Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Shigella-mediated oxygen depletion is essential for intestinal mucosa colonization

Abstract

Pathogenic enterobacteria face various oxygen (O2) levels during intestinal colonization from the O2-deprived lumen to oxygenated tissues. Using Shigella flexneri as a model, we have previously demonstrated that epithelium invasion is promoted by O2 in a type III secretion system-dependent manner. However, subsequent pathogen adaptation to tissue oxygenation modulation remained unknown. Assessing single-cell distribution, together with tissue oxygenation, we demonstrate here that the colonic mucosa O2 is actively depleted by S. flexneri aerobic respiration—and not host neutrophils—during infection, leading to the formation of hypoxic foci of infection. This process is promoted by type III secretion system inactivation in infected tissues, favouring colonizers over explorers. We identify the molecular mechanisms supporting infectious hypoxia induction, and demonstrate here how enteropathogens optimize their colonization capacity in relation to their ability to manipulate tissue oxygenation during infection.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

Fig. 1: Hypoxia is specifically induced by Shigella within foci of infection.
Fig. 2: Neutrophils are not essential for O2 depletion in infected tissues, which is mainly caused by Shigella aerobic respiration.
Fig. 3: Aerobic respiration is required for hypoxia induction and efficient colonic mucosa colonization by Shigella in vivo.
Fig. 4: S. flexneri T3SS is inactive in the colonic mucosa, supporting foci of infection extension.

Data availability

The datasets generated and/or analysed during the current study are available from the corresponding author on reasonable request.

References

  1. 1.

    Marteyn, B. et al. Modulation of Shigella virulence in response to available oxygen in vivo. Nature 465, 355–358 (2010).

    CAS  Article  Google Scholar 

  2. 2.

    Cramer, T. et al. HIF-1α is essential for myeloid cell-mediated inflammation. Cell 112, 645–657 (2003).

    CAS  Article  Google Scholar 

  3. 3.

    Peyssonnaux, C. et al. HIF-1α expression regulates the bactericidal capacity of phagocytes. J. Clin. Invest. 115, 1806–1815 (2005).

    CAS  Article  Google Scholar 

  4. 4.

    Walmsley, S. R. et al. Hypoxia-induced neutrophil survival is mediated by HIF-1α-dependent NF-κB activity. J. Exp. Med. 201, 105–115 (2005).

    CAS  Article  Google Scholar 

  5. 5.

    Huether, S. E. & McCance, K. L. Understanding Pathophysiology (Elsevier Health Sciences, 2015).

  6. 6.

    Karhausen, J. et al. Epithelial hypoxia-inducible factor-1 is protective in murine experimental colitis. J. Clin. Invest. 114, 1098–1106 (2004).

    CAS  Article  Google Scholar 

  7. 7.

    Campbell, E. L. et al. Transmigrating neutrophils shape the mucosal microenvironment through localized oxygen depletion to influence resolution of inflammation. Immunity 40, 66–77 (2014).

    CAS  Article  Google Scholar 

  8. 8.

    Arena, E. T., Tinevez, J.-Y., Nigro, G., Sansonetti, P. J. & Marteyn, B. S. The infectious hypoxia: occurrence and causes during Shigella infection. Microbes Infect. 19, 157–165 (2017).

    Article  Google Scholar 

  9. 9.

    Ziemer, L. S. et al. Noninvasive imaging of tumor hypoxia in rats using the 2-nitroimidazole 18F-EF5. Eur. J. Nucl. Med. Mol. Imaging 30, 259–266 (2003).

    CAS  Article  Google Scholar 

  10. 10.

    Bumann, D. Heterogeneous host–pathogen encounters: act locally, think globally. Cell Host Microbe 17, 13–19 (2015).

    CAS  Article  Google Scholar 

  11. 11.

    Davis, K. M. & Isberg, R. R. Defining heterogeneity within bacterial populations via single cell approaches. BioEssays 38, 782–790 (2016).

    Article  Google Scholar 

  12. 12.

    Anderson, M. C. et al. MUB40 binds to lactoferrin and stands as a specific neutrophil marker. Cell Chem. Biol. 25, 483–493 (2018).

    CAS  Article  Google Scholar 

  13. 13.

    Sheridan, W. G., Lowndes, R. H. & Young, H. L. Intraoperative tissue oximetry in the human gastrointestinal tract. Am. J. Surg. 159, 314–319 (1990).

    CAS  Article  Google Scholar 

  14. 14.

    Unden, G. & Trageser, M. Oxygen regulated gene expression in Escherichia coli: control of anaerobic respiration by the FNR protein. Antonie Van Leeuwenhoek 59, 65–76 (1991).

