The p68 autoantigen characteristic of rheumatoid arthritis is reactive with carbohydrate epitope specific autoantibodies.
OBJECTIVE
The autoantigen p68 is a target of autoantibodies as well as autoreactive T cells with a high specificity in rheumatoid arthritis (RA). The binding characteristics of the autoantibodies to their antigen were now analysed biochemically and cytologically.


METHODS
Deglycosylation techniques as well as lectin and sugar competition experiments were performed to p68 to discover if the antibodies detected a glycoepitope, Its antigenicity was investigated applying anti-p68 antibodies derived from RA patients in comparison with polyclonal rabbit anti-p68 antibodies.


RESULTS
p68 specific antibodies from RA patients did not to bind to p68 that had been deglycosylated by alkaline beta-elimination, O-glycosidase or periodate treatment. In contrast, binding of p68 specific antibodies raised in rabbit was unaffected by either deglycosylation protocol. Furthermore, lectins specific for the carbohydrate N-acetylglucosamine competed with p68 specific antibodies from RA patients for antigen bindings. N-acetylglucosamine by itself also competed with patient derived anti-p68 antibodies for p68 binding. Again, rabbit and anti-p68 antibodies did not elicit these competitive effects. Applying cytoimmunofluorescence, p68 was present in the cytoplasm or endoplasmic reticulum and also in low abundance on the cell surface. Under heatshock conditions, p68 was detectable in the nucleus.


CONCLUSIONS
Autoimmunity to p68 during RA is carried by anti-carbohydrate autoantibodies. The carbohydrate modification of p68 appears to be N-acetylglucosamine, which may reflect the regulation of intracellular localisation of the antigen. It is hypothesised that a shift in glycosylation pattern accompanied by an unphysiological localisation of the antigen could trigger antigenicity of p68 during the pathogenesis of IRA.
Department of Medicine III, Rheumatology, Charité University Hospital, Humboldt University of Berlin, Germany.
S
S
Bläss
C
C
Meier
H W
HW
Vohr
M
M
Schwochau
C
C
Specker
G R
GR
Burmester
D001172:Arthritis, Rheumatoid / Q000276:immunology*; D001323:Autoantibodies / Q000378:metabolism*; D001324:Autoantigens / Q000276:immunology*; D001667:Binding, Competitive; D015153:Blotting, Western; D002241:Carbohydrates / Q000276:immunology*; D000939:Epitopes; D006031:Glycosylation; D006367:HeLa Cells; D006801:Humans; D037102:Lectins / Q000378:metabolism; D008856:Microscopy, Fluorescence; D011485:Protein Binding
D001323:Autoantibodies; D001324:Autoantigens; D002241:Carbohydrates; D000939:Epitopes; D037102:Lectins