DYNAMICS OF PERIPHERAL T CELL REPERTOIRE

外周 T 细胞库的动力学

• 批准号: 6472100
• 负责人: STANISLAV VUKMANOVIC
• 金额: $ 18.84万
• 依托单位: NEW YORK UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-01 至 2007-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6472100
• 关键词: MHC class I antigen; T cell receptor; antigen presentation; autoantigens; autoimmunity; cytotoxic T lymphocyte; dendritic cells; fibroblasts; genetically modified animals; laboratory mouse; leukopoiesis; liver cells; mass spectrometry; thymus

DESCRIPTION (provided by the applicant): The focus of the application is centered on recently discovered process of MHC class I-mediated peripheral selection of CD84 T cells. Peripheral selection is of paramount importance for: 1) maintaining the peripheral CD8+ T cell pool; 2) modifying the available peripheral CD8+ T cell repertoire generated in the thymus; and 3) possibly determining the length of a life of CD8+ T cells in the periphery. Uncovering the laws governing the process of peripheral selection will open enormous possibilities of manipulating the peripheral T cell repertoire. (Specific Aim 1) To determine whether the potential to promote peripheral selection is cell-type specific, selective expression of MHC class 1 on fibroblasts or dendritic cells will be achieved by making transgenic mice expressing the TAP 1 gene under the fibroblast-1 (FSP-1) or dendritic cell- (CD11c) specific promoters, crossing the transgenic mice to the TAP1-deficient background, and testing the ability of fibroblasts and dendritic cells to promote peripheral selection. (Specific Aim 2) To determine whether the ligands involved in the peripheral selection and positive thymic selection are identical MHC (miss)matching fetal liver and thymic epithelium will be grafted to observe the effects of matching MHC molecules in the thymus and periphery on repopulation of periphery by CD8+ T cells. In the second approach, it will be tested whether candidate self MHC-associated peptides that induce positive thymic selection of particular TCR transgenic thymocytes can also induce peripheral selection of the same TCR transgenic CD8+ cells. (Specific Aim 3) To identify true self peptides that induce peripheral selection, gene knock-out mouse strains will be immunized to produce CD8+ T cells specific for peptides derived from the missing gene. These CD8+ T cells will be used as a tool for detection of peptide presentation in wild-type mice. An alternative strategy will consist of mass spectrometry analysis of peptides eluted from immunoprecipitated MHC class I molecules. Detailed knowledge gained from the proposed experiments will be valuable for vaccine development and immunomodulation therapy in organ transplantation, autoimmune diseases and cancer.

描述（申请人提供）：申请的重点是 集中于最近发现的MHC I类介导的外围的过程 选择CD84 T细胞。外围选择至关重要： 1）维持外围CD8+ T细胞池； 2）修改可用的 胸腺中产生的外围CD8+ T细胞库； 3）可能 确定周围CD8+ T细胞寿命的长度。发现 管理外围选择过程的法律将开放巨大 操纵周围T细胞库的可能性。 （特定目标 1）确定促进外围选择的潜力是否是 成纤维细胞上MHC 1类的细胞类型特异性表达或 树突状细胞将通过使表达TAP 1的转基因小鼠来实现 基因在成纤维细胞1（FSP-1）或树突状细胞（CD11C）特异性下 发起人，将转基因小鼠跨越TAP1缺陷背景，然后 测试成纤维细胞和树突状细胞促进周围的能力 选择。 （特定目的2）确定是否参与 外围选择和阳性胸腺选择是相同的MHC （错过）匹配胎儿肝脏和胸腺上皮将被嫁接以观察 在胸腺和外围匹配的MHC分子对重生的影响 CD8+ T细胞的周围。在第二种方法中，将测试是否 候选自我MHC相关的肽，诱导阳性胸腺选择 特定的TCR转基因胸腺细胞也可以诱导外围选择 相同的TCR转基因CD8+细胞。 （特定目的3）确定真实的自我 诱导周围选择的肽，基因敲除小鼠菌株将是 免疫以产生针对源自肽的肽的CD8+ T细胞 缺失基因。这些CD8+ T细胞将用作检测的工具 野生型小鼠的肽表现。另一种策略将包括 从免疫沉淀的MHC类洗脱的肽的质谱分析 我的分子。从提出的实验中获得的详细知识将是 对于器官中的疫苗开发和免疫调节疗法来说很有价值 移植，自身免疫性疾病和癌症。