Murai M, Aramaki Y, Tsuchiya S
School of Pharmacy, Tokyo University of Pharmacy and Life Science, Hachioji, Japan.
Immunology. 1995 Sep;86(1):64-70.
Culture supernatant, prepared by incubating fetal calf serum and liposome-treated B cells, augments mouse peritoneal macrophage Fc gamma receptor-mediated phagocytosis of IgG-opsonized sheep red blood cells. The activation process was hypothesized to be as follows. B-cell membranous glycosidases stimulated by liposomes, convert serum factor to a macrophage-stimulating active serum factor. As the active serum factor loses its activation potential by the addition of mannose or by digestion with alpha-mannosidase, mannose residues at the terminal present in the active serum factor are hypothesized to contribute to macrophage activation. The active serum factor was purified on a concanavalin A-Sepharose 4B column, and identified as a modified form of alpha 2-macroglobulin by immunochemical analysises. On non-denaturing polyacrylamide gel electrophoresis, modified alpha 2-macroglobulin showed a slow form, while alpha 2-macroglobulin-trypsin and alpha 2-macroglobulin-methylamine complexes, which bind specifically to alpha 2-macroglobulin receptors, showed a fast form and did not activate macropahges. These findings demonstrate that alpha 2-macroglobulin is the essential serum factor in liposome-primed macrophage activation, and that modified alpha 2-macroglobulin with mannose residues at the terminal sugar chain binds to macrophage mannose receptors, but not alpha 2-macroglobulin receptors, and increases Fc gamma receptor-mediated phagocytosis of IgG-opsonized sheep red blood cells.
通过将胎牛血清与脂质体处理的B细胞孵育制备的培养上清液,可增强小鼠腹腔巨噬细胞Fcγ受体介导的对IgG调理的绵羊红细胞的吞噬作用。激活过程被假设如下。脂质体刺激B细胞膜糖苷酶,将血清因子转化为巨噬细胞刺激活性血清因子。由于活性血清因子通过添加甘露糖或用α-甘露糖苷酶消化而失去其激活潜力,因此假设活性血清因子末端存在的甘露糖残基有助于巨噬细胞激活。活性血清因子在伴刀豆球蛋白A-Sepharose 4B柱上纯化,并通过免疫化学分析鉴定为α2-巨球蛋白的修饰形式。在非变性聚丙烯酰胺凝胶电泳上,修饰的α2-巨球蛋白显示为慢迁移形式,而特异性结合α2-巨球蛋白受体的α2-巨球蛋白-胰蛋白酶和α2-巨球蛋白-甲胺复合物显示为快迁移形式且不激活巨噬细胞。这些发现表明,α2-巨球蛋白是脂质体引发的巨噬细胞激活中的必需血清因子,并且末端糖链带有甘露糖残基的修饰α2-巨球蛋白与巨噬细胞甘露糖受体结合,但不与α2-巨球蛋白受体结合,并增加Fcγ受体介导的对IgG调理的绵羊红细胞的吞噬作用。