Activation of effector functions by immune complexes of mouse IgG2a with isotype-specific autoantibodies.
Analysis of five monoclonal autoantibodies, rheumatoid factors produced by hybridomas generated from spleen cells of BALB/c mice repeatedly infected with A/PR/8/34 human influenza A virus, revealed that they recognized distinct but spatially related epitopes. The differing isoallotypic specificity of the IgM and IgA monoclonal antibodies correlated with the presence of Ile258 and Ala305, respectively. Although these data suggest that the epitopes recognized are within the CH2 domain, all antibodies failed to inhibit IgG antigen reactivity with Staphylococcus aureus protein A (SpA), C1q, mouse C3, human Fc gamma RI or mouse Fc gamma RII, activities known to be predominantly determined by CH2 domain structures. Reactivity of the IgA antibody, Z34, with IgG2b allowed further specificity studies using a panel of 26 mutant IgG2b proteins, each having single amino acid replacements over the surface of the CH2 domain. The only substitution that affected Z34 reactivity was Asn/Ala297, which destroyed the glycosylation sequon, resulting in secretion of an aglycosylated IgG molecule. The epitope recognized by Z34 therefore seems to be located outside of the Fc gamma R and C1q binding sites, but to be dependent on the presence of carbohydrate for expression. In contrast to the binding studies, complement activation by aggregated IgG2a, through classical or alternative pathways, was inhibited by the presence of autoantibodies. The functional significance of isotype-specific autoantibody in immune regulation is discussed.
Rajnavölgyi
E
E
Department of Immunology, L. Eötvös University, Göd, Hungary.
Fazekas
G
G
Lund
J
J
Daeron
M
M
Teillaud
J L
JL
Jefferis
R
R
Fridman
W H
WH
Gergely
J
J
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000918:Antibody Specificity / Q000276:immunology*; D000936:Antigen-Antibody Complex / Q000276:immunology*; D003167:Complement Activation / Q000276:immunology; D015922:Complement C1q / Q000378:metabolism; D003176:Complement C3 / Q000378:metabolism; D000939:Epitopes / Q000276:immunology; D007070:Immunoglobulin A / Q000276:immunology; D007074:Immunoglobulin G / Q000737:chemistry / Q000276:immunology*; D007132:Immunoglobulin Isotypes / Q000276:immunology*; D007075:Immunoglobulin M / Q000276:immunology; D051379:Mice; D017452:Receptors, IgG / Q000276:immunology; D012217:Rheumatoid Factor / Q000276:immunology*; D013205:Staphylococcal Protein A / Q000276:immunology
D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D003176:Complement C3; D000939:Epitopes; D007070:Immunoglobulin A; D007074:Immunoglobulin G; D007132:Immunoglobulin Isotypes; D007075:Immunoglobulin M; D017452:Receptors, IgG; D013205:Staphylococcal Protein A; D015922:Complement C1q; D012217:Rheumatoid Factor