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Shiga Toxin 2 and Lipopolysaccharide Induce Human Microvascular Endothelial Cells To Release Chemokines and Factors That Stimulate Platelet Function

Author:
Guessous, Fadila, Marcinkiewicz, Marek, Polanowska-Grabowska, Renata, Kongkhum, Sudawadee, Heatherly, Daniel, Obrig, Tom, Gear, Adrian R. L.
Source:
Infection and immunity 2005 v.73 no.12 pp. 8306-8316
ISSN:
0019-9567
Subject:
Shiga-like toxin 2, Shigella dysenteriae, adenosine diphosphate, agonists, chemokines, endothelial cells, enterohemorrhagic Escherichia coli, hemolytic uremic syndrome, humans, immunoassays, indomethacin, interleukin-8, lipopolysaccharides, platelet aggregation, prostaglandin synthase, secretion, thrombosis
Abstract:
Shiga toxins (Stxs) produced by Shigella dysenteriae type 1 and enterohemorrhagic Escherichia coli are the most common cause of hemolytic-uremic syndrome (HUS). It is well established that vascular endothelial cells, mainly those located in the renal microvasculature, are targets for Stxs. The aim of the present research was to evaluate whether E. coli-derived Shiga toxin 2 (Stx2) incubated with human microvascular endothelial cells (HMEC-1) induces release of chemokines and other factors that might stimulate platelet function. HMEC-1 were exposed for 24 h in vitro to Stx2, lipopolysaccharide (LPS), or the Stx2-LPS combination, and chemokine production was assessed by immunoassay. More interleukin-8 was released than stromal cell-derived factor 1[alpha] (SDF-1[alpha]) or SDF-1{szligbeta} and RANTES. The Stx2-LPS combination potentiated chemokine release, but Stx2 alone caused more release of SDF-1[alpha] at 24 h than LPS or Stx2-LPS did. In the presence of low ADP levels, HMEC-1 supernatants activated platelet function assessed by classical aggregometry, single-particle counting, granule secretion, P-selectin exposure, and the formation of platelet-monocyte aggregates. Supernatants from HMEC-1 exposed only to Stx2 exhibited enhanced exposure of platelet P-selectin and platelet-THP-1 cell interactions. Blockade of platelet cyclooxygenase by indomethacin prevented functional activation. The chemokine RANTES enhanced platelet aggregation induced by SDF-1[alpha], macrophage-derived chemokine, or thymus and activation-regulated chemokine in the presence of very low ADP levels. These data support the hypothesis that microvascular endothelial cells exposed to E. coli O157:H7-derived Stx2 and LPS release chemokines and other factors, which when combined with low levels of primary agonists, such as ADP, cause platelet activation and promote the renal thrombosis associated with HUS.
Agid:
267110