Biosynthesis of IL-8 and endothelin-1,2 by immortal simian virus 40 tsA-transformed human endothelial cells

Immortal human umbilical cord vein endothelial cells (HUVEC-SV40) established in our laboratory by gene transfection of plasmid pSVts58neo were analyzed for their biological activities. The growth and cellular viability of these immortal cells were temperature dependent. Typical bioactive products of endothelial cells, IL-8 and endothelins, were released into the culture medium as in the case of mortal cells. The production of IL-8 was temperature dependent and significantly increased in cells incubated at 37 degrees C as compared with those incubated at 33 degrees C. Such a temperature dependence was not observed in endothelin production. Stimulation of cells with both TNF-alpha and IFN-gamma increased the biosynthesis of IL-8 or endothelins significantly (p<0.05). From these results, it became clear that immortal HUVEC-SV40 cells also secrete IL-8 or endothelins in the same manner as mortal cells, and that they react to the stimulation by cytokines to promote IL-8 or endothelin production.