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Listeriolysin O allows Listeria monocytogenes replication in macrophage vacuoles

Nature volume 451, pages 350–354 (2008)Cite this article

Abstract

Listeria monocytogenes is an intracellular bacterial pathogen that replicates rapidly in the cytosol of host cells during acute infection1. Surprisingly, these bacteria were found to occupy vacuoles in liver granuloma macrophages during persistent infection of severe combined immunodeficient (SCID) mice2. Here we show that L. monocytogenes can replicate in vacuoles within macrophages. In livers of SCID mice infected for 21 days, we observed bacteria in large LAMP1+ compartments that we termed spacious Listeria-containing phagosomes (SLAPs). SLAPs were also observed in vitro, and were found to be non-acidic and non-degradative compartments that are generated in an autophagy-dependent manner. The replication rate of bacteria in SLAPs was found to be reduced compared to the rate of those in the cytosol. Listeriolysin O (LLO, encoded by hly), a pore-forming toxin essential for L. monocytogenes virulence1, was necessary and sufficient for SLAP formation. A L. monocytogenes mutant with low LLO expression was impaired for phagosome escape but replicated slowly in SLAPs over a 72 h period. Therefore, our studies reveal a role for LLO in promoting L. monocytogenes replication in vacuoles and suggest a mechanism by which this pathogen can establish persistent infection in host macrophages.

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Figure 1: L. monocytogenes colonize SLAPs during chronic infection of SCID mice.
Figure 2: L. monocytogenes replicate slowly in SLAPs during in vitro infection of macrophages.
Figure 3: SLAP formation requires bacterial LLO expression.
Figure 4: Impaired LLO expression allows slow bacterial replication within vacuoles.

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Acknowledgements

J.H.B. holds an Investigators in Pathogenesis of Infectious Disease Award from the Burroughs Wellcome Fund and is the recipient of the Premier’s Research Excellence Award from the Ontario Ministry of Economic Development and Trade and the Boehringer Ingelheim (Canada) Young Investigator Award in Biological Sciences. Laboratory infrastructure was provided by a New Opportunities Fund from the Canadian Foundation for Innovation and the Ontario Innovation Trust. C.L.B. holds a Canada Graduate Scholarship from the Natural Sciences and Engineering Research Council of Canada. N.A.K. holds a CAG/CIHR/Axcan Pharma fellowship from the Canadian Association of Gastroenterology. B.E.S. is supported by Canadian Institutes of Health Research and McLaughlin Centre for Molecular Medicine MD/PhD studentships. We are grateful to E. R. Unanue for performing in vivo infections of mice and providing tissue sections and electron micrographs. We thank D. Brown, P. Cossart, J. Danska, E. Gouin, S. Grinstein, N. Jones, N. Mizushima, D. Portnoy, and T. Yoshimori for providing reagents and suggestions. We also thank M. Woodside, P. Paroutis and R. Temkin for assistance with microscopy.

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Authors and Affiliations

  1. Cell Biology Program, Hospital for Sick Children, Toronto, Ontario M5G 1X8, Canada ,

    Cheryl L. Birmingham, Veronica Canadien, Natalia A. Kaniuk, Benjamin E. Steinberg & John H. Brumell

  2. Department of Molecular Genetics,,

    Cheryl L. Birmingham & John H. Brumell

  3. Institute of Medical Science, University of Toronto, Toronto, Ontario M5S 1A8, Canada ,

    Benjamin E. Steinberg & John H. Brumell

  4. Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, Massachusetts 02115-6092, USA,

    Darren E. Higgins

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Correspondence to John H. Brumell.

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Birmingham, C., Canadien, V., Kaniuk, N. et al. Listeriolysin O allows Listeria monocytogenes replication in macrophage vacuoles. Nature 451, 350–354 (2008). https://doi.org/10.1038/nature06479

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