Different behaviour of two distinct types of Fc gamma receptor on guinea-pig peritoneal macrophages during phagocytosis of soluble immune complexes: identification of an intracellular pool for one of the receptors.

Guinea-pig peritoneal macrophages express two distinct types of Fc receptor for IgG (Fc gamma R): one specific for IgG2 (Fc gamma 2R) and the other for both IgG1 and IgG2 (Fc gamma 1/gamma 2R). When we employed flow cytometry for an assay, we found that the amount of ovalbumin (OA) complex of homologous IgG2 antibody bound on the surface of macrophages rapidly decreased during the phagocytosis in the presence of an excessive amount of the complex. This reduced binding capacity of the cells was gradually restored by incubating the cells in the complex-free medium, which showed that the Fc gamma Rs are consumed during the phagocytosis and again expressed on the cell surface. Flow cytometry with monoclonal anti-Fc gamma 2R Fab' and anti-Fc gamma 1/gamma 2R Fab' revealed that only the Fc gamma R type bound to the immune complex was selectively internalized, whereas another Fc gamma R type unbound persisted on the cell surface during the reaction. In addition, the amount of Fc gamma 1/gamma 2R on the cell surface was found to increase to a greater extent than did that of Fc gamma 2R, when phagocytosis was terminated by the removal of the immune complex. This result suggests that Fc gamma 1/gamma 2R is recruited from some intracellular store. In fact, we were able to demonstrate the existence of the membrane-associated intracellular Fc gamma 1/gamma 2R pool that increases the binding capacity of anti-Fc gamma 1/gamma 2R F(ab')2 by treatment of macrophages with saponin, and by fractionation of homogenized macrophages by sucrose density gradient centrifugation. The different behaviour of these two Fc gamma R type, thus shown, may cause the relatively sustained phagocytosing activity mediated by Fc gamma 1/gamma 2R compared with that caused by Fc gamma 2R; the former continued at least up to 6 hr, while the latter ceased within 2 hr.