Modeling and Detecting Pathogens of Islet Autoimmunity

描述(申请人提供)：DAISY研究和改进 对针对胰岛素的自身免疫的认识和治疗新方法 检测未知病原体(例如，Skip Virgin：“哨兵免疫缺陷小鼠和 对新获得的DNA和RNA的代表性差异分析 相信为寻找感染性病原体提供了独特的机会 潜在地触发了TIDM的发展。我们建议在 同时进行对传染性病原体的一般搜索和一个重点项目 专注于一种“致病”的胰岛素多肽。我们的初步数据来自 黛西的研究表明，我们现在可以在出生时识别孩子(兄弟姐妹 伴有DR3/4、DQ8/2)激活风险的1型糖尿病患者 抗胰岛自身抗体的表达在前两年超过50% 生活。此外，我们最近发现，鉴于适当的 遗传背景(例如，基于Balb/c小鼠的MHC)我们很容易 用特定的胰岛素多肽诱导正常人产生抗胰岛素自身抗体 小鼠也能被人类T细胞识别。使用此多肽，结合 用病毒模拟物(聚-IC[多肌苷-多细胞酸])我们可以诱导 胰岛炎伴免疫调节缺陷(转基因B7-1 表达)，可导致胰岛破坏和糖尿病。发展中的 这一模型中的糖尿病似乎严重依赖于基因 背景，诱发环境因素，以及“巧合”时机 诱发因素。这为“环境”类型的归纳提供了一种模型 1.糖尿病的发展，在目前的提案中我们将1.发展 对儿童进行前瞻性评估的方法学和测试强度 出现抗胰岛自身抗体。2.检测是否发生 感染(干扰素α，2‘，5’-寡腺苷合成酶， “哨兵免疫缺陷小鼠”、消减DNA和cDNA杂交)和 与出现的环境病原体“暂时”相关的序列 抗胰岛自身抗体。3.确定B：9-23肽的模拟表位 “随机”多肽库和B：9-23反应性T细胞克隆。4.测试 胰岛素，B：9-23模拟表位，用于诱导胰岛素自身抗体， 在遗传易感的小鼠品系中表现为岛炎和糖尿病。5.评估 具有B：9-23或模拟表位相关序列的感染性病原体的多肽 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多肽 遗传易感小鼠品系的致病作用及评价 特异靶1，特定病原体与B：9-23/模拟表位是否相关 感染糖尿病前期儿童的序列和儿童出现的时间 发展抗胰岛自身免疫。

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.