The identification of endothelial cell autoantigens.
The identity of many endothelial cell autoantigens remains unclear. This study has used human monoclonal anti-endothelial cell autoantibodies isolated from patients with SLE to identify endothelial autoantigens. Thirteen antibodies reactive with endothelial cell membrane preparations were isolated and cloned, one of which has previously been demonstrated to be pro-inflammatory. Western blotting demonstrates that these antibodies recognize a variety of proteins in endothelial cell membrane preparations. Further characterization of five antibodies by cDNA library screening, immunofluorescence and Western blotting proves that two of these antibodies recognized the cytoskeletal proteins tubulin and vimentin. A further antibody identified a clone derived from human collagenase, an identification supported by Western blotting. The multiple clones selected by other antibodies are not compatible with the molecular weight of the antigen recognized in Western blotting studies. This study has clearly identified two endothelial cell autoantigens present in membrane preparations and provides strong evidence as to the identity of a third.
Yazici
Z A
ZA
Department of Biomedical Sciences, University of Bradford, Bradford, West Yorkshire, BD7 1DP, UK.
Behrendt
M
M
Cooper
D
D
Goodfield
M
M
Partridge
L
L
Lindsey
N J
NJ
D000911:Antibodies, Monoclonal / Q000378:metabolism; D000937:Antigen-Antibody Reactions; D001324:Autoantigens / Q000032:analysis* / Q000276:immunology; D015153:Blotting, Western; D002460:Cell Line; D018076:DNA, Complementary / Q000032:analysis; D004730:Endothelium, Vascular / Q000737:chemistry* / Q000166:cytology / Q000276:immunology*; D019084:Fluorescent Antibody Technique, Indirect; D015723:Gene Library; D006801:Humans; D020014:K562 Cells; D008970:Molecular Weight
D000911:Antibodies, Monoclonal; D001324:Autoantigens; D018076:DNA, Complementary