STRUCTURAL BASIS OF CD59 AND CD58 SIGNALING TO T CELLS

CD59 和 CD58 向 T 细胞发出信号的结构基础

• 批准号: 2638013
• 负责人: ALFRED LM BOTHWELL
• 金额: $ 30.36万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-01-01 至 2001-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2638013
• 关键词: 3T3 cells; CD antigens; CHO cells; SCID mouse; T lymphocyte; anergy; antigen antibody reaction; antigen presenting cell; cell adhesion molecules; cell cell interaction; histocompatibility; human subject; leukocyte adhesion molecules; phlebotomy; protein structure function; tissue /cell culture; transfection; transplantation immunology; vascular endothelium; xenotransplantation

DESCRIPTION (Adapted from the Applicant's Abstract): This is a competitive renewal application. The goals of this project are to characterize specific costimulatory pathways invoked during allo and xeno immune responses to endothelium. Studies have defined common properties of the responses to human and porcine ECs but also have identified a major important difference regarding the expression of B7. There are substantial sequence differences in the cell surface antigens between these antigens and their contribution to signaling via ligand interactions may differ significantly. The response of human T cells is much stronger to porcine ECs than human allogeneic ECs. This is a consequence of direct recognition of MHC class I and class II antigens as well as the presence of pB7.2 on porcine ECs. The role of CD58 and CD59 in the CD2 pathway and pB7.2 in the CD28 pathway in xeno responses will be studied. The activation of human T cells will be assessed using complementary approaches. The contribution of distinct ligand interactions on transfected CHO cells individually or in combination will be examined. This will include normal and mutant forms of human CD58 and CD59 antigens as well as normal forms of these homologous porcine antigens. In addition, more complex antigen presenting cells will be studied after removing defined antigens. Several strategies will be employed to specifically inhibit the cell surface expression of SLA antigens and costimulatory molecules (pB7.2, CD59, CD58) on human and porcine ECs. Both primary as well as recall responses will be evaluated. The ability of human T cells to recognize SLA antigens will be studied by expressing these antigens on transfected cells. Recognition of SLA antigens in the absence of a costimulatory signal may result in induction of anergic human T cells. Transfections of NIH3T3 cells with SLA antigens plus additional costimulatory molecules will be utilized to define the requirements for breaking anergy. Comparisons between human and porcine systems will be made in order to define the critical pathways that could be targets for facilitating tissue transplants. Finally, the applicants will utilize an in vivo model in which the immune system of SCID mice has been reconstituted with human cells. The immunogenic properties of the transfected porcine ECs that have demonstrated altered recognition properties in culture will be evaluated in this model after formation of synthetic vascular networks in collagen gels.

描述（改编自申请人的摘要）：这是一个有竞争力的 续订申请。 该项目的目标是描述具体的 同种异体和异种免疫反应期间调用的共刺激途径 内皮细胞。 研究已经定义了响应的共同属性 人类和猪的 ECs 但也发现了一个重大的重要差异 关于B7的表达。 存在显着的序列差异 细胞表面抗原之间的关系以及这些抗原的贡献 通过配体相互作用的信号传导可能存在显着差异。 人类 T 细胞对猪 EC 的反应比人类 T 细胞强得多 同种异体EC。 这是直接识别 MHC I 类的结果 和 II 类抗原以及猪 EC 上 pB7.2 的存在。 这 CD58和CD59在CD2途径中的作用以及pB7.2在CD28途径中的作用 将研究异种反应。 人类T细胞的激活将是 使用补充方法进行评估。 不同配体相互作用对转染 CHO 细胞的贡献 将单独或组合地进行检查。 这将包括正常 和人类 CD58 和 CD59 抗原的突变形式以及正常形式 这些同源猪抗原。 此外，更复杂的抗原 去除确定的抗原后将研究呈递细胞。 一些 将采用策略来特异性抑制细胞表面 SLA 抗原和共刺激分子（pB7.2、CD59、CD58）的表达 对人和猪的 EC。 主要反应和回忆反应都将是 评价。 将研究人类 T 细胞识别 SLA 抗原的能力 在转染细胞上表达这些抗原。 SLA 抗原的识别 在缺乏共刺激信号的情况下可能会导致无能的诱导 人类 T 细胞。 用 SLA 抗原转染 NIH3T3 细胞 额外的共刺激分子将被用来定义 打破无能的要求。 人类与猪的比较 将建立系统来定义可能的关键路径 促进组织移植的目标。 最后，申请人将 利用体内模型，其中 SCID 小鼠的免疫系统已被 用人体细胞重建。 的免疫原性特性 转染的猪 EC 已表现出识别改变 在形成后，将在该模型中评估培养物的特性 胶原凝胶中的合成血管网络。