Yang C, Jones J L, Barnum S R
Department of Microbiology, University of Alabama at Birmingham.
J Neuroimmunol. 1993 Sep;47(2):123-32. doi: 10.1016/0165-5728(93)90022-q.
In this report, we have shown the expression of the complement regulatory proteins decay-accelerating factor (DAF, CD55), membrane cofactor protein (MCP, CD46) and CD59 on human D54-MG astroglioma cells by several methods, including immunofluorescence, flow cytometry and Western blotting and Northern blot analysis. These studies demonstrate that all three proteins are structurally and antigenically similar to their counterparts expressed on HepG2 and SW480 cells (hepatocyte and epithelial cell lines, respectively). D54-MG cells express mRNA for all three proteins of the appropriate size(s). The phosphatidylinositol-specific enzyme, PIPLC, cleaved DAF from the surface of D54-MG cells, demonstrating that DAF is linked by a glycophospholipid anchor as has been shown for other cell types. Flow cytometry demonstrates that primary rat astrocytes also constitutively express all three regulatory proteins. These data are the first to demonstrate the expression of CD59 on astrocytes, and the presence of all three regulatory proteins on astrocytes suggests that regulation of complement activation in the central nervous system is important in neural host defense mechanisms.
在本报告中,我们通过多种方法,包括免疫荧光、流式细胞术、蛋白质印迹法和Northern印迹分析,展示了补体调节蛋白衰变加速因子(DAF,CD55)、膜辅因子蛋白(MCP,CD46)和CD59在人D54-MG星形胶质瘤细胞上的表达。这些研究表明,这三种蛋白在结构和抗原性上与其在HepG2和SW480细胞(分别为肝细胞系和上皮细胞系)上表达的对应蛋白相似。D54-MG细胞表达所有三种大小合适的蛋白的mRNA。磷脂酰肌醇特异性酶PIPLC从D54-MG细胞表面裂解DAF,表明DAF如在其他细胞类型中所示,是通过糖磷脂锚连接的。流式细胞术表明原代大鼠星形胶质细胞也组成性地表达所有三种调节蛋白。这些数据首次证明了CD59在星形胶质细胞上的表达,并且星形胶质细胞上所有三种调节蛋白的存在表明补体激活的调节在中枢神经系统的神经宿主防御机制中很重要。