STRUCTURE PEPTIDE/CONSERVED T CELL RECEPTOR DETERMINING DIABETES

决定糖尿病的结构肽/保守 T 细胞受体

• 批准号: 8009618
• 负责人: Maki Nakayama
• 金额: $ 24.9万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-04-01 至 2013-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8009618
• 关键词: Amino Acids; Antigens; Autoantibodies; Autoantigens; Autoimmunity; B-insulin; C57BL/6 Mouse; Cells; Conserved Sequence; Critical Pathways; Data; Development; Diabetes Mellitus; Disease; Future; Germ Lines; Goals; Human; Immune; Immunotherapy; Inbred NOD Mice; Infiltration; Insulin; Insulin-Dependent Diabetes Mellitus; Islets of Langerhans; Knock-in Mouse; Knock-out; Location; Methodology; Modeling; Molecular; Mus; Pancreas; Pathway interactions; Peptides; Phase; Prevention; Receptor Cell; Sequence Deletion; Specificity; Speed; Staging; System; T-Cell Receptor; T-Cell Receptors alpha-Chain; T-Lymphocyte; Techniques; Testing; Time; Transplantation; base; clinically relevant; congenic; diabetic patient; disorder prevention; endocrine pancreas development; genome sequencing; islet; laser capture microdissection; man; mouse development; mouse model; novel; peptide B; peptide structure; prevent; reconstitution; restoration

Our data indicates that insulin B chain amino acids 9 to 23 peptide (insulin B:9-23) may be a primary target essential for the initiation of spontaneous diabetes in NOD mice and that T cells recognizing this peptide have a T cell receptor that specifically shares conserved Valpha and Jalpha segments. I will define using transplants, the specific timing and location of the contribution of insulin B:9-23 in the development of islet autoimmunity. I will also directly test the hypothesis that genomically encoded T cell receptor chains are crucial for disease in the NOD mouse and for recognition by the T cell repertoire of the insulin B:9-23 peptide. I propose to create NOD mice having the presumed "irrelevant" single Jalpha 56 segment, which has different sequence from the conserved Jalpha segment (53/42), and to evaluate them for development of anti-islet autoimmunity. C57BL/6 mice bearing only Jalpha 56 were established where a single Jalpha is used to create the immune repertoire, and we will create congenic Jalpha 56 NOD mice using speed congenic techniques. I predict if the Jalpha 53/42 conserved sequences are critical, anti-insulin/islet autoimmunity will not develop for NOD mice having only the "irrelevant" Jalpha 56 sequence. If prevention occurs in mice with the single Jalpha 56, we will molecularly create mice with the single Jalpha 56 sequence modified to be a Jalpha 53 sequence, and I predict restoration of autoimmunity. For the independent phase, I will define sequences of the conserved alpha chain that confer anti-insulin autoimmunity with novel molecular retrogenic techniques. Producing retrogenic mice with the original and modified alpha chain T cell receptors will allow us to define crucial sequences. We will molecularly transfer amino acid cassettes from the conserved Valpha and Jalpha of anti-B:9-23 diabetogenic clones into a "irrelevant" control alpha chain to assess whether we can create anti-insulin autoimmunity, and will transfer amino acid cassettes from control alpha chain into anti-B:9-23 diabetogenic alpha chains to suppress or abrogate autoimmunity. If this pathway is critical in the NOD mouse, it will stimulate a search for an analogous human critical pathway. As a long-term goal, plans are included to first develop methodology in the mouse model and eventually apply similar retrogenic and molecular techniques to T cell receptors of man from pancreatic islet infiltration of man.

我们的数据表明，胰岛素B链氨基酸9至23肽（胰岛素B：9-23）可能是主要靶标 在NOD小鼠中开始自发糖尿病和识别该肽的T细胞至关重要 具有特异性共享保守的瓦尔法和墨西哥胡椒段的T细胞受体。我将使用 移植物，胰岛素B：9-23在胰岛发展中的贡献的特定时间和位置 自身免疫性。我还将直接检验以下假设：基因组编码的T细胞受体链是 胰岛素B：9-23 肽。我建议创建具有假定“无关”的单贾尔帕56段的点头小鼠， 具有与保守的贾尔法段不同的序列（53/42），并评估它们以进行开发 反刑事自身免疫。 C57BL/6小鼠仅设有Jalpha 56 用于创建免疫曲目，我们将使用速度创建Encenic Jalpha 56点头小鼠 先天技术。我预测墨西哥胡椒53/42保守的序列是否关键，抗胰岛素/小岛 对于只有“无关”的jalpha 56序列的NOD小鼠，自身免疫性不会发展。如果预防 发生在带有单一果酱56的小鼠中，我们将用单贾尔帕56序列产生小鼠 修改为jalpha 53序列，我预测自身免疫性的恢复。对于独立阶段，我 将定义保守的α链的序列，该序列允许新型抗胰岛素自身免疫性 分子后源技术。用原始和改良的α链T细胞产生肾后源性小鼠 受体将使我们能够定义关键序列。我们将从分子转移氨基酸盒 抗B的保守valpha和Jalpha：9-23个糖尿病的克隆进入“无关”的控制α链到 评估我们是否可以创建抗胰岛素自身免疫性，并将氨基酸盒从控制中转移 α链中的抗B链：9-23个糖尿病生成α链，以抑制或消除自身免疫性。如果这个 途径在点头小鼠中至关重要，它将刺激搜索类似的人类关键途径。作为 一个长期目标，包括在鼠标模型中首先开发方法并最终应用的计划 与胰岛的T细胞受体相似的后源性和分子技术类似 男人。