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3. Interplay between enterohaemorrhagic Escherichia coli and nitric oxide during the infectious process

Author

Naïli, Ilham, Gardette, Marion, Garrivier, Annie, Daniel, Julien, Desvaux, Mickaël, Pizza, Mariagrazia, Gobert, Alain, Marchal, Thierry, Loukiadis, Estelle, Jubelin, Grégory, Microbiologie Environnement Digestif Santé (MEDIS), Université Clermont Auvergne [2017-2020] (UCA [2017-2020])-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), GlaxoSmithKline [Siena, Italy] (GSK), Laboratoire d'Ecologie Microbienne - UMR 5557 (LEM), Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Ecole Nationale Vétérinaire de Lyon (ENVL)-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Novartis Vaccines and Diagnostics [Siena], Interactions Cellules Environnement - UR (ICE), VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS), and European Union (EU) FP7-PEOPLE-607611INRAE institute VetAgro Sup institute DISCo program

Subjects
mouse models of infection, Virulence, Virulence Factors, Article, Bacterial Adhesion, Shiga Toxin, gut pathogen, Mice, Inbred C57BL, Mice, NG-Nitroarginine Methyl Ester, [SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology, nitric oxide, Enterohaemorrhagic E. coli, Enterohemorrhagic Escherichia coli, Host-Pathogen Interactions, Shiga toxins, Animals, Female, Renal Insufficiency, Enzyme Inhibitors, Escherichia coli Infections
Abstract

Enterohaemorrhagic Escherichia coli (EHEC) are bacterial pathogens responsible for life-threatening diseases in humans such as bloody diarrhoea and the hemolytic and uremic syndrome. To date, no specific therapy is available and treatments remain essentially symptomatic. In recent years, we demonstrated in vitro that nitric oxide (NO), a major mediator of the intestinal immune response, strongly represses the synthesis of the two cardinal virulence factors in EHEC, namely Shiga toxins (Stx) and the type III secretion system, suggesting NO has a great potential to protect against EHEC infection. In this study, we investigated the interplay between NO and EHEC in vivo using mouse models of infection. Using a NO-sensing reporter strain, we determined that EHEC sense NO in the gut of infected mice. Treatment of infected mice with a specific NOS inhibitor increased EHEC adhesion to the colonic mucosa but unexpectedly decreased Stx activity in the gastrointestinal tract, protecting mice from renal failure. Taken together, our data indicate that NO can have both beneficial and detrimental consequences on the outcome of an EHEC infection, and underline the importance of in vivo studies to increase our knowledge in host–pathogen interactions.

Published
2020
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4. Pushing the limits of STEC molecular risk assessment: identification of genes specifically induced in vivo as potential new genetic biomarkers

Author

Gardette, Marion, Le Hello, Simon, Garrivier, Annie, Rousseau, Valérie, Loukiadis, Estelle, Jubelin, Grégory, Microbiologie Environnement Digestif Santé (MEDIS), INRA Clermont-Ferrand-Theix-Université Clermont Auvergne [2017-2020] (UCA [2017-2020]), Institut Pasteur de Lille, Réseau International des Instituts Pasteur (RIIP), Laboratoire d'Ecologie Microbienne - UMR 5557 (LEM), Institut National de la Recherche Agronomique (INRA)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Ecole Nationale Vétérinaire de Lyon (ENVL)-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS), ProdInra, Migration, Institut National de la Recherche Agronomique (INRA)-Université Clermont Auvergne [2017-2020] (UCA [2017-2020]), Laboratoire d'Immunologie et Immunothérapie des Cancers (LIIC), École Pratique des Hautes Études (EPHE), and Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Université de Bourgogne (UB)

Subjects
[SDV] Life Sciences [q-bio], fluids and secretions, [SDV]Life Sciences [q-bio]
Abstract

National audience; Epidemiologic studies reveal that only few virulence factors are frequently associated with Shiga toxin-producing Escherichia coli (STEC) strains that cause severe disease in humans. Among them, Shiga toxins (Stx) is required for the development of the most severe symptoms in STEC infected patients but is not strictly sufficient to make an E. coli strain pathogenic to human. Indeed, many STEC strains isolated from rearing animals have never been implicated in human disease. It therefore appears crucial to increase our knowledge on STEC pathogenesis at the molecular level. We developed a RIVET (Recombination-based in vivo expression technology) strategy in order to identify STEC genes specifically induced in vivo during mouse infection. Construction and screening of a RIVET promoter library from the typical STEC O157:H7 strain EDL933 resulted in the identification of 31 in vivo induced (ivi) genes. Most but not all, have an attributed function and are involved in either metabolism or stress response pathways, indicating that STEC has to adapt to the intestinal ecosystem. Additionally, some identified ivi genes belong to the dispensable genome of E. coli and analysis of their distribution among E. coli strains revealed a strong prevalence for some of them specifically in STEC strains. We also observed correlations between ivi genes and virulence genes, serotypes and/or seropathotypes. Finally, we assessed their contribution to STEC pathogenesis by infecting mice with mutants inactivated for selected ivi genes. This work may thus help to improve STEC molecular risk assessment schemes by giving new insights into molecular aspects of STEC pathogenesis.

