Expression of Tumor Necrosis Factor-α in Cultured Human Endothelial Cells Stimulated With Lipopolysaccharide or Interleukin-1α

Abstract
—Tumor-necrosis factor-α (TNF-α) is a proinflammatory cytokine with a wide variety of biological effects. The most important source of this cytokine is monocytes/macrophages. It is a potent agonist in the activation of endothelial cells; however, the precise role of endothelial cells as a source of TNF-α is not known. In the present study, we addressed the possibility that TNF-α is produced by cultured human umbilical vein endothelial cells (HUVEC) stimulated with factors such as lipopolysaccharide (LPS) or interleukin-1α (IL-1α). LPS and IL-1α induced expression of TNF-α mRNA in HUVEC. IL-1α induced expression and secretion of TNF-α protein, but LPS did not induce production of TNF-α protein. Most of the TNF-α protein in cell lysate was found in the membrane fraction. The mRNA for TNF-α–converting enzyme (TACE) was expressed in unstimulated HUVEC, and its level was not altered by treatment with LPS or IL-1α. Transfection of HUVEC with full-length cDNA encoding the precursor TNF-α enhanced secreti...