Letter | Published:

Listeria monocytogenes exploits efferocytosis to promote cell-to-cell spread

Nature volume 509, pages 230234 (08 May 2014) | Download Citation

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Abstract

Efferocytosis, the process by which dying or dead cells are removed by phagocytosis, has an important role in development, tissue homeostasis and innate immunity1. Efferocytosis is mediated, in part, by receptors that bind to exofacial phosphatidylserine (PS) on cells or cellular debris after loss of plasma membrane asymmetry. Here we show that a bacterial pathogen, Listeria monocytogenes, can exploit efferocytosis to promote cell-to-cell spread during infection. These bacteria can escape the phagosome in host cells by using the pore-forming toxin listeriolysin O (LLO) and two phospholipase C enzymes2. Expression of the cell surface protein ActA allows L. monocytogenes to activate host actin regulatory factors and undergo actin-based motility in the cytosol, eventually leading to formation of actin-rich protrusions at the cell surface. Here we show that protrusion formation is associated with plasma membrane damage due to LLO’s pore-forming activity. LLO also promotes the release of bacteria-containing protrusions from the host cell, generating membrane-derived vesicles with exofacial PS. The PS-binding receptor TIM-4 (encoded by the Timd4 gene) contributes to efficient cell-to-cell spread by L. monocytogenes in macrophages in vitro and growth of these bacteria is impaired in Timd4−/− mice. Thus, L. monocytogenes promotes its dissemination in a host by exploiting efferocytosis. Our results indicate that PS-targeted therapeutics may be useful in the fight against infections by L. monocytogenes and other bacteria that use similar strategies of cell-to-cell spread during infection.

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Acknowledgements

We are grateful to S. Gray-Owen, S. Grinstein and D. Portnoy for providing reagents and advice and to D. Holmyard for help with electron microscopy. J.H.B. holds the Pitblado Chair in Cell Biology. Infrastructure for the Brumell laboratory was provided by a Leader’s Opportunity Fund grant from the Canadian Foundation for Innovation and the Ontario Innovation Trust. R.F. was supported by a postdoctoral fellowship from the Canadian Institutes of Health Research in partnership with the Canadian Association of Gastroenterology and the Crohn’s and Colitis Foundation of Canada. S.O. was supported by a postdoctoral fellowship from the Research Training Committee at the Hospital for Sick Children. This work was supported by an operating grant from The Arthritis Society of Canada (#RG11/013) to J.H.B. and a US Public Health Service grant (AI053669) from the National Institutes of Health to D.E.H.

Author information

Affiliations

  1. Cell Biology Program, Hospital for Sick Children, Toronto, Ontario M5G0A4, Canada

    • Mark A. Czuczman
    • , Ramzi Fattouh
    • , Veronica Canadien
    • , Suzanne Osborne
    • , Aleixo M. Muise
    •  & John H. Brumell
  2. Department of Molecular Genetics, University of Toronto, Toronto, Ontario M5S1A8, Canada

    • Mark A. Czuczman
    •  & John H. Brumell
  3. Department of Cell Biology and Institute of Biomembranes, University Medical Center Utrecht, 3584 CX Utrecht, the Netherlands

    • Jorik M. van Rijn
  4. Division of Gastroenterology, Hepatology, and Nutrition, Department of Paediatrics, Hospital for Sick Children, Toronto, Ontario M5G1X8, Canada

    • Aleixo M. Muise
  5. Institute of Medical Science, University of Toronto, Toronto, Ontario M5S1A8, Canada

    • Aleixo M. Muise
    •  & John H. Brumell
  6. Sickkids IBD Centre, Hospital for Sick Children, Toronto, Ontario M5G1X8, Canada

    • Aleixo M. Muise
    •  & John H. Brumell
  7. Center for Neurologic Diseases, Brigham and Women’s Hospital, Harvard Medical School, Boston, Massachusetts 02115, USA

    • Vijay K. Kuchroo
  8. Department of Microbiology and Immunobiology, Harvard Medical School, Boston, Massachusetts 02115, USA

    • Darren E. Higgins

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Contributions

J.H.B., M.A.C., S.O. and D.E.H. designed the experiments and wrote the paper. M.A.C., R.F., J.M.v.R., V.C. and S.O. performed the experiments. A.M.M. and V.K.K. contributed reagents and consultations.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to John H. Brumell.

Extended data

Supplementary information

Videos

  1. 1.

    Lm protrusion formation leads to exofacial PS exposure on membrane vesicles

    Cells were transfected with LifeAct-RFP (red) and then infected with wild type Lm expressing GFP at an MOI of 100 for 6h. Live infected cells were then analyzed by spinning disk confocal microscopy with Annexin V-Alexa 647 in the medium to label exofacial PS (blue). Frames from this video were cropped and are presented in Figure 3B. Images are representative of 3 independent experiments.

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DOI

https://doi.org/10.1038/nature13168

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