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: 8052888
• 负责人: Maki Nakayama
• 金额: $ 24.65万
• 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/8052888
• 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; 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）可能是主要靶点 NOD小鼠自发性糖尿病的发生和识别这种肽的T细胞 具有一个T细胞受体，专门共享保守的Valpha和Jalpha片段。我将定义使用 移植，具体的时间和位置的贡献胰岛素B：9-23在发展中的胰岛 自身免疫我还将直接测试基因组编码的T细胞受体链是 对NOD小鼠疾病和胰岛素B的T细胞库识别至关重要：9-23 肽。我建议创造NOD小鼠，具有假定的“不相关”的单个Jalpha 56片段， 具有与保守的Jalpha片段不同的序列（53/42），并对其进行开发评估 抗胰岛自身免疫建立了仅携带Jalpha 56的C57 BL/6小鼠，其中单个Jalpha是 用于创建免疫库，我们将使用速度来创建同源Jalpha 56 NOD小鼠， 同类技术我预测如果Jalpha 53/42保守序列是关键的， 对于仅具有“不相关”J α 56序列的NOD小鼠，不会产生自身免疫。如果预防 我们将从分子水平创造出具有单一Jalpha 56序列的小鼠 被改造成J α 53序列，我预测会恢复自身免疫在独立阶段，我 将定义赋予抗胰岛素自身免疫性的保守α链的序列， 分子逆转录技术用原始和修饰的α链T细胞产生逆转录基因小鼠 受体将使我们能够确定关键序列。我们将分子转移氨基酸盒， 将抗B：9-23致糖尿病克隆的保守的Valpha和Jalpha转化为“不相关的”对照α链， 评估我们是否可以创建抗胰岛素自身免疫，并将从对照转移氨基酸盒 α链转化为抗B：9-23致糖尿病α链以抑制或消除自身免疫。如果这 这条途径在NOD小鼠中至关重要，它将刺激寻找类似的人类关键途径。作为 作为一个长期目标，计划包括首先在小鼠模型中开发方法，并最终应用于 类似的逆转录和分子技术，从胰岛浸润的人T细胞受体， 伙计