Human IgG1 and its Fc fragment bind with different affinities to the Fc receptors on the human U937, HL-60 and ML-1 cell lines.
Measurements were made of the binding of human monomeric 125I-IgG1 and 125I-Fc to U937 cells at room temp. Analyses of the binding data showed that these cells possessed a single class of receptor (FcR) for Fc or IgG and, although both ligands were found to bind to the same number of sites per cell, Fc was found to bind with about twice the affinity of IgG. At 20 degrees C estimates of the forward rate constants and the dissociation rate constants for IgG and Fe were 1.13 and 3.65 X 10(7) M-1 min-1 and 0.33 and 0.57 X 10(-2) min-1 respectively. Independent determinations of the association constants (Ka) under the same experimental conditions gave values of 0.98 X 10(9) M-1 for IgG and 3.1 X 10(9) M-1 for Fc. Thus the Fc fragment of IgG appears to bind to U937 FcR at 3-4 times the rate of IgG and to dissociate at about twice the rate, resulting in higher values of Ka for the Fc-FcR than for the IgG-FcR interaction. Also, in competitive-binding experiments and in EA rosette inhibition assay the Fc fragment was consistently found to be more efficient in FcR binding than IgG. Similar results were obtained using HL-60 and ML-1 cells which possess FcR like those on U937 cells and with IgG1 and Fc prepared from other myelomas. IgG and Fe which had undergone mild reduction and alkylation bound to the same number of FcR per U937 cell as the non-reduced ligands but the affinity of binding was diminished to a similar degree with both ligands, suggesting that the major effect of cleavage of the interchain disulfide bonds on cytophilic binding is due to alteration of the native quaternary relationships of the C gamma 2 and C gamma 3 domains.
D000915:Antibody Affinity; D001667:Binding, Competitive; D002460:Cell Line; D006801:Humans; D007141:Immunoglobulin Fc Fragments / Q000276:immunology*; D007128:Immunoglobulin Fragments / Q000276:immunology*; D007074:Immunoglobulin G / Q000276:immunology*; D007457:Iodine Radioisotopes; D007700:Kinetics; D011961:Receptors, Fc / Q000276:immunology*; D012397:Rosette Formation
D007141:Immunoglobulin Fc Fragments; D007128:Immunoglobulin Fragments; D007074:Immunoglobulin G; D007457:Iodine Radioisotopes; D011961:Receptors, Fc