Modeling and Detecting Pathogens of Islet Autoimmunity

描述（由申请人提供）：DAISY研究和改进的 对针对胰岛素的自身免疫的理解以及 检测未知病原体（例如Skip Virgin：“哨兵免疫缺陷小鼠和 新获得的DNA和RNA的代表性差异分析）， 相信为寻找传染源提供了独特的机会 可能引发TIDM的发展。 我们建议在 平行的一般搜索传染性病原体和一个重点项目 集中在“致病性”胰岛素肽上。 我们的初步数据来自 DAISY研究表明，我们现在可以在出生时识别儿童（兄弟姐妹）， DR 3/4、DQ 8/2）的1型糖尿病患者，有激活风险 抗胰岛自身抗体表达在前两年内超过50% 生活 此外，我们最近发现， 遗传背景（例如基于Balb/c小鼠的MHC），我们可以很容易地 用特异性胰岛素肽诱导正常人抗胰岛素自身抗体 小鼠也被人类T细胞识别。 使用这种肽，结合 用病毒模拟物（poly-IC[polyinosinic-polycytodylic acid]），我们可以诱导 胰岛炎，并伴有胰岛免疫调节缺陷（转基因B7-1 表达），可诱导胰岛破坏和糖尿病。 发展 该模型中的糖尿病似乎严重依赖于遗传因素， 背景，诱发环境因素，以及“巧合”的时间 诱发因素 这为类型的“环境”诱导提供了一个模型 1糖尿病的发展，在目前的建议，我们将1。发展 方法和测试密集的前瞻性评价的儿童为 出现抗胰岛自身抗体。2.检测的发生 感染（干扰素α，2 ′，5 ′-寡腺苷酸合成酶， “哨兵免疫缺陷小鼠”，消减DNA和cDNA杂交）和 序列环境病原体“暂时”与外观的 抗胰岛自身抗体3.定义B：9-23肽的模拟表位， “随机”肽文库和B：9-23应答性T细胞克隆。4.测试 胰岛素，B：9-23个模拟表位，用于诱导胰岛素自身抗体， 胰岛炎和糖尿病。5.评价 具有B：9-23的感染性病原体的肽或模拟表位相关序列， 刺激人和小鼠的B：9-23反应性T细胞克隆的能力。6.测试 激活B：9-23反应性T细胞克隆的病原体相关肽， 在遗传易感鼠品系中的疾病诱导，并按照 特异性目的1、特异性病原体是否与B：9-23/模拟表位相关 糖尿病前期儿童的发病顺序和发病时间 产生了抗胰岛自身免疫

DESCRIPTION (provided by applicant): The DAISY study and improved understanding of autoimmunity directed against insulin and novel methods to detect unknown pathogens (e.g. Skip Virgin: "sentinel immunodeficient mice and representational difference analysis for newly acquired DNA and RNA) we believe provide unique opportunities to search for infectious agents potentially triggering the development of TIDM. We propose to pursue in parallel a general search for infectious pathogens and a focused project concentrating on a "pathogenic" insulin peptide. Our preliminary data from the DAISY study indicates we can now identify children at birth (siblings of patients with type 1 diabetes with DR3/4, DQ8/2) with a risk of activating anti-islet autoantibody expression exceeding 50% within the first two years of life. In addition, we have recently discovered that given an appropriate genetic background (e.g. based on the MHC of Balb/c mice) we can readily induce anti-insulin autoantibodies with a specific insulin peptide in normal mice that is also recognized by human T cells. Using this peptide, combined with a viral mimic (poly-IC[polyinosinic-polycytodylic acid]) we can induce insulitis, and with a immunoregulatory defect in the islets (transgenic B7-1 expression), can induce islet destruction and diabetes. Development of diabetes in this model appears to be critically dependent upon genetic background, inducing environmental factors, and the "coincidental" timing of inducing factors. This provides a model for "environmental" induction of type 1 diabetes development and in the current proposal we will 1. Develop methodology and test intensive prospective evaluation of children for the appearance of anti-islet autoantibodies. 2. Detect the occurrence of infections (assays for inteferon alpha, 2',5'- oligoadenylate synthetase, "sentinel immunodeficient mice", subtractive DNA and cDNA hybridization) and sequence environmental pathogens "temporally" associated with appearance of anti-islet autoantibodies. 3. Define mimotopes of the B:9-23 peptide using "random" peptide libraries and B:9-23 responsive T cell clones. 4. Test insulin, B:9-23 mimotopes, for the induction of insulin autoantibodies, insulitis, and diabetes in genetically susceptible murine strains. 5. Evaluate peptides of infectious pathogens with B:9-23 or mimotope related sequences for ability to stimulate B:9-23 reactive T cell clones of man and mouse. 6. Test pathogen related peptides that activate B:9-23 reactive T cell clones for disease induction in genetically susceptible murine strains and evaluate as in specific aim 1, whether specific pathogens with B:9-23/mimotope related sequences infect prediabetic children and the timing of appearance in children developing anti-islet autoimmunity.