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Virulence Factor Cargo and Host Cell Interactions of Shiga Toxin-Producing Escherichia coli Outer Membrane Vesicles

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Shiga Toxin-Producing E. coli

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2291))

Abstract

Outer membrane vesicles (OMVs), nanoparticles released by Shiga toxin-producing Escherichia coli (STEC), have been identified as novel efficient virulence tools of these pathogens. STEC O157 OMVs carry a cocktail of virulence factors including Shiga toxin 2a (Stx2a), cytolethal distending toxin V (CdtV), EHEC hemolysin, flagellin, and lipopolysaccharide. OMVs are taken up by human intestinal epithelial and microvascular endothelial cells, the major targets during STEC infection, and deliver the virulence factors into host cells. There the toxins separate from OMVs and are trafficked via different pathways to their target compartments, i.e., the cytosol (Stx2a-A subunit), nucleus (CdtV-B subunit), and mitochondria (EHEC hemolysin). This leads to a toxin-specific host cell injury and ultimately apoptotic cell death. Besides their cytotoxic effects, STEC OMVs trigger an inflammatory response via their lipopolysaccharide and flagellin components. In this chapter, we describe methods for the isolation and purification of STEC OMVs, for the detection of OMV-associated virulence factors, and for the analysis of OMV interactions with host cells including OMV cellular uptake and intracellular trafficking of OMVs and OMV-delivered toxins.

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Acknowledgments

The work reported in this chapter was supported by the German Research Foundation grant SFB1009, projects B04 (to A. M.) and B03 (to C. R.), and by grant from the Interdisciplinary Center of Clinical Research (IZKF) Münster De2/006/20 (to P. D.).

We are much indebted to Prof. em. Dr. Dr. h. c. Helge Karch for his long-lasting support of the research on STEC outer membrane vesicles.

We thank Roland Lloubes (CNRS-Aix-Marseille Université, Marseille, France), Xiaohua He (United States Department of Agriculture, Albany, CA, USA), Herbert Schmidt (University of Hohenheim, Stuttgart, Germany), Angelika Fruth (Robert Koch Institute, Wernigerode, Germany), Volker Gerke (University of Münster, Germany), and Kwang Sik Kim (Johns Hopkins University School of Medicine, Baltimore, MD, USA) for providing us with antibodies against OmpA, Stx2a, EHEC-Hly, LPS and flagellin, LAMP3/CD63, and the HBMEC line, respectively.

Author information

Authors and Affiliations

  1. National Reference Laboratory for E. coli and Shigellae, National Institute of Public Health, Prague, Czech Republic

    Martina Bielaszewska

  2. Institute for Hygiene, University of Münster, Münster, Germany

    Martina Bielaszewska, Andreas Bauwens & Alexander Mellmann

  3. Institute for Infectiology, Center for Molecular Biology of Inflammation (ZMBE), University of Münster, Münster, Germany

    Lilo Greune, Petra Dersch & Christian Rüter

Authors
  1. Martina Bielaszewska
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  2. Lilo Greune
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  3. Andreas Bauwens
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  4. Petra Dersch
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  5. Alexander Mellmann
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  6. Christian Rüter
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Corresponding author

Correspondence to Martina Bielaszewska .

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Editors and Affiliations

  1. University of East Anglia, Norwich Medical School, Norwich, UK

    Stephanie Schüller

  2. National Reference Laboratory for E. coli and Shigellae, National Institute of Public Health, Prague, Czech Republic

    Martina Bielaszewska

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Bielaszewska, M., Greune, L., Bauwens, A., Dersch, P., Mellmann, A., Rüter, C. (2021). Virulence Factor Cargo and Host Cell Interactions of Shiga Toxin-Producing Escherichia coli Outer Membrane Vesicles. In: Schüller, S., Bielaszewska, M. (eds) Shiga Toxin-Producing E. coli . Methods in Molecular Biology, vol 2291. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1339-9_8

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  • DOI: https://doi.org/10.1007/978-1-0716-1339-9_8

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1338-2

  • Online ISBN: 978-1-0716-1339-9

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