Presence of DNA in OMVs (intact, either DNase-untreated or DNase-treated, or lysed after the DNase treatment 14 ) was analysed with Quant-iT PicoGreen dsDNA Assay (Molecular Probes) 14 . PCRs for virulence genes detection (listed in Supplementary Table S2) were performed with published primers 2,52,53 in 25 ? l volume containing 2.5 ? l of OMVs (either non-fractionated, DNase-untreated or DNase-treated 14 , or pools of OptiPrep fractions References 1. Karch, H. et al. The enemy within us: lessons from the 2011 European Escherichia coli O104:H4 outbreak, EMBO Mol. Med, vol.4, pp.841-848, 2012. ,
Characterisation of the Escherichia coli strain associated with an outbreak of haemolytic uraemic syndrome in Germany, 2011: a microbiological study, The Lancet Infectious Diseases, vol.11, issue.9, pp.671-676, 2011. ,
DOI : 10.1016/S1473-3099(11)70165-7
Prospective Genomic Characterization of the German Enterohemorrhagic Escherichia coli O104:H4 Outbreak by Rapid Next Generation Sequencing Technology, PLoS ONE, vol.61, issue.7, p.22751, 2011. ,
DOI : 10.1371/journal.pone.0022751.s002
Strain Causing an Outbreak of Hemolytic???Uremic Syndrome in Germany, New England Journal of Medicine, vol.365, issue.8, pp.709-717, 2011. ,
DOI : 10.1056/NEJMoa1106920
Virulence of the Shiga Toxin Type 2-Expressing Escherichia coli O104:H4 German Outbreak Isolate in Two Animal Models, Infection and Immunity, vol.81, issue.5, pp.1562-1574, 2013. ,
DOI : 10.1128/IAI.01310-12
Lability of the pAA Virulence Plasmid in Escherichia coli O104:H4: Implications for Virulence in Humans, PLoS ONE, vol.17, issue.6, p.66717, 2013. ,
DOI : 10.1371/journal.pone.0066717.s001
Autotransporters but not pAA are critical for rabbit colonization by Shiga toxin-producing Escherichia coli O104:H4, Nature Communications, vol.59, p.3080, 2014. ,
DOI : 10.1016/j.chom.2011.07.007
The Presence of the pAA Plasmid in the German O104:H4 Shiga Toxin Type 2a (Stx2a)-Producing Enteroaggregative Escherichia coli Strain Promotes the Translocation of Stx2a Across an Epithelial Cell Monolayer, Journal of Infectious Diseases, vol.210, issue.12, pp.1909-1919, 2014. ,
DOI : 10.1093/infdis/jiu399
Protein secretion systems and adhesins: The molecular armory of Gram-negative pathogens, International Journal of Medical Microbiology, vol.297, issue.6, pp.401-415, 2007. ,
DOI : 10.1016/j.ijmm.2007.03.017
Outer membrane vesicles function as offensive weapons in host???parasite interactions, Microbes and Infection, vol.12, issue.11, pp.791-798, 2010. ,
DOI : 10.1016/j.micinf.2010.05.008
Virulence and Immunomodulatory Roles of Bacterial Outer Membrane Vesicles, Microbiology and Molecular Biology Reviews, vol.74, issue.1, pp.81-94, 2010. ,
DOI : 10.1128/MMBR.00031-09
Long-Distance Delivery of Bacterial Virulence Factors by Pseudomonas aeruginosa Outer Membrane Vesicles, PLoS Pathogens, vol.280, issue.4, p.1000382, 2009. ,
DOI : 10.1371/journal.ppat.1000382.s004
O157:H7, FEMS Immunology & Medical Microbiology, vol.63, issue.3, pp.427-434, 2011. ,
DOI : 10.1111/j.1574-695X.2011.00869.x
Enterohemorrhagic Escherichia coli Hemolysin Employs Outer Membrane Vesicles to Target Mitochondria and Cause Endothelial and Epithelial Apoptosis, PLoS Pathogens, vol.9, issue.1, p.1003797, 2013. ,
DOI : 10.1371/journal.ppat.1003797.s009
Dynasore, a Cell-Permeable Inhibitor of Dynamin, Developmental Cell, vol.10, issue.6, pp.839-850, 2006. ,
DOI : 10.1016/j.devcel.2006.04.002
Mis-assembly of clathrin lattices on endosomes reveals a regulatory switch for coated pit formation, The Journal of Cell Biology, vol.123, issue.5, pp.1107-1117, 1993. ,
DOI : 10.1083/jcb.123.5.1107
Hypertonic media inhibit receptor-mediated endocytosis by blocking clathrin-coated pit formation, The Journal of Cell Biology, vol.108, issue.2, pp.389-400, 1989. ,
DOI : 10.1083/jcb.108.2.389
