FUNCTIONS OF CD40L IN ENDOTHELIAL, FIBROBLAST, AND T CELL REGULATION

CD40L 在内皮细胞、成纤维细胞和 T 细胞调节中的功能

• 批准号: 6201304
• 负责人: HONG JIANG
• 金额: $ 14.46万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-09-01 至 2001-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6201304
• 关键词: CD40 molecule; T lymphocyte; antigen presentation; antigen presenting cell; autoimmune disorder; cell adhesion molecules; cell cell interaction; cell cycle; clinical research; density gradient ultracentrifugation; enzyme linked immunosorbent assay; fibroblasts; flow cytometry; hemostasis; human subject; immunocytochemistry; immunopathology; immunoregulation; laboratory mouse; leukocyte activation /transformation; mixed tissue /cell culture; receptor expression; vascular endothelium

We propose to test the hypothesis that interactions mediated by T cell receptors (TCRs) expressed on different T cell subsets play pivotal roles in immunoregulation and are directly involved in the control of autoimmune disease. In previous studies we showed that CD8+ T cells in vivo control the outgrowth of CD4+ cells mediating experimental allergic encephalomyelitis (EAE). Moreover, we showed that CD8+T cells also participate in the in vivo downregulation of CD4+Vbeta8+T cells following SEB administration. We investigated these CD4/CD8 interactions in vitro and observed that following SEB administration CD8+Tcells emerge which preferentially kill activated CD4+Vbeta8+ but not CD4+Vbeta8- T cells in vitro. This TCR Vbeta specific cytotoxicity was dependent on beta 2 microglobulin and was inhibited by antisera specific for Qa-1 but not by antibody to MEC class 1-a. This data suggest that the specificity of immune regulation involves recognition of TCR Vbeta determinants and Qa-1 molecules expressed on activated CD4+T cells. Recently, we extended these findings to show that CD8+ T cells isolated from CD4+Vbeta8+ T cell vaccinated mice specifically lyse a human B cell line cotransfected with both murine TCR Vbeta8 and Qa-1 cDNA, but not Bcells transfected with either TCR or Qa-1 cDNA alone. Furthermore, we showed that vaccinating naive mice with activated CD4+Vbeta8+ T cells specifically enhance SEB induced in vivo deletion of CD4+Vbeta8+ T cells. These in vivo and in vitro data support the idea that at least one consequence of T cell vaccination is the induction of CD8+ Vbeta specific CTL which downregulate immune responses. Because T cell vaccination has been used to ameliorate autoimmune disease in a variety of animal models and in humans, the overall goals of this project are to further define the cellular and molecular mechanisms involved in T cell vaccination. We will first confirm the observation that TCR Vbeta specific CD8+ regulatory T cells are induced by T cell vaccination. We will then extend our analysis to the EAE model to determine if Vbeta specific CD8+ T cells also mediate the protective effect of T cell vaccination against EAE. We will also determine if the molecular structure recognized by these regulatory CD8+ T cells is the TCR Vbeta chain or peptide(s) derived from the TCR Vbeta chain associated with Qa-1 molecules. Finally, we will directly determine if adoptive transfer of T cell vaccination induced TCR Vbeta specific CD8+ T cells regulate EAE in vivo.

我们建议检验T细胞介导的相互作用的假设 在不同T细胞子集上表达的受体（TCR）扮演着关键作用 在免疫调节中，直接参与自身免疫的控制 疾病。 在先前的研究中，我们表明CD8+ T细胞体内对照 CD4+细胞的产物介导了实验过敏 脑脊髓炎（EAE）。 此外，我们表明CD8+T细胞也 参与CD4+VBETA8+T细胞的体内下调 SEB管理。 我们在体外研究了这些CD4/CD8相互作用 并观察到SEB给药后CD8+TCELLS出现了 优先杀死活化的CD4+VBETA8+，而不是CD4+VBETA8- T细胞 体外。 这种TCR VBETA特异性细胞毒性取决于β2 微球蛋白，并被特定于QA-1的抗血清抑制，但不是 MEC 1-A类的抗体。 这些数据表明 免疫调节涉及识别TCR VBETA决定因素和QA-1 在活化的CD4+T细胞上表达的分子。 最近，我们扩展了这些 发现从CD4+ VBETA8+ T细胞中分离出的CD8+ T细胞 疫苗接种的小鼠特异性裂解了与人类B细胞系共转染 鼠TCR VBETA8和QA-1 cDNA均未转染 单独使用TCR或QA-1 cDNA。 此外，我们证明了接种疫苗 活化的CD4+ VBETA8+ T细胞的天真小鼠特异性增强了SEB 诱导的CD4+ VBETA8+ T细胞的体内缺失。 这些体内和 体外数据支持至少T细胞后果的想法 疫苗接种是CD8+ VBETA特异性CTL的诱导 免疫反应。 因为T细胞疫苗接种已用于改善 各种动物模型和人类中的自身免疫性疾病，总体 该项目的目标是进一步定义细胞和分子 涉及T细胞疫苗的机制。 我们将首先确认 观察到TCR VBETA特异性CD8+调节T细胞是由 T细胞疫苗接种。 然后，我们将分析扩展到EAE模型 确定VBETA特异性CD8+ T细胞是否也介导了保护效果 针对EAE的T细胞疫苗接种。 我们还将确定分子是否 这些调节性CD8+ T细胞识别的结构是TCR VBETA 从与QA-1相关的TCR VBETA链中得出的链或肽 分子。 最后，我们将直接确定t的收养转移是否 细胞疫苗接种诱导TCR VBETA特异性CD8+ T细胞调节EAE 体内。