An SV40 large T-antigen immortalized human umbilical vein endothelial cell line for anti-endothelial cell antibody detection.
OBJECTIVE
Anti-endothelial cell antibodies in serum of patients with different inflammatory diseases can be detected by a whole cell enzyme-linked immunosorbant assay, using primary cultures of human umbilical vein endothelial cells. To avoid repeated isolation, it would be of great value if an immortal endothelial cell line could be used to perform anti-endothelial cell antibody assays.


METHODS
In this study endothelial cells from human umbilical and iliac veins and arteries were transfected with a plasmid containing the Simian Virus 40 large T-antigen. Endothelial cell line(s) derived from this procedure were compared with human umbilical vein endothelial cells in the anti-endothelial cell antibody assay.


RESULTS
After transfection, clones of homologous cell populations showed an extended lifespan, before entering a period of crisis. In one human umbilical 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 was a significant correlation between anti-endothelial cell antibody testing on human umbilical vein endothelial cells and on the cell line EVLC2.


CONCLUSIONS
The Simian Virus 40 large T-antigen immortalized human umbilical vein endothelial cell line EVLC2 may be useful for the detection of anti-endothelial cell antibodies.
van Leeuwen
E B
EB
Groningen University Institute for Drug Exploration (GUIDE): Department of Haematology, Division of Haemostasis, Thrombosis and Rheology, University of Groningen and University Hospital Groningen, Groningen, The Netherlands. e.vanleeuwen@immuno.unimaas.nl
Wisman
G B
GB
Tervaert
J W
JW
Palmans
L L
LL
van Wijk
R T
RT
Veenstra
R
R
Molema
G
G
van der Zee
A G
AG
van der Meer
J
J
Ruiters
M H
MH
D000952:Antigens, Polyomavirus Transforming / Q000235:genetics*; D001323:Autoantibodies / Q000032:analysis* / Q000276:immunology*; D002461:Cell Line, Transformed; D004730:Endothelium, Vascular / Q000166:cytology* / Q000201:enzymology / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D014890:Granulomatosis with Polyangiitis / Q000175:diagnosis / Q000276:immunology*; D006801:Humans; D015850:Interleukin-6 / Q000032:analysis; D016209:Interleukin-8 / Q000032:analysis; D053146:Myeloblastin; D009195:Peroxidase / Q000276:immunology; D012697:Serine Endopeptidases / Q000276:immunology; D019098:Telomerase / Q000378:metabolism; D016615:Telomere / Q000378:metabolism; D014162:Transfection; D014471:Umbilical Veins / Q000166:cytology
D000952:Antigens, Polyomavirus Transforming; D001323:Autoantibodies; D015850:Interleukin-6; D016209:Interleukin-8; C087589:anti-endothelial cell antibody; D009195:Peroxidase; D019098:Telomerase; D012697:Serine Endopeptidases; D053146:Myeloblastin