Filipin-dependent Inhibition of Cholera Toxin: Evidence for Toxin Internalization and Activation through Caveolae-like Domains, The Journal of Cell Biology, vol.26, issue.4, pp.905-915, 1998. ,
DOI : 10.1083/jcb.123.5.1107
Cholesterol controls the clustering of the glycophospholipid-anchored membrane receptor for 5-methyltetrahydrofolate, The Journal of Cell Biology, vol.111, issue.6, pp.2931-2938, 1990. ,
DOI : 10.1083/jcb.111.6.2931
Transducible TAT-HA fusogenic peptide enhances escape of TAT-fusion proteins after lipid raft macropinocytosis, Nature Medicine, vol.10, issue.3, pp.310-315, 2004. ,
DOI : 10.1038/nm996
Vero response to a cytotoxin of Escherichia coli, Infect. Immun, vol.18, pp.775-779, 1977. ,
Maturational regulation of globotriaosylceramide, the Shiga-like toxin 1 receptor, in cultured human gut epithelial cells., Journal of Clinical Investigation, vol.96, issue.3, pp.1328-1335, 1995. ,
DOI : 10.1172/JCI118168
Interaction of Shiga toxin from Escherichia coli with human intestinal epithelial cell lines and explants: Stx2 induces epithelial damage in organ culture, Cellular Microbiology, vol.64, issue.3, pp.289-301, 2004. ,
DOI : 10.1016/S0378-4274(97)03879-4
A Polyclonal Antibody Based Immunoassay Detects Seven Subtypes of Shiga Toxin 2 Produced by Escherichia coli in Human and Environmental Samples, PLoS ONE, vol.336, issue.10, p.76368, 2013. ,
DOI : 10.1371/journal.pone.0076368.s001
Enteroaggregative Escherichia coli flagellin-induced interleukin-8 secretion requires Toll-like receptor 5-dependent p38 MAP kinase activation, Immunology, vol.168, issue.4, pp.651-660, 2004. ,
DOI : 10.1128/IAI.69.7.4424-4429.2001
Recognition of lipopolysaccharide pattern by TLR4 complexes, Experimental & Molecular Medicine, vol.169, issue.12, p.66, 2013. ,
DOI : 10.1038/nsmb.2106
Binding of polymyxin B to the lipid A portion of bacterial lipopolysaccharides, Immunochemistry, vol.13, issue.10, pp.813-818, 1976. ,
DOI : 10.1016/0019-2791(76)90181-6
Naturally Produced Outer Membrane Vesicles from Pseudomonas aeruginosa Elicit a Potent Innate Immune Response via Combined Sensing of Both Lipopolysaccharide and Protein Components, Infection and Immunity, vol.78, issue.9, pp.3822-3831, 2010. ,
DOI : 10.1128/IAI.00433-10
Export of virulence genes and Shiga toxin by membrane vesicles of Escherichia coli O157:H7, App. Environ. Microbiol, vol.65, pp.1843-1848, 1999. ,
Phage regulatory circuits and virulence gene expression, Current Opinion in Microbiology, vol.8, issue.4, pp.459-465, 2005. ,
DOI : 10.1016/j.mib.2005.06.001
Globotriaosyl ceramide receptor function - Where membrane structure and pathology intersect, FEBS Letters, vol.246, issue.9, pp.1879-1886, 2010. ,
DOI : 10.1016/j.febslet.2009.11.089
Role of lipid rafts in Shiga toxin 1 interaction with the apical surface of Caco-2 cells, J. Cell. Sci, vol.114, pp.4025-4031, 2001. ,
The glycosphingolipid globotriaosylceramide in the metastatic transformation of colon cancer, Proc. Natl. Acad. Sci. USA. 102, pp.19087-19092, 2005. ,
DOI : 10.1073/pnas.0506474102
Enterotoxigenic Escherichia coli vesicles target toxin delivery into mammalian cells, The EMBO Journal, vol.115, issue.23, pp.4538-4549, 2004. ,
DOI : 10.1083/jcb.141.4.917
outer membrane vesicles which are internalized by human intestinal epithelial cells, FEBS Letters, vol.5, issue.9, pp.1357-1362, 2011. ,
DOI : 10.1016/j.febslet.2011.04.017
Role of Shiga toxin versus H7 flagellin in enterohaemorrhagic Escherichia coli signalling of human colon epithelium in vivo, Cellular Microbiology, vol.6, issue.5, pp.869-879, 2006. ,
DOI : 10.1128/IAI.71.4.2120-2129.2003
Macropinocytosis in Shiga toxin 1 uptake by human intestinal epithelial cells and transcellular transcytosis, AJP: Gastrointestinal and Liver Physiology, vol.296, issue.1, pp.78-92, 2009. ,
DOI : 10.1152/ajpgi.90347.2008
???O157:H7 and O104:H4, Cellular Microbiology, vol.78, issue.8, pp.1255-1266, 2014. ,
