Differential cytotoxic actions of Shiga toxin 1 and Shiga toxin 2 on microvascular and macrovascular endothelial cells
Andreas Bauwens
1
Institute for Hygiene, University of Münster, Münster, Germany
,
Martina Bielaszewska
1
Institute for Hygiene, University of Münster, Münster, Germany
,
Björn Kemper
2
Center for Biomedical Optics and Photonics, University of Münster, Münster, Germany
,
Patrik Langehanenberg
2
Center for Biomedical Optics and Photonics, University of Münster, Münster, Germany
,
Gert von Bally
2
Center for Biomedical Optics and Photonics, University of Münster, Münster, Germany
,
Rudolf Reichelt
3
Institute for Medical Physics and Biophysics, University of Münster, Münster, Germany
,
Dennis Mulac
4
Institute for Food Chemistry, University of Münster, Münster, Germany
,
Hans-Ulrich Humpf
4
Institute for Food Chemistry, University of Münster, Münster, Germany
,
Alexander W. Friedrich
1
Institute for Hygiene, University of Münster, Münster, Germany
,
Kwang S. Kim
5
Division of Pediatric Infectious Diseases, Johns Hopkins University School of Medicine, David M. Rubenstein Child Health Building, Baltimore, Maryland, USA
,
Helge Karch
1
Institute for Hygiene, University of Münster, Münster, Germany
,
Johannes Müthing
1
Institute for Hygiene, University of Münster, Münster, Germany
› Author AffiliationsFinancial support: This work was supported by grants from the Deutsche Forschungsgemeinschaft (DFG) MU 845/4–1, FR 2569/1–1, the Graduate School “Molecular Interactions of Pathogens with Biotic and Abiotic Surfaces“ (GRK 1409), and the program “Infections of the Endothelium“ SPP 1130 project KA 717/4–3, by a grant from the Interdisciplinary Center of Clinical Research (IZKF) Münster, project no. Ka2/061/04, and by a grant from the German Federal Ministry of Education and Research (BMBF), project FKZ 13N8183 within the research focus program “Biophotonics“.
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
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