A new photometric microassay for the quantitation of macrophage Fc receptor function. In vitro enzyme-containing immune complexes clearance (EIC) assay.
A new photometric microassay for immune complex (IC) binding to macrophages was developed in a homologous system using glucose oxidase-anti-glucose oxidase complexes (GOAGO) as a model for IC clearance in vitro. Thioglycollate-elicited murine peritoneal cells were incubated with GOAGO solution and then cell-associated glucose oxidase activity was measured after the washing and solubilization of the cell membrane in a microtitre plate. GOAGO binding to macrophages was inhibited in the presence of either IgG or its Fc fragments in a dose-dependent manner, while yeast mannan or IgG Fab fragments had no effect. These results indicated that this binding occurred solely via the Fc receptors on the macrophages. The Fc receptors for GOAGO were eliminated by trypsin digestion of the cells. When the macrophages were cultured with LPS or TPA, GOAGO binding was enhanced compared to that of control, whereas carrageenan treatment suppressed GOAGO binding. The present results suggest that this assay may be of value in the measurement of IC clearance and for studying the expression of Fc receptors on macrophages.
Matsumoto
T
T
Oriental Medicine Research Center, Kitasato Institute, Tokyo, Japan.
Tanaka
M
M
Yamada
H
H
Cyong
J C
JC
D000818:Animals; D000936:Antigen-Antibody Complex / Q000378:metabolism*; D001667:Binding, Competitive; D005949:Glucose Oxidase / Q000276:immunology; D007700:Kinetics; D008264:Macrophages / Q000276:immunology*; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D011961:Receptors, Fc / Q000032:analysis*; D013864:Thioglycolates; D014357:Trypsin
D000936:Antigen-Antibody Complex; D011961:Receptors, Fc; D013864:Thioglycolates; D005949:Glucose Oxidase; D014357:Trypsin