AUTOANTIBODY ANALYSIS FOR A BETTER PREDICTION OF TYPE 1 DIABETES

自身抗体分析可更好地预测 1 型糖尿病

• 批准号: 7468455
• 负责人: AKE LERNMARK
• 金额: $ 19.11万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-07-01 至 2008-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7468455
• 关键词: Accounting; Affinity; Age; Alleles; Antibodies; Antibody Affinity; Antibody Avidity; Antigen Targeting; Antigens; Autoantibodies; Autoantigens; Autoimmunity; Avidity; B-Lymphocytes; Binding; Binding Sites; Biological Assay; Blood specimen; CD4 Positive T Lymphocytes; Cell Line; Characteristics; Chromosomes, Human, Pair 11; Complementary DNA; Data; Dependency; Diabetes Mellitus; Diabetes autoantibodies; Diagnostic Sensitivity; Disease; Disease Progression; Disease model; Epitopes; Fab Immunoglobulins; General Population; Genetic; Genetic Predisposition to Disease; Genotype; Glutamate Decarboxylase; Hybridomas; Hybrids; IgE; Immunoglobulin A; Immunoglobulin G; Immunoglobulin M; Individual; Insulin; Insulin-Dependent Diabetes Mellitus; Islet Cell; Islets of Langerhans; Ligands; Measures; Minisatellite Repeats; Molecular Conformation; Monoclonal Antibodies; Pathogenesis; Patients; Pattern; Phase; Phenotype; Populations at Risk; Process; Protein Isoforms; Protein Tyrosine Phosphatase; Proteins; Recombinants; Regulation; Reporting; Resistance; Risk; Risk Factors; Role; Sequence Analysis; Serum; Specificity; Sum; T-Cell Proliferation; T-Lymphocyte; Testing; To autoantigen; cohort; cytokine; diabetes risk; hybrid protein; islet; pre-clinical; programs; research study; response

Type 1 diabetes (T1D) is predicted by autoantibodies (Ab) directed against the 65kD isoform of glutamic acid decarboxylase (GAD65), IA-2, -a protein tyrosine phosphatase-like molecule-, and insulin. The objective is to uncover the mechanisms by which GAD65, IA-2, and insulin autoantibodies predict T1D. The overall hypothesis is that both antibody avidity and epitope specificity mature as an individual progresses towards TED reflecting antigen-driven processes. The aims are: 1. To test the hypothesis that subtype and isotype autoantibodies to GAD65, IA-2, and insulin, alone or in combination predict progression from islet autoimmunity to T1D. Specific subtype and isotype autoantibodies against GAD65, IA-2, and insulin will be analyzed as a function of progression or non-progression to T1D. 2. To test the hypothesis that progression to T1D is associated with autoantibody maturation against specific autoantigen epitopes. Using isoform and subtype specific assays with both GAD65/67 hybrid proteins and competing epitope-specific recombinant Fab, autoantibody maturation to specific epitopes will be established as a function of progression or non-progression to T1D. 3. To test the hypothesis that progression to T1D is associated with the avidity of epitope-specific autoantibodies. Titers and autoantibody affinity maturation will be examined in competition assays with recombinant Fab to unique epitopes of GAD65 and insulin. Measures of autoantibody avidity and affinity constants will be used to distinguish non-progressors from progressors to T1D. Project 2 will collaborate with a) Project 1 on non-progressors, progressors and new onset T1D patients and CD4 T cell profiling in relation to the autoantibody characteristics; b) project 3 on new onset classic T1D patients compared to type 1.5 diabetes subjects and T cell proliferation to multiple islet cell proteins and cytokine secretion patterns and c) Project 4 HLA and non-HLA genetic factors in general population subjects of T1D risk followed until islet autoimmunity and T1D. Core B will be instrumental to sequence Fab cDNA cloned from hybridoma cell lines producing monoclonal antibodies to GAD65, IA2, and insulin. We expect to find that autoantibody maturation measures will predict progression to diabetes better than the mere autoantibody-positivity.

1 型糖尿病 (T1D) 通过针对 65kD 亚型的自身抗体 (Ab) 进行预测 谷氨酸脱羧酶 (GAD65)、IA-2（一种蛋白酪氨酸磷酸酶样分子）和胰岛素。目的是揭示 GAD65、IA-2 和胰岛素自身抗体预测 T1D 的机制。总体假设是，随着个体向 TED 方向发展，抗体亲和力和表位特异性都会成熟，反映出抗原驱动的过程。目的是： 1. 检验以下假设：GAD65、IA-2 和胰岛素的亚型和同型自身抗体单独或组合可预测从胰岛自身免疫进展为 T1D。针对 GAD65、IA-2 和胰岛素的特定亚型和同种型自身抗体将作为进展或不进展为 T1D 的函数进行分析。 2. 检验以下假设：T1D 的进展与针对特定自身抗原表位的自身抗体成熟相关。使用 GAD65/67 杂合蛋白和竞争性表位特异性重组 Fab 进行亚型和亚型特异性测定，可将自身抗体成熟为特定表位 建立为进展或不进展为 T1D 的函数。 3. 检验 T1D 进展与表位特异性自身抗体的亲和力相关的假设。将通过针对 GAD65 和胰岛素独特表位的重组 Fab 的竞争测定来检查效价和自身抗体亲和力成熟度。自身抗体亲和力和亲和常数的测量将用于区分 T1D 的非进展者和进展者。项目 2 将与 a) 项目 1 合作，研究非进展者、进展者和新发 T1D 患者以及与自身抗体特征相关的 CD4 T 细胞分析； b) 项目 3 关于新发经典 T1D 患者与 1.5 型糖尿病受试者的比较以及 T 细胞增殖至多种胰岛细胞蛋白和细胞因子分泌模式，以及 c) 项目 4 对一般人群 T1D 风险受试者的 HLA 和非 HLA 遗传因素进行跟踪，直至胰岛自身免疫和 T1D。 Core B 将有助于对从产生 GAD65、IA2 和胰岛素单克隆抗体的杂交瘤细胞系中克隆的 Fab cDNA 进行测序。我们期望发现自身抗体成熟度测量比单纯自身抗体阳性更好地预测糖尿病的进展。