Vitamin D3 binding protein required for in vitro activation of macrophages after alkylglycerol treatment of mouse peritoneal cells.
Vitamin D3 binding protein required for in vitro activation of macrophages after alkylglycerol treatment of mouse peritoneal cells.
复制标题
烷基甘油处理小鼠腹膜细胞后，体外激活巨噬细胞所需的维生素 D3 结合蛋白。
作者：
N. Yamamoto;S. Homma;J. Haddad;M. A. Kowalski
在10%胎牛血清（FCS）补充的培养基RPMI-1640中，用十二烷基甘油（DDG）体外处理腹膜细胞，导致巨噬细胞Fc受体介导的吞噬活性大大增强。这种巨噬细胞活化过程需要α 2-球蛋白组分中的血清因子。当小鼠腹腔细胞用50 ng/ml DDG在无血清的0.1%卵清蛋白补充培养基RPMI-1640（EA培养基）中处理30分钟，并在含有经离心分离的α_2-球蛋白的EA培养基中培养3小时时，观察到巨噬细胞的活化明显增强。为了提高α 2-球蛋白的分级分离，首先用50%饱和硫酸铵沉淀FCS，然后将上清液离心分级。所得的α 2-球蛋白组分不能支持巨噬细胞的活化。已知α 2-球蛋白级分的维生素D3结合蛋白（DBP）可被50%饱和硫酸铵沉淀。当用抗人DBP的抗血清处理人α 2-球蛋白并用于培养DDG处理的腹膜细胞的培养基中时，未观察到巨噬细胞的显著活化。在含有低浓度纯化的人DBP（组特异性组分，Gc）的培养基中培养DDG处理的腹膜细胞产生了大大增强的巨噬细胞的摄取活性。纯化的人Gc蛋白，当在EA培养基中用于DDG处理的B和未处理的T细胞的逐步培养时，被有效地转化为巨噬细胞活化因子。
In vitro treatment of peritoneal cells with dodecylglycerol (DDG) in 10% foetal calf serum (FCS) supplemented medium RPMI-1640 results in a greatly enhanced Fc receptor-mediated phagocytic activity of macrophages. This macrophage activation process requires a serum factor in the alpha 2-globulin fraction. When mouse peritoneal cells were treated with 50 ng DDG/ml in a serum-free 0.1% egg albumin-supplemented medium RPMI-1640 (EA medium) for 30 min and cultured in EA medium containing electrophoretically fractionated alpha 2-globulin for 3 hr, a markedly enhanced activation of macrophages was observed. To improve fractionation of alpha 2-globulin, FCS was first precipitated with 50% saturated ammonium sulphate and then the supernatant electrophoretically fractionated. The resultant alpha 2-globulin fraction was unable to support activation of macrophages. Vitamin D3 binding protein (DBP) of the alpha 2-globulin fraction is known to be precipitable by 50% saturated ammonium sulphate. When human alpha 2-globulin was treated with antiserum against human DBP and used in a medium for cultivation of DDG-treated peritoneal cells, no significant activation of macrophages was observed. Cultivation of DDG-treated peritoneal cells in a medium containing a low concentration of purified human DBP (group specific component, Gc) produced a greatly enhanced ingestion activity of macrophages. Purified human Gc protein, when used in an EA medium for step-wise cultivation of DDG-treated B and untreated T cells, was efficiently converted to a macrophage-activating factor.