Responses of human intestinal microvascular endothelial cells to shiga toxins 1 and 2 and pathogenesis of hemorrhagic colitis

Endothelial damage is characteristic of infection with Shiga toxin (Stx)-pr oducing Escherichia coli (STEC). Because Stx-mediated endothelial cell dama ge at the site of infection may lead to the characteristic hemorrhagic coli tis of STEC infection, we compared the effects of Stx1 and Stx2 on primary and transformed human intestinal microvascular endothelial cells (HIMEC) to those can macrovascular endothelial cells from human saphenous vein (HSVEC ). Adhesion molecule, interleukin-8 (IL-8), and Stx receptor expression, th e effects of cytokine activation and Stx toxins on these responses, and Stx 1 and Stx2 binding kinetics and bioactivity were measured. Adhesion molecul e and IL-8 expression increased in activated HIMEC, but these responses wer e blunted in the presence of toxin, especially in the presence of Stx1. In contrast to HSVEC, unstimulated HIMEC constitutively expressed Stx receptor at high levels, bound large amounts of toxin, were highly sensitive to tox in, and were not further sensitized by cytokines. Although the binding capa cities of HIMEC for Stx1 and Stx2 were comparable, the binding affinity of Stx1 to HIMEC was 50-fold greater than that of Stx2. Nonetheless, Stx2 was more toxic to HIMEC than an equivalent amount of Stx1. The decreased bindin g affinity and increased toxicity for HIMEC of Stx2 compared to those of St x1 may be relevant to the preponderance of Stx2-producing STEC involved in the pathogenesis of hemorrhagic colitis and its systemic complications. The differences between primary and transformed HIMEC in these responses were negligible. We conclude that transformed HIMEC lines could represent a simp le physiologically relevant model to study the role of Stx in the pathogene sis of hemorrhagic colitis.