Immortalized human endothelial cells have a decreased response to IL-1 secondary to an IL-1 receptor defective expression restored by corticosteroids.
A human endothelial cell line is a convenient tool for exploring cell physiology and testing drugs and toxics. Several attempts have been made using SV40 to immortalize endothelial cells. We used human umbilical vein endothelial cells (HUVEC) transformed with a construct made of promoter of the vimentin gene and SV40 Tag. The proliferation of immortalized vascular endothelial cells (IVEC), as measured by [methyl-3H]thymidine incorporation, was compared to that of HUVEC in the presence of endothelial cell growth factor and cytokines: tumor necrosis factor-alpha (TNF-alpha), interleukin-1beta (IL-1beta) and interferon-gamma (IFN-gamma). Inhibition of [methyl-3H]thymidine incorporation by IL-1beta was lower than that observed with HUVEC, while TNF-alpha reduced the proliferation of IVEC and HUVEC to similar extents. Induction of intercellular adhesion molecule (ICAM-1), vascular cell adhesion molecule (VCAM-1) and E-selectin by TNF-alpha, measured by a radiometric technique, was similar in IVEC and HUVEC, while the induction of E-selectin by IL-1beta on IVEC was limited and significantly different from that observed on HUVEC (p<0.001). The number of 125I-IL-1beta binding sites on IVEC is 3-fold less than on HUVEC and the IL-1beta receptor number was reduced. Dexamethasone treatment of IVEC restored their reactivity to IL-1beta and corrected the IL-1beta binding and the receptor number. These results showed that the introduction of SV40 gene not only immortalized the cell but also altered IL-1 receptor expression. This alteration may be improved by addition of corticosteroids to the cell culture, which extends the possibility of using IVEC as a model of endothelial cells.
Wautier
M P
MP
Laboratoire de Biologie Vasculaire et Cellulaire, UFR Lariboisière St Louis, Paris 7, INTS, France.
Chappey
O
O
Vicart
P
P
Paulin
D
D
Wautier
J L
JL
D002461:Cell Line, Transformed; D003907:Dexamethasone / Q000494:pharmacology*; D019040:E-Selectin / Q000096:biosynthesis; D004730:Endothelium, Vascular / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D005938:Glucocorticoids / Q000494:pharmacology*; D006131:Growth Inhibitors / Q000494:pharmacology; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000096:biosynthesis; D007371:Interferon-gamma / Q000494:pharmacology; D007375:Interleukin-1 / Q000494:pharmacology*; D017472:Receptors, Interleukin-1 / Q000096:biosynthesis*; D016028:Receptors, Leukocyte-Adhesion / Q000096:biosynthesis; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D014471:Umbilical Veins; D019010:Vascular Cell Adhesion Molecule-1 / Q000096:biosynthesis
D019040:E-Selectin; D005938:Glucocorticoids; D006131:Growth Inhibitors; D007375:Interleukin-1; D017472:Receptors, Interleukin-1; D016028:Receptors, Leukocyte-Adhesion; D014409:Tumor Necrosis Factor-alpha; D019010:Vascular Cell Adhesion Molecule-1; D018799:Intercellular Adhesion Molecule-1; D003907:Dexamethasone; D007371:Interferon-gamma