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9-23肽(胰岛素B：9-23)可能是主要的靶标 NOD小鼠自发性糖尿病的发生和识别该肽的T细胞 有一个T细胞受体，专门分享保守的Valpha和Jpha片段。我将使用以下内容定义 移植，胰岛素B：9-23在胰岛发育中作用的具体时间和位置 自身免疫力。我还将直接测试这样一个假设，即基因编码的T细胞受体链是 对NOD小鼠的疾病和T细胞识别胰岛素B：9-23至关重要 多肽。我建议创造NOD小鼠，该小鼠具有推定的“无关”的单个Jalpha56片段，该片段 具有与保守的Jpha片段(53/42)不同的序列，并对它们进行发育评估 抗胰岛自身免疫力。只携带Jalpha56的C57BL/6小鼠被建立在只有一个Jpha56的C57BL/6小鼠 用来创建免疫系统，我们将使用SPEED创建同源的Jalpha56nod小鼠 同源基因技术。我预测Jalpha53/42保守序列是否是关键的抗胰岛素/胰岛 仅有Jalpha56“无关”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Jalpha56序列的小鼠中，我们将从分子上创建具有单个Jalpha56序列的小鼠 被修改为Jalpha53序列，我预测自身免疫功能恢复。对于独立阶段，我 将定义保守的阿尔法链序列，使抗胰岛素自身免疫具有新的 分子回溯技术。用原始和修饰的阿尔法链T细胞制备逆转录基因小鼠 受体将允许我们定义关键序列。我们将从分子上转移氨基酸盒 抗-B：9-23致糖尿病克隆的保守的Valpha和Jpha进入一个“无关”的控制阿尔法链 评估我们是否可以创造抗胰岛素自身免疫，并将从控制组转移氨基酸盒 α链进入抗B：9-23致糖尿病的阿尔法链，以抑制或消除自身免疫。如果这个 途径在NOD小鼠中是关键的，它将刺激人们寻找类似人类的关键途径。AS 作为一个长期目标，计划首先在小鼠模型中开发方法学并最终应用 类似逆转录和分子生物学技术的人T细胞受体从胰岛浸润性 天哪。