AUTOANTIBODY ANALYSIS FOR A BETTER PREDICTION OF IDDM

自身抗体分析可更好地预测 IDDM

• 批准号: 6916759
• 负责人: AKE LERNMARK
• 金额: $ 13.43万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-04-01 至 2008-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6916759
• 关键词: antigen antibody reaction; antigen presentation; autoantibody; bioassay; clinical research; enzyme linked immunosorbent assay; flow cytometry; gene expression; glutamate decarboxylase; histocompatibility typing; human subject; immunoglobulin structure; insulin; insulin dependent diabetes mellitus; pathologic process; polymerase chain reaction; prognosis; protein binding; protein isoforms; protein tyrosine phosphatase; recombinant proteins; western blottings

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）由针对65KD同工型的自身抗体（AB）预测 谷氨酸脱羧酶（GAD65），IA-2，-A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，随着个体的发展，抗体的亲戚和表位特异性都成熟，以反映抗原驱动过程的TED。目的是：1。检验以下假设：单独或组合中的亚型和同型自身抗体对GAD65，IA-2和胰岛素单独或组合预测从胰岛自身免疫到T1D的进展。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非促进T1D的函数分析。 2。为了测试以下假设：对T1D的进展与针对特定自身抗原表位的自身抗体成熟有关。使用GAD65/67混合蛋白和竞争表位特异性重组FAB，使用同工和亚型特异性测定 建立是进展或非促进t1d的函数。 3。检验以下假设，即对T1D的进展与表位特异性自身抗体的亲和力有关。滴度和自身抗体亲和力成熟将在与重组Fab的竞争测定中检查，以对GAD65和胰岛素的独特表位进行检查。自身抗体亲和力常数的度量将用于区分非传播者，从进度者到T1D。项目2将与A）与自身抗体特征有关的非培训器，进度者和新发作T1D患者以及CD4 T细胞分析的项目1合作； b）与1.5型糖尿病受试者和T细胞增殖与多种胰岛细胞蛋白和细胞因子分泌模式相比，针对新发作的经典T1D患者的项目3和c）Project 4 HLA和T1D风险的一般人群中的项目4 HLA和非HLA遗传因素，直到胰岛自身免疫性和T1D。核B将对从产生GAD65，IA2和胰岛素产生单克隆抗体的杂交瘤细胞系克隆的序列Fab cDNA发挥作用。我们希望发现自身抗体成熟度措施将比单纯的自身抗体积极性更好地预测糖尿病的进展。