglobulin Genes Immunoglobulin-Secreting Cells Immunoglobulins Individual Insulin Insulin-Dependent Diabetes Mellitus Intervention Investigation Investigational Therapies Islets of Langerhans Knowledge Lymphoma Maps Measures Memory Memory B-Lymphocyte Monitor Mus Mutate Mutation Participant Pathogenicity Pathologic Pathway interactions Patients Phenotype Population Process Production Recombinants Risk Role Sampling Sequence Analysis Serinus Shapes Signal Transduction Specificity Structure of germinal center of lymph node T-Lymphocyte Techniques Technology Testing Therapeutic Time Visit analysis pipeline antigen binding autoreactive B cell autoreactivity biobank defined contribution experimental study glucose tolerance human monoclonal antibodies immunological intervention impaired glucose tolerance inclusion criteria islet islet cell antibody molecular sequence database peripheral blood predictive marker prevent relapse prediction research clinical testing response screening single cell analysis

项目摘要

PROJECT SUMMARY B lymphocytes orchestrate autoimmune beta cell attack in type 1 diabetes (T1D) by presenting autoantigen to T cells which kill beta cells. Circulating insulin autoantibodies (IAAs) are not directly pathogenic but help predict T1D by signaling this dangerous B/T lymphocyte crosstalk. Immune targeting of insulin-producing beta cells occurs for months or even decades before symptomatic diabetes onset. Heterogeneity in diabetes progression and clinical trial responses slow the search for a T1D cure. Better biomarkers to identify and mechanistically explain responders are needed to overcome this bottleneck. Protective immune responses typically arise by T cell selection and affinity maturation of B lymphocytes in germinal centers, resulting in memory B lymphocyte and antibody-secreting cell differentiation. However, autoimmune responses do not always follow this pathway. For example, we find anti-insulin B cells (AIBCs) accelerate diabetes in T1D-prone mice, yet few AIBCs differentiate into IAA-secreting cells, despite entering germinal centers. To fill gaps in knowledge AIBC expansion early in T1D, we built a unique, carefully-curated biobank of pre-symptomatic T1D TrialNet participants. We find AIBCs in the peripheral blood of IAA-negative pre-symptomatic T1D donors, demonstrating B lymphocyte autoimmunity for insulin evades conventional detection via circulating IAAs in a subset of at-risk individuals. Longitudinal sampling and clinical testing at each T1D TrialNet visit provide opportunities to identify donor-specific changes as T1D progresses from stage 1 (normal glucose tolerance) to stage 2 (impaired glucose tolerance) and stage 3 (diabetes). We find AIBCs are skewed towards a memory B cell phenotype. Clonally-expanded, memory B lymphocytes express germline and minimally-mutated B cell receptors in stage 1 T1D donors. These data support our hypothesis that AIBCs expand and enter the memory compartment prior to glucose tolerance impairment, sometimes without IAA production. We will integrate AIBC phenotype, B cell receptor (immunoglobulin) sequence identity, clonal relatedness, B cell receptor affinity/avidity for insulin, and insulin epitope mapping to identify features that govern insulin recognition. High- throughput single cell analysis of B cell receptor repertoire paired with cell phenotype will identify changes in the same donor over time and with T1D stage progression to determine B lymphocyte repertoire and subset shifts that occur as glucose tolerance is lost. In addition to memory skewing, AIBCs show biased V and J immunoglobulin gene use. We will track this combination of features and use LIBRAseq to identify candidate autoreactive BCRs to recombinantly express and test for islet autoantigen recognition, including other known islet autoantigens (GAD65, IA-2, ICA512, and ZNT8). The human monoclonal antibodies, anti-insulin BCR sequence database, and analysis pipeline/code we will develop will be made publicly available. These studies are a necessary step towards using AIBCs as biomarkers in clinical trials to identify favorable changes in cellular autoimmunity and zero in on specific changes AIBCs undergo to deconvolute response heterogeneity.

项目摘要 B淋巴细胞通过将自动抗原呈现为1型糖尿病（T1D）的自动免疫性β细胞攻击杀死β细胞的T细胞。循环胰岛素自身抗体（IAAS）不是直接的致病性，但有助于预测 T1D通过向这种危险的B/T淋巴细胞串扰发出信号。免疫靶向产生胰岛素的β细胞发生在有症状的糖尿病发作之前数月甚至几十年。糖尿病进展的异质性和临床试验反应减慢了对T1D治疗的搜索。更好地识别和机械识别和机械的生物标志物需要说明响应者以克服这种瓶颈。保护性免疫反应通常是由T B淋巴细胞的细胞选择和亲和力成熟在生发中心，导致记忆B淋巴细胞和分泌细胞分化的抗体。但是，自身免疫反应并不总是遵循此途径。例如，我们发现抗胰岛素B细胞（AIBC）在T1D-易于发挥的小鼠中加速糖尿病，但很少有AIBCS 尽管进入生发中心，但分化为分泌IAA的细胞。填补知识的空白AIBC 在T1D的早期扩展，我们建立了一个独特的，精心策划的生物库，该生物库是症状前T1D试验网参与者。我们在IAA阴性症状前T1D供体的外周血中发现AIBC，证明B淋巴细胞自身免疫性胰岛素通过循环IaA逃避常规检测处于危险的人的子集。每次T1D试验访问的纵向采样和临床测试提供随着T1D从第1阶段（正常葡萄糖耐受性）发展到第2阶段（葡萄糖耐受性受损）和第3期（糖尿病）。我们发现AIBC偏向记忆b 细胞表型。克隆扩张，记忆B淋巴细胞表达种系和最小氧化的B细胞第1阶段T1D供体中的受体。这些数据支持我们AIBC扩展并输入内存的假设葡萄糖耐量损害之前的隔室，有时没有IAA产生。我们将整合AIBC 表型，B细胞受体（免疫球蛋白）序列身份，克隆相关性，B细胞受体胰岛素的亲和力/亲和力/亲和力和胰岛素表位映射以识别控制胰岛素识别的特征。高的- 与细胞表型配对的B细胞受体库的吞吐量单细胞分析将确定变化随着时间的推移和T1D阶段的进展，相同的供体以确定B淋巴细胞库和子集随着葡萄糖耐受性而发生的变化。除了记忆偏斜之外，AIBC还显示出偏见的V和J 免疫球蛋白基因的使用。我们将跟踪这种功能的组合，并使用Libraseq识别候选人自动反应性BCR重组表达和测试胰岛自动抗原识别，包括其他已知的胰岛自动抗原（GAD65，IA-2，ICA512和ZNT8）。人类单克隆抗体，抗胰岛素BCR 我们将开发的序列数据库和分析管道/代码将公开可用。这些研究是在临床试验中使用AIBC作为生物标志物的必要步骤，以确定有利的变化细胞自身免疫性和零对特定变化的AIBC经历了反应异质性。