Immortalized human endothelial cells have a decreased response to IL-1 secondary to an IL-1 receptor defective expression restored by corticosteroids

被引:5
作者
Wautier, MP
Chappey, O
Vicart, P
Paulin, D
Wautier, JL
机构
[1] Inst Natl Transfus Sanguine, UFR Lariboisiere St Louis Paris 7, Lab Biol Vasc & Cellulaire, F-75739 Paris 15, France
[2] Fac Med Pitie Salpetriere, CNRS, URA 2115, Paris, France
[3] UFR Biochim Paris 7, Lab Biol Mol Differenciat, Paris, France
关键词
endothelial cells; immortalized vascular endothelial cells; interleukin-1; receptor; SV40 T antigen;
D O I
10.1023/A:1007629403123
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
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-H-3]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-1 beta (IL-1 beta) and interferon-gamma (IFN-gamma). Inhibition of [methyl-H-3]thymidine incorporation by IL-1 beta 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-1 beta on IVEC was limited and significantly different from that observed on HUVEC (p < 0.001). The number of I-125-IL-1 beta binding sites on IVEC is 3-fold less than on HUVEC and the IL-1 beta receptor number was reduced. Dexamethasone treatment of IVEC restored their reactivity to IL-1 beta and corrected the IL-1 beta 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.
引用
收藏
页码:153 / 161
页数:9
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