Molecular cloning and expression of 56-58 KD antigen associated with transplant coronary artery disease.
High circulating levels of anti-endothelial antibodies have been reported in patients with accelerated transplant coronary artery disease (TxCAD); however, the nature or characteristics of the antigen to which these antibodies bind in vivo remain unclear. To identify the antigen, a human endothelial cell cDNA expression library was constructed in lambdaZAP and immuno-screened with serum containing a high titre of anti-endothelial antibodies. Nucleotide sequence of cloned cDNA inserts and GenBank database searches revealed a 94% identity in a 35 bp overlap to the human vimentin gene. The cloned antigen fusion protein co-migrated with human vimentin of molecular mass 56-58 KD in SDS-PAGE and exhibited immunoreactivity towards monoclonal and polyclonal anti-human vimentin antibodies. These results suggest that the 56-58 KD antigen associated with accelerated TxCAD is vimentin (or a vimentin-like protein), a cytoskeletal protein present in cells of the blood vessel walls.
Ationu
A
A
Department of Haematological Medicine, Kings College School of Medicine and Dentistry, London, United Kingdom. aationu@hgmp.mrc.ac.uk
Collins
A
A
D000911:Antibodies, Monoclonal; D001323:Autoantibodies / Q000097:blood; D001324:Autoantigens / Q000235:genetics*; D001483:Base Sequence; D015153:Blotting, Western; D003001:Cloning, Molecular; D003327:Coronary Disease / Q000209:etiology / Q000276:immunology*; D015180:Electrophoresis, Gel, Two-Dimensional; D004730:Endothelium, Vascular / Q000276:immunology*; D015870:Gene Expression; D016027:Heart Transplantation / Q000009:adverse effects*; D006801:Humans; D008969:Molecular Sequence Data; D017385:Sequence Homology; D014746:Vimentin / Q000235:genetics
D000911:Antibodies, Monoclonal; D001323:Autoantibodies; D001324:Autoantigens; D014746:Vimentin