Cell adhesion markers are expressed by a stable human endothelial cell line transformed by the SV40 large T antigen under vimentin promoter control.
Markers of endothelium have been studied in a new endothelial cell line 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 coding region of the SV40 virus. In culture, this immortalized venous endothelial cell line (IVEC) demonstrated morphological characteristics of endothelium; uptake of acetylated low density lipoprotein and presence of the Factor VIII-related antigen. Treatment of IVEC cells with Interleukin-1 beta (IL-1 beta) at 10 U.ml-1 activates the expression of cell adhesion molecules such as endothelial leucocyte adhesion molecule (ELAM-1), intercellular adhesion molecule-1 (ICAM-1), and vascular cell adhesion molecule-1 (VCAM-1), as observed in primary culture. Prostacyclin secretion was induced in the IVEC cells by 100 nM PMA treatment and thrombin at 0.5 U/ml. Angiotensin converting enzyme (ACE) activity detected in IVEC cells was present but lower than ACE activity in primary endothelial cells and was completely blocked by enalaprilat (1 microM), a specific ACE inhibitor. The presence of ACE mRNA was also demonstrated in IVEC cells by RT-PCR amplification. Our data demonstrate that endothelial cells immortalized by use of this recombinant gene retain the morphological organization and numerous differentiated properties of endothelium.
Vicart
P
P
Institut Pasteur, SCME, Paris, France.
Testut
P
P
Schwartz
B
B
Llorens-Cortes
C
C
Perdomo
J J
JJ
Paulin
D
D
D000952:Antigens, Polyomavirus Transforming / Q000235:genetics*; D001483:Base Sequence; D015415:Biomarkers; D015815:Cell Adhesion Molecules / Q000378:metabolism*; D002461:Cell Line, Transformed; D002472:Cell Transformation, Viral / Q000235:genetics / Q000502:physiology*; D004730:Endothelium, Vascular / Q000166:cytology* / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D006801:Humans; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes / Q000235:genetics; D007703:Peptidyl-Dipeptidase A / Q000235:genetics / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011453:Prostaglandins / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D013917:Thrombin / Q000494:pharmacology; D014746:Vimentin / Q000235:genetics*
D000952:Antigens, Polyomavirus Transforming; D015415:Biomarkers; D015815:Cell Adhesion Molecules; D015345:Oligonucleotide Probes; D011453:Prostaglandins; D012333:RNA, Messenger; D014746:Vimentin; D007703:Peptidyl-Dipeptidase A; D013917:Thrombin; D013755:Tetradecanoylphorbol Acetate