SYNTHESIS AND EXPRESSION OF THE THROMBOMODULIN ANTIGEN

血栓调节蛋白似乎是一种内皮细胞特异性膜结合蛋白， 细胞表面蛋白 最近的研究表明， 在原代培养物中表达血栓调节蛋白活性， 细胞系，甚至在多次传代后（12）。 因此， 血栓调节蛋白是由内皮细胞合成的 研究 将开始证明内皮细胞合成 血栓调节蛋白 血栓调节蛋白活性的合成和表达将 用对数期和汇合后的细胞进行研究。 的 血栓调节蛋白抗原的表达也将被研究， 与细胞周期和体外细胞老化的关系。 血栓调节蛋白活性在连续培养中的保留将是 研究了与其他内皮细胞标志物丢失的关系， 作为因子VIII抗原和血管紧张素转换酶。 的 将通过以下方法研究血栓调节蛋白的超微结构定位： 铁蛋白结合抗血栓调节蛋白免疫电子细胞化学 抗体或通过间接酶免疫细胞化学技术。 的 将研究血栓调节蛋白在整个脉管系统中的分布 通过免疫荧光和比较将在大与 微血管以及不同组织部位的微血管之间 (i.e.,脑对肺对心脏对等等）。 这些研究将提供 血栓调节蛋白器官特异性的定性信息 分布 其他类型的细胞也可能具有血栓调节蛋白 相关抗原 特别令人感兴趣的是， 血液和骨髓，将通过流式细胞术使用 抗血栓调节蛋白抗体（多克隆和单克隆）。 这些研究将 提供必要的资料，以启动关于机制的研究 控制血栓调节蛋白的生物合成、抗原的表达和 调节血栓调节蛋白的生物活性。

Thrombomodulin appears to be an endothelial cell specific membrane bound cell surface protein. Recent studies have shown that rabbit endothelium expresses thrombomodulin activity in primary cultures and within the same cell lines even after multiple passages (12). Therefore it appears likely that thrombombomodulin is synthesized by the endothelial cell. Studies will be initiated to demonstrate endothelial cell synthesis of thrombomodulin. Synthesis and expression of thrombomodulin activity will be studied with cells in log phase and following confluence. The expression of thrombomodulin antigen will also be studied in relationship to the cell cycle and in vitro cell aging using flow cytometry procedures. The retention of thrombomodulin activity in continuous culture will be studied in relationship to the loss of other endothelial cell markers such as Factor VIII antigen and angiotensin converting enzyme. The ultrastructural localization of thrombomodulin will be studied by immuno-electron-cytochemistry using ferritin conjugated antithrombomodulin antibodies or by indirect enzyme immunocytochemical techniques. The distribution of thrombomodulin throughout the vasculature will be studied by immunofluorescence and comparisons will be made between large vs. microvessels as well as between microvessels at different tissue sites (i.e., brain vs. lung vs. heart vs. etc.). These studies will provide qualitative information about the organ specificity of thrombomodulin distribution. It is possible that other cell types may have thrombomodulin associated antigen. Of particular interest are the formed elements of the blood and bone marrow which will be investigated by flow cytometry using anti-thrombomodulin antibodies (poly-and monoclonal). These studies will provide the information necessary to initiated studies on the mechanisms controlling thrombomodulin biosynthesis, expression of the antigen and regulation of thrombomodulin's biological activity.