Tumor Necrosis Factor and Interleukin-1 Induce Expression of the Verocytotoxin Receptor Globotriaosylceramide on Human Endothelial Cells: Implications for the Pathogenesis of the Hemolytic Uremic Syndrome

Abstract

AbstractThe epidemic form of the hemolytic uremic syndrome (HUS), beginning with an acute gastroenteritis, has been associated with a verocytotoxin-producing Escherichia coli infection. The endothelial cell is believed to play an important role in the pathogenesis of HUS. Endothelial cell damage by verocy-totoxin-1 (VT-1) in vitro is potentiated by the additional exposure of inflammatory mediators, such as tumor necrosis factor-α (TNF-α). Preincubation of human umbilical vein endothelial cells (HUVEC) with TNF-α resulted in a 10- to 100-fold increase of specific binding sites for 125I-VT-1. Furthermore, interleukin-1 (IL-1), lymphotoxin (TNF-β), and lipopolysaccharide (LPS) also markedly increase VT-1 binding. Several hours’ exposure to TNF-α was enough to enhance the number of VT-1 receptors on the endothelial cells for 2 days. The TNF-α-induced increase in VT-1 binding could be inhibited by simultaneous addition of the protein synthesis inhibitor cycloheximide. Glycolipid extracts of TNF-α-treated cells tested on thin-layer chromatography demonstrated an increase of globotriaosylceramide (GbOse3cer), a functional receptor for VT-1, which suggests that preincubation of human endothelial cells with TNF-α leads to an increase in GbOse3cer synthesis in these cells. We conclude from this study that TNF-α and IL-1 induce one (or more) enzyme(s) that is (are) rate-limiting in the synthesis of the glycolipid VT-1 receptor, GbOse3cer. These in vitro studies suggest that, in addition to VT-1, inflammatory mediators play an important role in the pathogenesis of HUS.