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: 2227386
• 负责人: JOSEPH A MADRI
• 金额: $ 30.83万
• 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/2227386
• 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 内皮细胞层并迁移到底层和周围 细胞外基质是由 T 细胞粘附到内皮细胞启动的。 T细胞-内皮细胞粘附是由驻留的特定配体介导的 在 T 细胞和内皮细胞的表面上，具体来说， T 细胞上的 VLA-4 (a4beta1) 和内皮细胞上的 VCAM-1。 我们有 证明该配体对的接合引起 72 kDa 的变化 明胶酶以及纤溶酶原激活剂和纤溶酶原激活剂 两个细胞群中都有抑制物-1，与表现一致 贴壁 T 细胞群中的侵袭性表型和“激活” 内皮细胞的表型。 基底蛋白水解作用 膜和间质基质成分被认为有利于 T 细胞 从受影响的血管外渗到受影响的部位 炎。 在本提案中，我们将确定并描述所选的 粘附诱导 T 细胞和内皮细胞蛋白酶/蛋白酶 抑制剂系统和T细胞表面粘附分子表达。 我们将 研究并识别由以下因素触发的信号转导系统 VLA-4/VCAM-1 配体对在这两种细胞类型中的结合。 这 将利用体外培养模型利用克隆来完成 对髓磷脂碱性蛋白和选定的人类具有特异性的鼠 T 细胞克隆 从多发性硬化症患者体内分离出的 T 细胞克隆；体内的 实验性过敏性脑脊髓炎过继转移小鼠模型 (EAE) 和含有移植人类皮肤并用重组的 SCID 小鼠 人外周淋巴细胞。 将采用多种方法 包括细胞培养、酶谱、反向酶谱、基因产物、 EAE 的组织学、免疫组织化学和动物模型。 这些 实验将有助于更好地了解 T 细胞迁移 和与局部细胞外基质的相互作用以及新的开发 以及针对调节选定蛋白酶/蛋白酶的新疗法 关节炎炎症过程中的抑制剂级联系统， 血管炎、器官排斥反应。