Differential cytotoxic actions of Shiga toxin 1 and Shiga toxin 2 on microvascular and macrovascular endothelial cells

Andreas Bauwens; Martina Bielaszewska; Björn Kemper; Patrik Langehanenberg; Gert von Bally; Rudolf Reichelt; Dennis Mulac; Hans-Ulrich Humpf; Alexander W. Friedrich; Kwang S. Kim; Helge Karch; Johannes Müthing

doi:10.1160/TH10-02-0140

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2011; 105(03): 515-528
DOI: 10.1160/TH10-02-0140

Endothelium and Vascular Development

Schattauer GmbH

Differential cytotoxic actions of Shiga toxin 1 and Shiga toxin 2 on microvascular and macrovascular endothelial cells

Andreas Bauwens

¹Institute for Hygiene, University of Münster, Münster, Germany

,

Martina Bielaszewska

¹Institute for Hygiene, University of Münster, Münster, Germany

,

Björn Kemper

²Center for Biomedical Optics and Photonics, University of Münster, Münster, Germany

,

Patrik Langehanenberg

²Center for Biomedical Optics and Photonics, University of Münster, Münster, Germany

,

Gert von Bally

²Center for Biomedical Optics and Photonics, University of Münster, Münster, Germany

,

Rudolf Reichelt

³Institute for Medical Physics and Biophysics, University of Münster, Münster, Germany

,

Dennis Mulac

⁴Institute for Food Chemistry, University of Münster, Münster, Germany

,

Hans-Ulrich Humpf

⁴Institute for Food Chemistry, University of Münster, Münster, Germany

,

Alexander W. Friedrich

¹Institute for Hygiene, University of Münster, Münster, Germany

,

Kwang S. Kim

⁵Division of Pediatric Infectious Diseases, Johns Hopkins University School of Medicine, David M. Rubenstein Child Health Building, Baltimore, Maryland, USA

,

Helge Karch

¹Institute for Hygiene, University of Münster, Münster, Germany

,

Johannes Müthing

¹Institute for Hygiene, University of Münster, Münster, Germany

› Author Affiliations

Abstract

Summary

Shiga toxin (Stx)-mediated injury to vascular endothelial cells in the kidneys, brain and other organs underlies the pathogenesis of haemolytic uraemic syndrome (HUS) caused by enterohaemorrhagic Escherichia coli (EHEC). We present a direct and comprehensive comparison of cellular injury induced by the two major Stx types, Stx1 and Stx2, in human brain microvascular endothelial cells (HBMECs) and EA.hy 926 macro-vascular endothelial cells. Scanning electron microscopy of microcarrier-based cell cultures, digital holographic microscopy of living single cells, and quantitative apoptosis/necrosis assays demonstrate that Stx1 causes both necrosis and apoptosis, whereas Stx2 induces almost exclusively apoptosis in both cell lines. Moreover, microvascular and macrovascular endothelial cells have different susceptibilities to the toxins: EA.hy 926 cells are slightly, but significantly (~ 10 times) more susceptible to Stx1, whereas HBMECs are strikingly (≥ 1,000 times) more susceptible to Stx2. These findings have implications in the pathogenesis of HUS, and suggest the existence of yet to be delineated Stx type-specific mechanisms of endothelial cell injury beyond inhibition of protein bio-synthesis.

Keywords

Shiga toxin - endothelial cells - enterohaemorrhagic E. coli - single cell analysis - apoptosis - necrosis

Full Text

References

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