An SV40 large T-antigen immortalized human umbilical vein endothelial cellline for anti-endothelial cell antibody detection

Objective Anti-endothelial cell antibodies in serum of patients with differ ent inflammatory diseases can be detected by, a whole cell enzyme-linked im munosorbant assay using primary cultures of human umbilical vein endothelia l cells. To avoid repeated isolation, it would be of great value if an immo rtal endothelial cell line could be used to perform anti-endothelial cell a ntibody assays. Methods In this study endothelial cells from human umbilical and iliac vein s and arteries were transfected with a plasmid containing the Simian Virus 40 large T-antigen. Endothelial cell line(s) derived from this procedure we re compared with human umbilical vein endothelial cells in the anti-endothe lial cell antibody assay. Results After transfection, clones of homologous cell populations showed an extended lifespan, before entering a period of crisis. In one human umbili cal vein endothelial cell clone a subpopulation of cells escaped crisis and became immortal (EVLC2). Telomerase was activated in this endothelial cell line, resulting in maintenance of the telomere length. There,vas a signifi cant correlation between anti-endothelial cell antibody testing on human um bilical vein endothelial cells and on the cell line EVLC2. Conclusion The Simian Virus 40 large T-antigen immortalized human umbilical vein endothelial cell line EVLC2 may be useful for the detection of anti-e ndothelial cell antibodies.