Vitamin D-binding protein (group-specific component) is the sole serum protein required for macrophage activation after treatment of peritoneal cells with lysophosphatidylcholine
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S Homma1, M Yamamoto, N Yamamoto
11st Department of Internal Medicine, Jikei University, Tokyo, Japan.
Abstract:
In vitro treatment of mouse peritoneal cells with 1 micrograms lysophosphatidylcholine (lyso-Pc)/mL in serum free-0.1% egg albumin-supplemented RPMI 1640 medium for 30 min, followed by 3 h cultivation in a medium supplemented with human serum, resulted in a greatly enhanced Fc-receptor-mediated phagocytic activity of macrophages. Vitamin D-binding protein (group-specific component [Gc]) of alpha 2-globulin fraction was shown to be the sole serum glycoprotein required for the generation of a potent macrophage-activating factor. When a mixture of lysophosphatidylcholine (lyso-Pc)-treated nonadherent and adherent cells were cultured in a medium supplemented with a small amount of purified Gc protein (1 ng/mL), a greatly enhanced activation of macrophages was demonstrated. The generation of macrophage-activating factor from purified Gc protein was far more efficient than that from whole serum, indicating that a serum component is inhibitory to the activation process of macrophages. While three other major serum glycoproteins (alpha 2-macroglobulin, alpha 2-HS-glycoprotein and haptoglobin) were neither stimulatory nor inhibitory to lyso-Pc-primed macrophage activation, serum albumin (competitively with Gc protein) appeared to be inhibitory to the process of macrophage activation.
Insights
Lysophosphatidylcholine (lyso-Pc) treatment enhances macrophage phagocytosis. Vitamin D-binding protein (Gc) is key for activating macrophages, with purified Gc protein proving more effective than whole serum.
Area of Science:
- Immunology
- Cell Biology
Background:
- Macrophages play a crucial role in the immune system.
- Fc-receptor-mediated phagocytosis is a key function of macrophages.
Purpose of the Study:
- To investigate the in vitro activation of mouse macrophages.
- To identify serum components involved in macrophage activation.
Main Methods:
- Mouse peritoneal cells were treated with lysophosphatidylcholine (lyso-Pc).
- Cells were cultured in media supplemented with human serum or purified glycoproteins.
- Fc-receptor-mediated phagocytic activity was measured.
Main Results:
- Lyso-Pc treatment significantly enhanced macrophage Fc-receptor-mediated phagocytic activity.
- Vitamin D-binding protein (Gc) was identified as the sole essential serum glycoprotein for generating macrophage-activating factor.
- Purified Gc protein was more efficient in activating macrophages than whole serum.
- Serum albumin inhibited Gc protein-mediated macrophage activation.
Conclusions:
- Vitamin D-binding protein (Gc) is critical for lysophosphatidylcholine-induced macrophage activation.
- A serum component, likely albumin, inhibits this activation process.
- Optimized conditions using purified Gc protein can enhance macrophage activation.