Identification of the serum factor required for in vitro activation of macrophages. Role of vitamin D3-binding protein (group specific component, Gc) in lysophospholipid activation of mouse peritoneal macrophages.
Identification of the serum factor required for in vitro activation of macrophages. Role of vitamin D3-binding protein (group specific component, Gc) in lysophospholipid activation of mouse peritoneal macrophages.
复制标题
鉴定巨噬细胞体外激活所需的血清因子。
DOI：
10.4049/jimmunol.147.1.273
复制
发表时间：
1991
影响因子：
4.4
通讯作者：
I. Millman
中科院分区：
文献类型：
--
作者：
N. Yamamoto;S. Homma;I. Millman
在体外处理小鼠腹膜细胞（粘附和非粘附细胞的混合物）与溶血磷脂酰胆碱（lyso-Pc）在10%FCS补充培养基RPMI 1640的结果大大增强FcR介导的巨噬细胞的吞噬活性。这种巨噬细胞活化过程需要血清因子。胎牛和人血清的淀粉块电泳分级研究表明，α 2-球蛋白组分含有巨噬细胞活化所必需的血清因子。为了鉴定血清因子，人血清用50%饱和硫酸铵沉淀，并在SephadexG-100柱上分级。具有较低m.w.比白蛋白具有支持巨噬细胞活化的能力。使用抗人α 2-球蛋白的三种主要蛋白的兔抗血清，通过免疫吸收分析该蛋白组分中的活性血清因子。这种活性血清因子被证明是一种维生素D3结合蛋白（组特异性组分，Gc）。通过使用人血清的单克隆抗Gc吸收的活性柱部分，我们观察到没有增强的巨噬细胞活化超过溶血-Pc处理的腹膜细胞的无血清部分培养的结果。在含有低浓度纯化的人Gc蛋白（0.1至2.6 ng/ml）的培养基中培养lyso-Pc处理的腹膜细胞产生了大大增强的巨噬细胞吞噬活性。当纯化的人Gc蛋白被用于在无血清培养基中逐步培养溶血-Pc处理的非粘附细胞类型时，有效地产生了巨噬细胞活化因子。因此，可以得出结论，维生素D3结合蛋白是溶血Pc引发的巨噬细胞活化的必要血清因子。
In vitro treatment of mouse peritoneal cells (mixture of adherent and nonadherent cells) with lysophosphatidylcholine (lyso-Pc) in 10% FCS supplemented medium RPMI 1640 results in a greatly enhanced FcR-mediated phagocytic activity of macrophages. This macrophage-activation process requires a serum factor. Fractionation studies with starch block electrophoresis of fetal calf and human sera revealed that alpha 2-globulin fraction contains a serum factor essential for macrophage activation. To identify the serum factor, human serum was precipitated with 50% saturated ammonium sulfate and fractionated on a Sephadex G-100 column. A protein fraction with a lower m.w. than albumin had the capacity to support activation of macrophages. The active serum factor in this protein fraction was analyzed by immunoabsorption by using rabbit antisera against three major proteins of human alpha 2-globulin. This active serum factor was shown to be a vitamin D3-binding protein (group specific component, Gc). By using a monoclonal anti-Gc-absorbed active column fraction of human serum, we observed no enhanced macrophage activation over the results with serum fraction-free cultivation of lyso-Pc-treated peritoneal cells. Cultivation of lyso-Pc-treated peritoneal cells in a medium containing a low concentration of purified human Gc protein (0.1 to 2.6 ng/ml) produced a greatly enhanced phagocytic activity of macrophages. When purified human Gc protein was used in a serum-free medium for stepwise cultivation of lyso-Pc-treated nonadherent cell types, a macrophage-activating factor was efficiently generated. Therefore, it is concluded that the vitamin D3-binding protein is the essential serum factor for the lyso-Pc-primed activation of macrophages.