PROTEINASE MODULATION DURING T CELL-ENDOTHELIAL ADHESION

T 细胞内皮粘附过程中的蛋白酶调节

• 批准号: 2460025
• 负责人: JOSEPH A MADRI
• 金额: $ 32.25万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-08-01 至 2000-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2460025
• 关键词: SCID mouse; T lymphocyte; biological signal transduction; cell cell interaction; cell migration; cell type; endopeptidases; experimental allergic encephalomyelitis; extracellular matrix; human tissue; integrins; laboratory rabbit; leukocyte adhesion molecules; ligands; molecular cloning; multiple sclerosis; myelin basic proteins; northern blottings; tissue /cell culture; vascular endothelium; western blottings

During the inflammatory process T cell transmigration through the endothelial cell layer and migration into the underlying and surrounding extracellular matrix is initiated by T cell adhesion to the endothelium. T cell - endothelial cell adhesion is mediated by specific ligands resident on the surfaces of both the T cell and the endothelial cell, specifically, VLA-4 (a4beta1) on the T cell and VCAM-1 on the endothelial cell. We have demonstrated that engagement of this ligand pair evokes changes in 72 kDa gelatinase as well as plasminogen activator and plasminogen activator inhibitor-1 in both cell populations, consistent with the manifestation of an invasive phenotype in the adherent T cell population and an "activated" phenotype in the endothelial cells. Resultant proteolysis of basement membrane and interstitial matrix components is thought to facilitate T cell extravasation out of the affected vessel and toward the site of inflammation. In this proposal wee will identify and characterize selected adhesion-inducible T cell and endothelial cell proteinase/proteinase inhibitor systems and T cell surface adhesion molecule expression. We will investigate and identify the signal transduction systems triggered by engagement of the VLA-4/VCAM-1 ligand pair in these two cell types. This will be accomplished utilizing an in vitro culture model utilizing cloned murine T cell clones specific for myelin basic protein and selected human T cell clones isolated from multiple sclerosis patients; an in vivo adoptive transfer murine model for experimental allergic encephalomyelitis (EAE) and a SCID mouse containing grafted human skin and reconstituted with human peripheral lymphocytes. A variety of methodologies will be employed including cell culture, zymography, reverse zymography, gene products, histology, immunohistochemistry and an animal model of EAE. These experiments will lead to a better understanding of T cell migration through and interaction with local extracellular matrix and the development of new and novel therapies directed at modulating selected proteinase/proteinase inhibitor cascade systems in the inflammatory processes of arthritis, vasculitis, organ rejection.

在炎症过程中，T细胞通过 内皮细胞层和向下层和周围的迁移 细胞外基质是由T细胞与内皮细胞的黏附启动的。 T细胞与血管内皮细胞的黏附是由特异性配体介导的 在T细胞和内皮细胞的表面，具体地说， T细胞表达VLA-4(A4beta1)，内皮细胞表达VCAM-1。我们有 证明了该配体对的结合引起72 kDa的变化 明胶酶、纤溶酶原激活剂和纤溶酶原激活剂 抑制物-1在两个细胞群中的表达，与 贴壁T细胞群体中的一种侵袭性表型和一种“激活” 内皮细胞的表型。所产生的基底区蛋白分解 膜和间质基质成分被认为促进T细胞 从受影响的血管渗出并向血管部位渗出 发炎。在本建议书中，我们将识别和描述选定 黏附诱导T细胞与内皮细胞蛋白酶/蛋白水解酶 抑制系统与T细胞表面黏附分子的表达。我们会 调查并确定由以下因素触发的信号转导系统 VLA-4/VCAM-1配基对在这两种细胞类型中的接合。这 将通过使用克隆的体外培养模型来完成 小鼠髓鞘碱性蛋白和精选人T细胞克隆 从多发性硬化症患者体内分离的T细胞克隆 实验性变态反应性脑脊髓炎过继转移小鼠模型的建立 (EAE)和含有移植的人皮肤的SCID小鼠 人外周血淋巴细胞。将采用多种方法 包括细胞培养，酶谱图，反向酶谱图，基因产品， 组织学、免疫组织化学和EAE动物模型。这些 实验将有助于更好地理解T细胞通过 与局部细胞外基质的相互作用和新的 以及针对调节选定的蛋白酶/蛋白水解酶的新疗法 关节炎炎症过程中的抑制物级联系统， 脉管炎，器官排斥。