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）可能是主要目标 对于 NOD 小鼠自发性糖尿病的发生至关重要，并且 T 细胞识别该肽 具有特异性共享保守 Valpha 和 Jalpha 片段的 T 细胞受体。我将定义使用 移植，胰岛素 B:9-23 在胰岛发育中贡献的具体时间和位置 自身免疫。我还将直接检验基因组编码的 T 细胞受体链的假设 对于 NOD 小鼠的疾病以及胰岛素 B:9-23 T 细胞库的识别至关重要 肽。我建议创建具有假定“不相关”的单个 Jalpha 56 片段的 NOD 小鼠，这 与保守的 Jalpha 片段 (53/42) 具有不同的序列，并评估它们的发育 抗胰岛自身免疫。建立仅携带 Jalpha 56 的 C57BL/6 小鼠，其中单个 Jalpha 是 用于创建免疫组库，我们将使用速度创建同类 Jalpha 56 NOD 小鼠 同类技术。我预测 Jalpha 53/42 保守序列是否至关重要，抗胰岛素/胰岛 仅具有“不相关”Jalpha 56 序列的 NOD 小鼠不会产生自身免疫。如果预防 发生在具有单个 Jalpha 56 的小鼠中，我们将通过分子方法创建具有单个 Jalpha 56 序列的小鼠 修改为 Jalpha 53 序列，我预测自身免疫会恢复。对于独立阶段，我 将定义保守α链的序列，赋予抗胰岛素自身免疫性新的 分子逆转录技术。使用原始和改良的 α 链 T 细胞生产逆转录小鼠 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 将抗 B:9-23 糖尿病克隆的保守 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，并将从对照中转移氨基酸盒 α 链转化为抗 B:9-23 糖尿病性 α 链以抑制或消除自身免疫。如果这个 该通路在 NOD 小鼠中至关重要，它将刺激人们寻找类似的人类关键通路。作为 作为一个长期目标，计划首先在小鼠模型中开发方法并最终应用 与人类胰岛浸润的 T 细胞受体相似的逆转录和分子技术 男人。