Immortalized human endothelial cells have a decreased response to IL-1 secondary to an IL-1 receptor defective expression restored by corticosteroids |
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Authors: | M-P Wautier O Chappey P Vicart D Paulin J-L Wautier |
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Institution: | (1) Laboratoire de Biologie Vasculaire et Cellulaire, UFR Lariboisière, St Louis Paris, 7, INTS, France;(2) Faculté de Médecine Pitié Salpêtrière, CNRS URA, 2115 Paris, France;(3) Laboratoire de Biologie Moléculaire de la Différenciation, UFR Biochimie, Paris, 7, France |
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Abstract: | 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- (TNF- ), interleukin-1 (IL-1 ) and interferon- (IFN- ). Inhibition of methyl-3H]thymidine incorporation by IL-1 was lower than that observed with HUVEC, while TNF- 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- , measured by a radiometric technique, was similar in IVEC and HUVEC, while the induction of E-selectin by IL-1 on IVEC was limited and significantly different from that observed on HUVEC (p<0.001). The number of 125I-IL-1 binding sites on IVEC is 3-fold less than on HUVEC and the IL-1 receptor number was reduced. Dexamethasone treatment of IVEC restored their reactivity to IL-1 and corrected the IL-1 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. |
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Keywords: | endothelial cells immortalized vascular endothelial cells interleukin-1 receptor SV40 T antigen |
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