IgG isotype-specific auto-antibodies bind preferentially to cross-linked membrane Ig.
Under equilibrium conditions, the affinities of five anti-IgG2a mAb isolated from virus-infected mice were comparable to other high-affinity auto-antibodies. Similar to rheumatoid factors, these anti-IgG2a auto-antibodies bound to aggregated or complexed IgG2a with 50 to 1500-fold higher avidity than their monomeric counterparts. Despite their high functional affinity to IgG2a, flow cytometric analysis revealed no binding or marginal mAb binding to four distinct lines of B cells expressing different densities of membrane-anchored IgG2a. If, however, surface IgG2a was cross-linked by polyclonal light chain-specific antibodies, IgM and IgA mAb binding resulted, and was detected as an increase in mean fluorescence intensity compared with isotype-matched control antibodies. The binding of one IgM mAb to cross-linked IgG2a patches of the cell surface was also visualized by confocal microscopy. Pretreatment of cells with aggregated IgG2a caused increased fluorescence intensity, demonstrating that the IgM and IgA mAb were also able to interact with IgG2a aggregates bound on the B cell surface via Fc gamma RIIB. It also permitted efficient co-ligation of the aggregated B cell receptors (BCR) with Fc gamma RIIB-fixed immune complexes known to deliver a negative signal in B cell activation. Cross-linking of IgG2a complexes bound to Fc gamma RI on macrophages or dendritic cells with antigen-specific BCR and/or T cells via their Fc gamma RIIB may accelerate the physical contact of cells involved in the antigen-specific response.(ABSTRACT TRUNCATED AT 250 WORDS)
Sandoz Research Institute, Vienna, Austria.
G
G
Fazekas
G
G
Pálfi
B
B
Wolff-Winiski
B
B
Rosenwirth
P
P
Dukor
J
J
Gergely
E
E
Rajnavölgyi
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry; D000915:Antibody Affinity; D015703:Antigens, CD / Q000737:chemistry; D001323:Autoantibodies / Q000737:chemistry*; D001402:B-Lymphocytes / Q000737:chemistry; D001666:Binding Sites, Antibody; D002449:Cell Aggregation / Q000276:immunology; D002462:Cell Membrane / Q000737:chemistry / Q000276:immunology; D003432:Cross-Linking Reagents; D006801:Humans; D007074:Immunoglobulin G / Q000737:chemistry*; D007132:Immunoglobulin Isotypes / Q000737:chemistry*; D051379:Mice; D011947:Receptors, Antigen, B-Cell / Q000737:chemistry*; D017452:Receptors, IgG / Q000737:chemistry; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D001323:Autoantibodies; D003432:Cross-Linking Reagents; C080571:Fc gamma receptor IIB; D007074:Immunoglobulin G; D007132:Immunoglobulin Isotypes; D011947:Receptors, Antigen, B-Cell; D017452:Receptors, IgG