Differential binding to human FcgammaRIIa and FcgammaRIIb receptors by human IgG wildtype and mutant antibodies.
We are investigating the interactions of recombinant human IgG antibodies with Fc receptors to enable selection of a constant region giving minimal depletion of antigen-bearing cells. Eight variant constant regions were made by substituting motifs between human IgG subclasses in the lower hinge region and/or a specially close loop of the CH2 domain. Mutations in the lower hinge region were shown to eliminate FcgammaRI binding and monocyte activation [Eur. J. Immunol. 29 (1999) 2613]. Here, we detail interactions with FcgammaRIIa of the 131R and 131H allotypes and FcgammaRIIb. Lower hinge mutations caused large reductions in binding whereas modification of residues 327, 330 and 331 had less dramatic effects. However, like the wildtype IgG subclass binding hierarchies, the effect of the mutations varied between different receptors. We identified IgG1 variants which react with the activating receptor, FcgammaRIIa, at least 10-fold less efficiently than wildtype IgG1 but whose binding to the inhibitory receptor, FcgammaRIIb, is only four-fold reduced. Manipulation of interactions with FcgammaRIIb separately from those with activating receptors provides potential for designing antibodies with novel and effective combinations of attributes. In addition, insight is gained into the evolution of functional differences in human IgG subclasses.
Armour
Kathryn L
KL
Department of Pathology, Division of Immunology, University of Cambridge, Tennis Court Road, Cambridge CB2 1QP, UK.
van de Winkel
Jan G J
JG
Williamson
Lorna M
LM
Clark
Mike R
MR
D000906:Antibodies / Q000276:immunology*; D015703:Antigens, CD / Q000276:immunology*; D001666:Binding Sites, Antibody; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology*; D008958:Models, Molecular; D009154:Mutation; D017452:Receptors, IgG / Q000276:immunology*
D000906:Antibodies; D015703:Antigens, CD; C080568:Fc gamma receptor IIA; C080571:Fc gamma receptor IIB; D007074:Immunoglobulin G; D017452:Receptors, IgG