Escherichia coli verotoxin binding to human paediatric glomerular mesangial cells

Abstract

Haemolytic uraemic syndrome (HUS) remains the leading cause of acute renal failure in children. Although anEscherichia coli-produced verotoxin (VT) has been implicated in the pathogenesis of HUS, the precise mechanisms of disease are not well defined. We hypothesise that the pathogenesis of renal failure in HUS includes the binding ofE. coli VT to the glomerular mesangial cell, with consequent effects on renal function. Using human paediatric mesangial cells. we studied the binding and biological effects of the purified verotoxin VT-1. We isolated, purified and characterised paediatric glomerular mesangial cells. The mesangial cells were characterised by their immunoreactivity with both smooth muscle actin and vimentin antibodies, and lack of immunoreactivity with cytokeratin or factor VIII antibodies. Using an fluorescein isothiocyanate-conjugated VT (10⁻⁷–10⁻⁸ M), we demonstrated specific binding to the mesangial cell membrane by immunofluorescence microscopy. We also demonstrated a dose-dependent inhibition of mesangial cell mitogenesis at concentrations from 10⁻⁹ to 10⁻¹⁷ M. Our data demonstrate that VT-1 binds to paediatric human glomerular mesangial cells and this binding results in specific biological actions, including an inhibition of cell mitogenesis.