Monoclonal antibodies that bind to distinct epitopes on Fc gamma RI are able to trigger receptor function.
The human monocyte and macrophage Fc receptor that binds human IgG with high affinity is a surface glycoprotein with a relative molecular mass of approximately 70 kDa. This receptor (Fc gamma RI) has been partially characterized using mAb 32 which binds outside the Fc binding domain of the receptor, but nonetheless triggers Fc receptor-dependent functions. In this study, we describe the properties of four new antibodies with specificity for Fc gamma RI. Based on additivity and cross-blocking studies, we conclude that two of these antibodies (mAb 22 and 44) define a third epitope which is distinct from the binding sites for both mAb 32 and the Fc portion of human IgG. Each Fc gamma RI-specific hybridoma was selected for stable sublines expressing high levels of mAb on the cell surface, and then tested for the ability of this surface mAb to trigger antibody-dependent cell cytotoxicity. All sublines were killed by human monocytes when used as targets in a 51Cr-release assay, whereas hybridomas specific for myeloid Ag other than Fc gamma RI were not killed. We conclude that Fc receptor function is triggered through binding to each of the three epitopes of Fc gamma RI that we have defined. These mAb will be useful for additional characterization of Fc gamma RI, and may, when incorporated into tumor-directed heteroantibodies, enhance tumor cell killing by human monocytes and macrophages.
Guyre
P M
PM
Department of Physiology, Dartmouth Medical School, Hanover, NH 03756.
Graziano
R F
RF
Vance
B A
BA
Morganelli
P M
PM
Fanger
M W
MW
D000818:Animals; D000911:Antibodies, Monoclonal / Q000502:physiology*; D000915:Antibody Affinity; D000918:Antibody Specificity; D000943:Antigens, Differentiation / Q000276:immunology / Q000378:metabolism* / Q000502:physiology; D001666:Binding Sites, Antibody; D002460:Cell Line; D003602:Cytotoxicity, Immunologic; D000939:Epitopes / Q000032:analysis / Q000276:immunology*; D016650:Fluorescein-5-isothiocyanate; D005452:Fluoresceins; D006801:Humans; D007074:Immunoglobulin G / Q000378:metabolism*; D051379:Mice; D011961:Receptors, Fc / Q000276:immunology / Q000378:metabolism* / Q000502:physiology; D017452:Receptors, IgG; D013861:Thiocyanates
D000911:Antibodies, Monoclonal; D000943:Antigens, Differentiation; D000939:Epitopes; D005452:Fluoresceins; D007074:Immunoglobulin G; D011961:Receptors, Fc; D017452:Receptors, IgG; D013861:Thiocyanates; D016650:Fluorescein-5-isothiocyanate