PROTEINASE MODULATION DURING T CELL-ENDOTHELIAL ADHESION

在炎症过程中，T细胞通过 内皮细胞层和迁移到下面和周围 细胞外基质是通过T细胞粘附到内皮而启动的。 T细胞-内皮细胞粘附是由细胞内的特异性配体介导的， 在T细胞和内皮细胞的表面，特别是， T细胞上的VLA-4（α 4 β 1）和内皮细胞上的VCAM-1。 我们有 证明了这种配体对的参与引起72 kDa的变化， 明胶酶以及纤溶酶原激活物和纤溶酶原激活物 抑制剂-1在两个细胞群中，与 粘附性T细胞群中的侵袭性表型和“活化”T细胞群中的侵袭性表型。 内皮细胞中的表型。 基底蛋白水解 膜和间质基质成分被认为有助于T细胞 外渗出受影响的血管，并向部位 炎症 在本提案中，我们将确定和描述选定的 粘附诱导T细胞和内皮细胞蛋白酶 抑制剂系统和T细胞表面粘附分子表达。 我们将 研究并确定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接合。 这 将利用体外培养模型完成，所述体外培养模型利用克隆的 特异于髓鞘碱性蛋白的鼠T细胞克隆和选择的人T细胞克隆 从多发性硬化症患者中分离的T细胞克隆;体内 实验性变态反应性脑脊髓炎小鼠过继转移模型 (EAE)以及含有移植的人皮肤并用 人外周血淋巴细胞 将采用各种方法 包括细胞培养、酶谱、反向酶谱、基因产物 组织学、免疫组织化学和EAE动物模型。 这些 实验将导致更好地理解T细胞迁移， 与局部细胞外基质的相互作用以及新的 以及针对调节选择的蛋白酶/蛋白酶的新疗法 抑制剂级联系统在关节炎的炎症过程中， 血管炎器官排斥

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.