Tremoli and Yale Nemerson
نویسندگان
چکیده
This study was undertaken to characterize tissue factor (TF) induction, localization, and functional activity in cultured human umbilical vein endothelial cells (HUVECs) exposed to recombinant vascular endothelial growth factor (rVEGF) and recombinant tumor necrosis factor-a (rTNF-a). rVEGF (1 nmol/L) and rTNF-a (500 U/mL) synergistically increased TF mRNA, protein, and total activity, as measured in cell lysates. To examine surface TF expression, living cells were treated with antibody to TF and examined microscopically. Almost no staining was seen in control cells or cells treated with a single agent. In contrast, cells treated with both agonists showed intense membrane staining with surface patches, appearing as buds by confocal microscopy. To determine surface TF activity, studies were performed using a parallel-plate flow chamber, which allows detection of factor Xa generation on living cells. rVEGF and rTNF-a induced little surface TF activity (0.03260.008 and 0.01460.008 fmol/cm, respectively). In combination, they significantly increased TF expression on the cell surface (0.42960.094 fmol/cm, P,0.05). These data indicate that the synergistic effect of rVEGF and rTNF-a is necessary to generate functional TF on the surface of endothelial cells. The requirement for multiple agonists to expose active TF may serve to protect endothelial cells from acting as a procoagulant surface, even under conditions of cell perturbation. (Arterioscler Thromb Vasc Biol. 1999;19:531-537.)
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