CELL-ADHESION MARKERS ARE EXPRESSED BY A STABLE HUMAN ENDOTHELIAL-CELL LINE TRANSFORMED BY THE SV40 LARGE T-ANTIGEN UNDER VIMENTIN PROMOTERCONTROL

Markers of endothelium have been studied in a new endothelial cell lin e derived from human umbilical cord vein cells by microinjection of a recombinant gene that includes a deletion mutant of the human vimentin gene regulatory region controlling the large T and small t antigen co ding region of the SV40 virus. In culture, this immortalized venous en dothelial cell line (IVEC) demonstrated morphological characteristics of endothelium; uptake of acetylated low density lipoprotein and prese nce of the Factor VIII-related antigen. Treatment of IVEC cells with I nterleukin-1beta (IL-1beta) at 10 U.ml-1 activates the expression of c ell adhesion molecules such as endothelial leucocyte adhesion molecule (ELAM-1), intercellular adhesion molecule-1 (ICAM-1), and vascular ce ll adhesion molecule-1 (VCAM-1), as observed in primary culture. Prost acyclin secretion was induced in the IVEC cells by 100 nM PMA treatmen t and thrombin at 0.5 U/ml. Angiotensin converting enzyme (ACE) activi ty detected in IVEC cells was present but lower than ACE activity in p rimary endothelial cells and was completely blocked by enalaprilat (1 muM), a specific ACE inhibitor. The presence of ACE mRNA was also demo nstrated in IVEC cells by RT-PCR amplification. Our data demonstrate t hat endothelial cells immortalized by use of this recombinant gene ret ain the morphological organization and numerous differentiated propert ies of endothelium. (C) 1993 Wiley-Liss, Inc.