Published
2017

5. Role of nitric oxide in the outcome of EHEC infections

Author

Naili, Ilham, Gardette, Marion, Garrivier, Annie, Daniel, Julien, Rousseau, Valérie, Jubelin, Grégory, Microbiologie Environnement Digestif Santé (MEDIS), Institut National de la Recherche Agronomique (INRA)-Université Clermont Auvergne [2017-2020] (UCA [2017-2020]), ProdInra, Migration, and INRA Clermont-Ferrand-Theix-Université Clermont Auvergne [2017-2020] (UCA [2017-2020])

Subjects
[SDV] Life Sciences [q-bio], [SDV]Life Sciences [q-bio], ComputingMilieux_MISCELLANEOUS
Abstract

National audience

Published
2017

7. How to avoid the misinterpretation of Comet assay in Gammarus fossarum sperm as a biomarker of genotoxic impact: intrinsic variability and reference level

Author

Lacaze, Emilie, Gardette, Marion, Jubeaux, G., Devaux, Alain, Geffard, Olivier, Milieux aquatiques, écologie et pollutions (UR MALY), Centre national du machinisme agricole, du génie rural, des eaux et forêts (CEMAGREF), Institut National de la Recherche Agronomique (INRA), and Irstea Publications, Migration

Subjects
[SDE] Environmental Sciences, [SDE]Environmental Sciences, ComputingMilieux_MISCELLANEOUS
Abstract

International audience

Published
2010

8. Role of the Nitric Oxide Reductase NorVW in the Survival and Virulence of Enterohaemorrhagic Escherichia coli during Infection.

Author

Gardette, Marion, Daniel, Julien, Loukiadis, Estelle, and Jubelin, Grégory

Subjects
ESCHERICHIA coli diseases, REDUCTASES, NITRIC oxide, HEMOLYTIC-uremic syndrome, NITRIC-oxide synthases, ESCHERICHIA coli
Abstract

Enterohaemorrhagic Escherichia coli (EHEC) are bacterial pathogens responsible for life-threatening diseases in humans, such as hemolytic and uremic syndrome. It has been previously demonstrated that the interplay between EHEC and nitric oxide (NO), a mediator of the host immune innate response, is critical for infection outcome, since NO affects both Shiga toxin (Stx) production and adhesion to enterocytes. In this study, we investigated the role of the NO reductase NorVW in the virulence and fitness of two EHEC strains in a murine model of infection. We determined that the deletion of norVW in the strain O91:H21 B2F1 has no impact on its virulence, whereas it reduces the ability of the strain O157:H7 620 to persist in the mouse gut and to produce Stx. We also revealed that the fitness defect of strain 620 ΔnorVW is strongly attenuated when mice are treated with an NO synthase inhibitor. Altogether, these results demonstrate that the NO reductase NorVW participates in EHEC resistance against NO produced by the host and promotes virulence through the modulation of Stx synthesis. The contribution of NorVW in the EHEC infectious process is, however, strain-dependent and suggests that the EHEC response to nitrosative stress is complex and multifactorial. [ABSTRACT FROM AUTHOR]

Published
2020
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9. Interplay between enterohaemorrhagic Escherichia coli and nitric oxide during the infectious process.

Author

Naïli I, Gardette M, Garrivier A, Daniel J, Desvaux M, Pizza M, Gobert A, Marchal T, Loukiadis E, and Jubelin G

Subjects
Animals, Bacterial Adhesion drug effects, Enterohemorrhagic Escherichia coli pathogenicity, Enzyme Inhibitors administration & dosage, Female, Mice, Mice, Inbred C57BL, NG-Nitroarginine Methyl Ester administration & dosage, Nitric Oxide antagonists & inhibitors, Renal Insufficiency prevention & control, Shiga Toxin antagonists & inhibitors, Shiga Toxin metabolism, Virulence, Virulence Factors antagonists & inhibitors, Virulence Factors metabolism, Enterohemorrhagic Escherichia coli metabolism, Escherichia coli Infections metabolism, Host-Pathogen Interactions drug effects, Nitric Oxide metabolism
Abstract

Enterohaemorrhagic Escherichia coli (EHEC) are bacterial pathogens responsible for life-threatening diseases in humans such as bloody diarrhoea and the hemolytic and uremic syndrome. To date, no specific therapy is available and treatments remain essentially symptomatic. In recent years, we demonstrated in vitro that nitric oxide (NO), a major mediator of the intestinal immune response, strongly represses the synthesis of the two cardinal virulence factors in EHEC, namely Shiga toxins (Stx) and the type III secretion system, suggesting NO has a great potential to protect against EHEC infection. In this study, we investigated the interplay between NO and EHEC in vivo using mouse models of infection. Using a NO-sensing reporter strain, we determined that EHEC sense NO in the gut of infected mice. Treatment of infected mice with a specific NOS inhibitor increased EHEC adhesion to the colonic mucosa but unexpectedly decreased Stx activity in the gastrointestinal tract, protecting mice from renal failure. Taken together, our data indicate that NO can have both beneficial and detrimental consequences on the outcome of an EHEC infection, and underline the importance of in vivo studies to increase our knowledge in host-pathogen interactions.

Published
2020
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