Characterization of the Fc receptor for IgG2 on guinea pig polymorphonuclear leukocytes by the use of a monoclonal antibody.
Guinea pig polymorphonuclear leukocytes (PMNs) possess two distinct types of Fc gamma receptor (Fc gamma R): Fc gamma 1/gamma 2R for both IgG1 and IgG2, and Fc gamma 2R for IgG2 alone. The Fc gamma 2R was previously shown to differ antigenically from homologous macrophage (M phi) Fc gamma 2R by the use of a monoclonal antibody to M phi Fc gamma 2R (VIIAI IgG1), though the Fc gamma 1/gamma 2R cross-reacts with a monoclonal antibody to homologous M phi Fc gamma 1/gamma 2R (VIA2 IgG1). Recently, we obtained a monoclonal antibody (MP-2) secreted by a hybridoma prepared by fusion of the splenic cells of mice immunized with guinea pig PMNs with a myeloma cell line. This antibody completely inhibited both the Fc gamma 2R-mediated rosette formation of PMNs with IgG2 antibody-sensitized sheep erythrocytes and the Fc gamma 2R-mediated binding of ovalbumin (OA)-complexed IgG2 antibody to PMNs. When the antigen of MP-2 was isolated by affinity chromatography with the antibody-Sepharose, it gave a single band with a molecular weight of 120,000 on SDS-PAGE. The number of antigen molecules per PMN was estimated to be 9 X 10(4) by measuring the binding of 125I-MP-2 Fab. This value was essentially the same as that obtained by measuring the binding of OA-complexed IgG2 antibody to the PMNs treated with the Fab' of VIA2 IgG1. These results strongly suggest that MP-2 is a monoclonal antibody to PMN Fc gamma 2R.
Sato
M
M
Department of Hygienic Chemistry, Faculty of Pharmaceutical Sciences, Hokkaido University.
Koyama
J
J
D000818:Animals; D000911:Antibodies, Monoclonal; D000937:Antigen-Antibody Reactions; D004591:Electrophoresis, Polyacrylamide Gel; D004912:Erythrocytes / Q000276:immunology; D006168:Guinea Pigs; D007074:Immunoglobulin G / Q000276:immunology*; D066298:In Vitro Techniques; D007457:Iodine Radioisotopes; D051379:Mice; D008970:Molecular Weight; D009504:Neutrophils / Q000276:immunology*; D011946:Receptors, Antigen / Q000276:immunology*; D011961:Receptors, Fc / Q000032:analysis*; D012397:Rosette Formation; D012756:Sheep / Q000276:immunology
D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D007457:Iodine Radioisotopes; D011946:Receptors, Antigen; D011961:Receptors, Fc