MAC-1 AND P150,95 AVIDITY CHANGES IN TRANSFECTED CELLS

中性粒细胞和单核细胞在炎症组织上的迁移和黏附 对正常的免疫功能是必不可少的，是早期的重要组成部分 肾小球肾炎和移植肾急性排斥反应。白血球 整合素Mac-1和p150，95是骨髓单核细胞 相互黏附，与内皮细胞和人肾小球系膜细胞黏附。 这种结合功能是由细胞刺激调节的， 化学引诱剂或佛波酯；涉及的第二个信使 仍有待定义。本项目的目标是研究 或细胞刺激后Mac-1和p150、95的亲和力 表征这两个分子的配基的组织分布。 我们将使用的策略是：1)转染人mac-1和p150，95 转化为小鼠杂交瘤T细胞系(其中一些杂交瘤T细胞系中的一些细胞缺乏第二 Messenger Products)，2)分析Mac-1和p150.95亲和力的变化 经细胞刺激后的转基因细胞，以及3)评估与 转染人中性粒细胞/内皮细胞、细胞基质 成分和冰冻组织切片。小鼠T细胞杂交瘤的应用 由于转染体将允许分析整合素亲和力的变化 用抗原刺激细胞，以及使用基因缺失的杂交瘤 第二信使的生产将允许我们剖析所涉及的路径 在整合素结合的变化中。此外，其他人的互动 物种特有的表面分子将通过使用 小鼠T细胞杂交瘤。

Neutrophil and monocyte migration and adherence to inflamed tissue are essential for normal immune function and are important components of early glomerulonephritis and acute renal allograft rejection. The leukocyte integrins, Mac-1 and p150,95, are essential for myelomonocytic cells to adhere to each other, to endothelial cells and to human mesangial cells. This binding function is regulated by cellular stimulation with chemoattractants or by phorbol esters; the second messengers involved remain to be defined. The goals of this project are to study the binding or avidity of Mac-1 and p150,95 after cellular stimulation and to characterize the tissue distribution of the ligands for both molecules. The strategy we will use will be to: 1) transfect human Mac-1 and p150,95 into mouse hybridoma T cell lines (some of which are deficient in second messenger production), 2) analyze changes in Mac-1 and p150.95 avidity in transfected cells after cellular stimulation and, 3) assess the binding of transfectants to human neutrophils/endothelial cells, cell matrix components and frozen tissue sections. The use of murine T cell hybridomas as transfectants will permit an analysis of integrin avidity change after cellular stimulation with antigen, and the use of hybridomas deficient in second messenger production will allow us to dissect the pathways involved in the change in integrin binding. In addition, interactions by other surface molecules that are species specific will be avoided by the use of mouse T-cell hybridomas.