DOI : 10.1111/cmi.12281
Shiga Toxin Translocation across Intestinal Epithelial Cells Is Enhanced by Neutrophil Transmigration, Infection and Immunity, vol.69, issue.10, pp.6148-6155, 2001. ,
DOI : 10.1128/IAI.69.10.6148-6155.2001
A Novel Mechanism of Bacterial Toxin Transfer within Host Blood Cell-Derived Microvesicles, PLOS Pathogens, vol.12, issue.2, p.1004619, 2015. ,
DOI : 10.1371/journal.ppat.1004619.s004
The Induction of Apoptosis by Shiga Toxins and Ricin, Curr. Top. Microbiol. Immunol, vol.357, pp.137-178, 2012. ,
DOI : 10.1007/82_2011_155
Apoptosis of renal cortical cells in the hemolytic-uremic syndrome: in vivo and in vitro studies, Infect. Immun, vol.66, pp.636-644, 1998. ,
Direct acute tubular damage contributes to Shigatoxin-mediated kidney failure, The Journal of Pathology, vol.31, issue.1, pp.120-133, 2014. ,
DOI : 10.1002/path.4388
Role of EHEC O157:H7 virulence factors in the activation of intestinal epithelial cell NF-??B and MAP kinase pathways and the upregulated expression of interleukin 8, Cellular Microbiology, vol.6, issue.10, pp.635-648, 2002. ,
DOI : 10.1046/j.1462-5822.2002.00218.x
Enhanced CXC Chemokine Responses of Human Colonic Epithelial Cells to Locus of Enterocyte Effacement-Negative Shiga-Toxigenic Escherichia coli, Infection and Immunity, vol.71, issue.10, pp.5623-5632, 2003. ,
DOI : 10.1128/IAI.71.10.5623-5632.2003
Shiga Toxin 2 and Flagellin from Shiga-Toxigenic Escherichia coli Superinduce Interleukin-8 through Synergistic Effects on Host Stress-Activated Protein Kinase Activation, Infection and Immunity, vol.78, issue.7, pp.2984-2994, 2010. ,
DOI : 10.1128/IAI.00383-10
Enteroaggregative Escherichia coli expresses a novel flagellin that causes IL-8 release from intestinal epithelial cells, Journal of Clinical Investigation, vol.105, issue.12, pp.1769-1777, 2000. ,
DOI : 10.1172/JCI8892
Shiga toxins stimulate secretion of interleukin-8 from intestinal epithelial cells, Infect. Immun, vol.67, pp.5985-5993, 1999. ,
Two toxin-converting phages from Escherichia coli O157:H7 strain 933 encode antigenically distinct toxins with similar biologic activities, Infect. Immun, vol.53, pp.135-140, 1986. ,
Serotyping of Escherichia coli in Methods in Microbiology, pp.43-112, 1984. ,
Pal Lipoprotein of Escherichia coli Plays a Major Role in Outer Membrane Integrity, Journal of Bacteriology, vol.184, issue.3, pp.754-759, 2002. ,
DOI : 10.1128/JB.184.3.754-759.2002
Short report: high prevalence of serine protease autotransporter cytotoxins among strains of enteroaggregative Escherichia coli, Am. J. Trop. Med. Hyg, vol.80, pp.294-301, 2009. ,
Genomic Variability of O Islands Encoding Tellurite Resistance in Enterohemorrhagic Escherichia coli O157:H7 Isolates, Journal of Bacteriology, vol.184, issue.17, pp.4690-4698, 2002. ,
DOI : 10.1128/JB.184.17.4690-4698.2002
Differential cytotoxic actions of Shiga toxin 1 and Shiga toxin 2 on microvascular and macrovascular endothelial cells, Thrombosis and Haemostasis, vol.105, issue.3, pp.515-528, 2011. ,
DOI : 10.1160/TH10-02-0140
A rapid and simple method for measuring thymocyte apoptosis by propidium iodide staining and flow cytometry, Journal of Immunological Methods, vol.139, issue.2, pp.271-279, 1991. ,
DOI : 10.1016/0022-1759(91)90198-O
Cytolethal Distending Toxin from Shiga Toxin-Producing Escherichia coli O157 Causes Irreversible G2/M Arrest, Inhibition of Proliferation, and Death of Human Endothelial Cells, Infection and Immunity, vol.73, issue.1, pp.552-562, 2005. ,
DOI : 10.1128/IAI.73.1.552-562.2005
Quantitative Stain-Free and Continuous Multimodal Monitoring of Wound Healing In Vitro with Digital Holographic Microscopy, PLoS ONE, vol.7, issue.9, p.7317, 2014. ,
DOI : 10.1371/journal.pone.0107317.s004
Autofocussing in digital holographic microscopy, 3D Res, vol.2, p.1004, 2011. ,