title: Perforin-2 Protects Host Cells and Mice by Restricting the Vacuole to Cytosol Transitioning of a Bacterial Pathogen.
abstract: The host-encoded Perforin-2 (encoded by the macrophage-expressed gene 1, Mpeg1), which possesses a pore-forming MACPF domain, reduces the viability of bacterial pathogens that reside within membrane-bound compartments. Here, it is shown that Perforin-2 also restricts the proliferation of the intracytosolic pathogen Listeria monocytogenes Within a few hours of systemic infection, the massive proliferation of L. monocytogenes in Perforin-2(-/-)mice leads to a rapid appearance of acute disease symptoms. We go on to show in cultured Perforin-2(-/-)cells that the vacuole-to-cytosol transitioning of L. monocytogenesis greatly accelerated. Unexpectedly, we found that in Perforin-2(-/-)macrophages,Listeria-containing vacuoles quickly (≤ 15 min) acidify, and that this was coincident with greater virulence gene expression, likely accounting for the more rapid translocation of L. monocytogenes to its replicative niche in the cytosol. This hypothesis was supported by our finding that aL. monocytogenes strain expressing virulence factors at a constitutively high level replicated equally well in Perforin-2(+/+)and Perforin-2(-/-)macrophages. Our findings suggest that the protective role of Perforin-2 against listeriosis is based on it limiting the intracellular replication of the pathogen. This cellular activity of Perforin-2 may derive from it regulating the acidification of Listeria-containing vacuoles, thereby depriving the pathogen of favorable intracellular conditions that promote its virulence gene activity.
authors.affiliation: Department of Microbiology & Immunology, University of Miami Miller School of Medicine, Miami, Florida, USA.
authors.forename: Ryan
authors.initials: R
authors.lastname: McCormack
authors.affiliation: Department of Microbiology & Immunology, University of Miami Miller School of Medicine, Miami, Florida, USA.
authors.forename: Wael
authors.initials: W
authors.lastname: Bahnan
authors.affiliation: Department of Microbiology & Immunology, University of Miami Miller School of Medicine, Miami, Florida, USA.
authors.forename: Niraj
authors.initials: N
authors.lastname: Shrestha
authors.affiliation: Department of Microbiology & Immunology, University of Miami Miller School of Medicine, Miami, Florida, USA.
authors.forename: Justin
authors.initials: J
authors.lastname: Boucher
authors.affiliation: Department of Microbiology & Immunology, University of Miami Miller School of Medicine, Miami, Florida, USA.
authors.forename: Marcella
authors.initials: M
authors.lastname: Barreto
authors.affiliation: Department of Biomedical Engineering, University of Miami, Coral Gables, Florida, USA.
authors.forename: Carlos M
authors.initials: CM
authors.lastname: Barrera
authors.affiliation: Department of Biomedical Engineering, University of Miami, Coral Gables, Florida, USA.
authors.forename: Edward A
authors.initials: EA
authors.lastname: Dauer
authors.affiliation: Department of Microbiology & Immunology, University of Illinois at Chicago, Chicago, Illinois, USA.
authors.forename: Nancy E
authors.initials: NE
authors.lastname: Freitag
authors.affiliation: Department of Microbiology & Immunology, University of Miami Miller School of Medicine, Miami, Florida, USA.
authors.forename: Wasif N
authors.initials: WN
authors.lastname: Khan
authors.affiliation: Department of Microbiology & Immunology, University of Miami Miller School of Medicine, Miami, Florida, USA.
authors.forename: Eckhard R
authors.initials: ER
authors.lastname: Podack
authors.affiliation: Department of Microbiology & Immunology, University of Miami Miller School of Medicine, Miami, Florida, USA kschesser@med.miami.edu.
authors.forename: Kurt
authors.identifier: http://orcid.org/0000-0003-0515-3850
authors.initials: K
authors.lastname: Schesser
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D003600:Cytosol / Q000382:microbiology / Q000502:physiology*; D005786:Gene Expression Regulation / Q000502:physiology; D008089:Listeria monocytogenes / Q000502:physiology* / Q000648:ultrastructure; D008088:Listeriosis / Q000378:metabolism / Q000382:microbiology; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D052899:Pore Forming Cytotoxic Proteins / Q000235:genetics / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D014617:Vacuoles / Q000502:physiology*
chemical_list: D008565:Membrane Proteins; C483884:Mpeg1 protein, mouse; D052899:Pore Forming Cytotoxic Proteins; C510469:perforin 2
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