Human monoclonal IgG isotypes differ in complement activating function at the level of C4 as well as C1q.
Humanized antibodies are likely to have a major role in therapy and it is important to define their interaction with physiological effectors. By comparing a matched series of chimeric human mAbs we found that igG1 was most efficient in complement lysis, although IgG3 bound more C1q. To resolve this paradox we compared the ability of human IgG1, IgG2, IgG3, IgG4, and IgE and rat IgG2b to cause C1q binding, C1 binding and activation, C4 activation, C4b binding, and C3b binding. Rat IgG2b was included because this isotype has already successfully been used for therapy. Human IgG1 was less efficient than IgG3 and fixing C1q and C1 on the cell surface, but the number of C4 molecules bound per C1 was 10-fold greater for IgG1 than for IgG3. This difference, amplified through later stages of the complement cascade, can account for the superiority of IgG1 for cell lysis. The efficiency of IgG1 in fixing C4 was not due to a favored binding site on the antibody molecule, since virtually all of the bound C4b was attached to the cells. Rather, it appeared that the activation of C4 by C1s was greatly favored by IgG1 compared with IgG3. It should be possible to combine the optimal properties of IgG1 and IgG3 antibodies to produce an improved therapeutic reagent.
Bindon
C I
CI
Department of Pathology, University of Cambridge, United Kingdom.
Hale
G
G
Brüggemann
M
M
Waldmann
H
H
D000911:Antibodies, Monoclonal / Q000276:immunology*; D003166:Complement Activating Enzymes / Q000276:immunology*; D003167:Complement Activation; D003172:Complement C1 / Q000276:immunology*; D015922:Complement C1q; D003176:Complement C3 / Q000276:immunology; D003179:Complement C3b / Q000276:immunology; D003181:Complement C4 / Q000276:immunology*; D015935:Complement C4b; D004910:Erythrocyte Membrane / Q000276:immunology; D004912:Erythrocytes / Q000276:immunology; D006241:Haptens; D006461:Hemolysis; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology*; D007132:Immunoglobulin Isotypes / Q000276:immunology; D009592:Nitrohydroxyiodophenylacetate / Q000276:immunology
D000911:Antibodies, Monoclonal; D003172:Complement C1; D003176:Complement C3; D003181:Complement C4; D006241:Haptens; D007074:Immunoglobulin G; D007132:Immunoglobulin Isotypes; D009592:Nitrohydroxyiodophenylacetate; D015922:Complement C1q; D003179:Complement C3b; D015935:Complement C4b; D003166:Complement Activating Enzymes