    CAS  Article  Google Scholar 

  15. 15.

    Way, S. S., Sallustio, S., Magliozzo, R. S. & Goldberg, M. B. Impact of either elevated or decreased levels of cytochrome bd expression on Shigella flexneri virulence. J. Bacteriol. 181, 1229–1237 (1999).

    CAS  PubMed  PubMed Central  Google Scholar 

  16. 16.

    Campbell-Valois, F.-X. et al. A fluorescent reporter reveals on/off regulation of the Shigella type III secretion apparatus during entry and cell-to-cell spread. Cell Host Microbe 15, 177–189 (2014).

    CAS  Article  Google Scholar 

  17. 17.

    Davis, K. M., Mohammadi, S. & Isberg, R. R. Community behavior and spatial regulation within a bacterial microcolony in deep tissue sites serves to protect against host attack. Cell Host Microbe 17, 21–31 (2015).

    CAS  Article  Google Scholar 

  18. 18.

    Hughes, E. R. et al. Microbial respiration and formate oxidation as metabolic signatures of inflammation-associated dysbiosis. Cell Host Microbe 21, 208–219 (2017).

    CAS  Article  Google Scholar 

  19. 19.

    Nizet, V. & Johnson, R. S. Interdependence of hypoxic and innate immune responses. Nat. Rev. Immunol. 9, 609–617 (2009).

    CAS  Article  Google Scholar 

  20. 20.

    Taylor, C. T. & Colgan, S. P. Regulation of immunity and inflammation by hypoxia in immunological niches. Nat. Rev. Immunol. 17, 774–785 (2017).

    CAS  Article  Google Scholar 

  21. 21.

    Light, S. H. et al. A flavin-based extracellular electron transfer mechanism in diverse Gram-positive bacteria. Nature 562, 140–144 (2018).

    CAS  Article  Google Scholar 

  22. 22.

    Dejean, L., Beauvoit, B., Guérin, B. & Rigoulet, M. Growth of the yeast Saccharomyces cerevisiae on a non-fermentable substrate: control of energetic yield by the amount of mitochondria. Biochim. Biophys. Acta 1457, 45–56 (2000).

    CAS  Article  Google Scholar 

  23. 23.

    Monceaux, V. et al. Anoxia and glucose supplementation preserve neutrophil viability and function. Blood 128, 993–1002 (2016).

    CAS  Article  Google Scholar 

  24. 24.

    Arena, E. T. et al. Bioimage analysis of Shigella infection reveals targeting of colonic crypts. Proc. Natl Acad. Sci. USA 112, E3282–E3290 (2015).

    CAS  Article  Google Scholar 

  25. 25.

    Schindelin, J. et al. Fiji: an open-source platform for biological-image analysis. Nat. Methods 9, 676–682 (2012).

    CAS  Article  Google Scholar 

  26. 26.

    Tinevez, J. Y. et al. TrackMate: an open and extensible platform for single-particle tracking. Methods 115, 80–90 (2017).

    CAS  Article  Google Scholar 

Download references

Acknowledgements

We acknowledge France-BioImaging infrastructure, supported by the French National Research Agency (ANR-10-INBS-04, Imagopole; to J.-Y.T.), ANR JCJC 2017-17-CE15-0012 (to B.S.M.) and the European Research Council (ERC grant 2009-AdG HOMEOPATH; to P.J.S.). E.T.A. was a Pasteur Foundation and Pasteur-Roux fellow.

Author information

Affiliations

Authors

Contributions

B.S.M., J.-Y.T. and E.T.A. designed the experiments, interpreted the data and wrote the paper. M.A. designed the Shigella mutants. G.N., L.I., A.A., M.F. and F.-X.C.-V. contributed to studying the Shigella mutants in vitro and in vivo. J.-Y.T. conducted quantitative analysis of the data. A.D., S.L.S. and P.J.S. contributed to data interpretation.

Corresponding author

Correspondence to Benoit S. Marteyn.

Ethics declarations

Competing interests

The authors declare no competing interests.

Additional information

Publisher’s note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary information

Supplementary Information

Supplementary Figs. 1–5 and Supplementary Tables 1–12.

Reporting Summary

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Tinevez, JY., Arena, E.T., Anderson, M. et al. Shigella-mediated oxygen depletion is essential for intestinal mucosa colonization. Nat Microbiol 4, 2001–2009 (2019). https://doi.org/10.1038/s41564-019-0525-3

Download citation

Further reading

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing