Mapping studies reveal unique epitopes on IgG recognized by rheumatoid arthritis-derived monoclonal rheumatoid factors.
We have used chimeric IgG antibodies and their genetically engineered variants prepared by a combination of site-directed mutagenesis and exon exchange to define the structure(s) on IgG recognized by monoclonal rheumatoid factor (RF) autoantibodies from rheumatoid arthritis (RA) patients. Nineteen RF produced by EBV-transformed cell lines from the synovium or blood of RA patients were analyzed. Their binding patterns differ significantly from those seen with RF obtained from patients with Waldenstrom's macroglobulinemia (WMac). Half of the RA-derived RF bound IgG1, 2, and 4, but not 3 (Ga specificity), the common pattern in WMac. However, heterogeneity in fine specificity within the Ga reactivity pattern was observed. Moreover, seven others bound all four IgG subclasses, a pattern observed for only one WMac-derived RF from a patient who also had RA. Three RF had subclass specificities unlike any observed with WMac-derived RF. Most RA-derived RF bound IgG at a discontinuous epitope comprised of residues from both the CH2 and CH3 H chain constant regions. However, unlike any WMac-derived RF, one RA-derived RF bound IgG in CH2, another in CH3, and a third at an undetermined site outside of the CH2-CH3 interface. Some RA-derived RF bound aglycosylated IgG4 less well than glycosylated IgG4, suggesting that the carbohydrate moiety was important in establishing their binding epitope in CH2. These studies demonstrate that the repertoire of RF expressed by RA patients contains some unique binding specificities for IgG epitopes not found among our panel of WMac-derived RF. Our results therefore call into question whether WMac-derived RF with their limited diversity are appropriate models for disease-related RF. In addition, RF with their multiple specificities can serve as probes of antibody structure.
Bonagura
V R
VR
Department of Pediatrics, Schneider Children's Hospital of Long Island Jewish Medical Center, New Hyde Park, NY 11042.
Artandi
S E
SE
Davidson
A
A
Randen
I
I
Agostino
N
N
Thompson
K
K
Natvig
J B
JB
Morrison
S L
SL
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D001172:Arthritis, Rheumatoid / Q000276:immunology*; D002478:Cells, Cultured; D000939:Epitopes / Q000032:analysis*; D006031:Glycosylation; D006801:Humans; D007074:Immunoglobulin G / Q000145:classification / Q000235:genetics / Q000276:immunology*; D007132:Immunoglobulin Isotypes / Q000276:immunology; D051379:Mice; D016297:Mutagenesis, Site-Directed; D011993:Recombinant Fusion Proteins / Q000276:immunology; D012217:Rheumatoid Factor / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000939:Epitopes; D007074:Immunoglobulin G; D007132:Immunoglobulin Isotypes; D011993:Recombinant Fusion Proteins; D012217:Rheumatoid Factor