Abstract
Listeria monocytogenes is a bacterial pathogen which invades and multiplies within non-professional phagocytes. Signaling cascades involved in cellular entry have been extensively analyzed, but the events leading to vacuolar escape remain less clear. In this chapter, we detail a microscopy FRET-based assay which allows quantitatively measuring L. monocytogenes infection and escape from its internalization vacuole, as well as a correlative light/electron microscopy method to investigate the morphological features of the vacuolar compartments containing L. monocytogenes.
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References
de Noordhout CM, Devleesschauwer B, Angulo F et al (2014) The global burden of listeriosis: a systematic review and meta-analysis. Lancet Infect Dis 14:1073–1082
Mackaness GB (1962) Cellular resistance to infection. J Exp Med 116:381–406
Cossart P (2011) Illuminating the landscape of host-pathogen interactions with the bacterium Listeria monocytogenes. Proc Natl Acad Sci U S A 108:19484–19491. doi:10.1073/pnas.1112371108
Pizarro-Cerdá J, Kühbacher A, Cossart P (2012) Entry of Listeria monocytogenes in mammalian epithelial cells: an updated view. Cold Spring Harb Perspect Med 2:a010009. doi:10.1101/cshperspect.a010009
Kühbacher A, Emmenlauer M, Rämö P et al (2015) Genome-wide siRNA screen identifies complementary signaling pathways involved in Listeria infection and reveals different actin nucleation mechanisms during Listeria cell invasion and actin comet tail formation. MBio 6:e00598–15. doi:10.1128/mBio.00598-15
Bonazzi M, Kühbacher A, Toledo-Arana A et al (2012) A common clathrin-mediated machinery co-ordinates cell–cell adhesion and bacterial internalization. Traffic 13:1653–1666. doi:10.1111/tra.12009
Pizarro-Cerdá J, Bonazzi M, Cossart P (2010) Clathrin-mediated endocytosis: what works for small, also works for big. Bioessays 32:496–504. doi:10.1002/bies.200900172
Pizarro-Cerdá J, Cossart P (2009) Listeria monocytogenes membrane trafficking and lifestyle: the exception or the rule? Annu Rev Cell Dev Biol 25:649–670. doi:10.1146/annurev.cellbio.042308.113331
Ireton K, Payrastre B, Chap H et al (1996) A role for phosphoinositide 3-kinase in bacterial invasion. Science 274:780–782
Kühbacher A, Dambournet D, Echard A et al (2012) Phosphatidylinositol 5-phosphatase oculocerebrorenal syndrome of Lowe protein (OCRL) controls actin dynamics during early steps of Listeria monocytogenes infection. J Biol Chem 287:13128–13136. doi:10.1074/jbc.M111.315788
Pizarro-Cerdá J, Kühbacher A, Cossart P (2014) Phosphoinositides and host-pathogen interactions. Biochim Biophys Acta 1851:911–918. doi:10.1016/j.bbalip.2014.09.011
Seastone CV (1935) Pathogenic organisms of the genus Listerella. J Exp Med 62:203–212
Gaillard JL, Berche P, Mounier J et al (1987) In vitro model of penetration and intracellular growth of Listeria monocytogenes in the human enterocyte-like cell line Caco-2. Infect Immun 55:2822–2829
Henry R, Shaughnessy L, Loessner MJ et al (2006) Cytolysin-dependent delay of vacuole maturation in macrophages infected with Listeria monocytogenes. Cell Microbiol 8:107–119. doi:10.1111/j.1462-5822.2005.00604.x
Mengaud J, Braun-Breton C, Cossart P (1991) Identification of phosphatidylinositol-specific phospholipase C activity in Listeria monocytogenes: a novel type of virulence factor? Mol Microbiol 5:367–372
Vázquez-Boland JA, Kocks C, Dramsi S et al (1992) Nucleotide sequence of the lecithinase operon of Listeria monocytogenes and possible role of lecithinase in cell-to-cell spread. Infect Immun 60:219–230
Barry RA, Bouwer HG, Portnoy DA, Hinrichs DJ (1992) Pathogenicity and immunogenicity of Listeria monocytogenes small-plaque mutants defective for intracellular growth and cell-to-cell spread. Infect Immun 60:1625–1632
Singh R, Jamieson A, Cresswell P (2008) GILT is a critical host factor for Listeria monocytogenes infection. Nature 455:1244–1247. doi:10.1038/nature07344
Radtke AL, Anderson KL, Davis MJ et al (2011) Listeria monocytogenes exploits cystic fibrosis transmembrane conductance regulator (CFTR) to escape the phagosome. Proc Natl Acad Sci U S A 108:1633–1638. doi:10.1073/pnas.1013262108
Davis MJ, Gregorka B, Gestwicki JE, Swanson JA (2012) Inducible renitence limits Listeria monocytogenes escape from vacuoles in macrophages. J Immunol 189:4488–4495. doi:10.4049/jimmunol.1103158
Quereda JJ, Pizarro-Cerdá J, Balestrino D et al (2015) A dual microscopy-based assay to assess Listeria monocytogenes cellular entry and vacuolar escape. Appl Environ Microbiol 82(1):211–217. doi:10.1128/AEM.02302-15
Mellouk N, Weiner A, Aulner N et al (2014) Shigella subverts the host recycling compartment to rupture its vacuole. Cell Host Microbe 16:517–530. doi:10.1016/j.chom.2014.09.005
Mostowy S, Janel S, Forestier C et al (2011) A role for Septins in the interaction between the Listeria monocytogenes invasion protein InlB and the met receptor. Biophys J 100:1949–1959. doi:10.1016/j.bpj.2011.02.040
Quereda JJ, Pucciarelli MG (2014) Deletion of the membrane protein Lmo0412 increases the virulence of Listeria monocytogenes. Microbes Infect 16:623–632. doi:10.1016/j.micinf.2014.07.002
Ripio MT, Domínguez-Bernal G, Lara M et al (1997) A Gly145Ser substitution in the transcriptional activator PrfA causes constitutive overexpression of virulence factors in Listeria monocytogenes. J Bacteriol 179:1533–1540
Kühbacher A, Cossart P, Pizarro-Cerdá J (2014) Internalization assays for Listeria monocytogenes. In: Methods in molecular biology. Springer New York, New York, NY, pp 167–178
Acknowledgments
We thank Servier Medical Art (http://www.servier.com/Powerpoint-image-bank) for providing drawings used in Fig. 1. Work in the laboratory of P.C. is supported by the Institut Pasteur (Unité des Interactions Bactéries-Cellules), the Institut National de la Santé et de la Recherche Médicale (Unité 604), the Institut National de la Recherche Agronomique (Unité Sous Contrat 2020), the Fondation Les Mousquetaires, and an European Research Council Advanced Grant (BacEpiCell 670823). The Imagopole is part of the FranceBioImaging infrastructure supported by the French National Research Agency (ANR-10-INSB-04-01, “Investments for the Future”) and is grateful to supports by Conseil de la Region Ile-de-France (program Sesame 2007, project Imagopole to S.S.) and from the Fondation Française pour la Recherche Médicale (FRM, Programme Grands Equipements to N.A.). J.E. is supported by the Institut Pasteur (PTR-460), by the Institut Pasteur Carnot-MIE program and by a European Research Council Starting Grant (RuptEffects 261166). J.P.C. is supported by the Institut Pasteur (PTR-460 and PTR-521) and by the French National Research Agency (ANR-15-CE15-0017). P.C. and J.E. acknowledge the LabEx consortium IBEID. P.C. is a Howard Hughes Medical Institute Senior International Research Scholar.
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Quereda, J.J. et al. (2017). Assessing Vacuolar Escape of Listeria Monocytogenes . In: Nordenfelt, P., Collin, M. (eds) Bacterial Pathogenesis. Methods in Molecular Biology, vol 1535. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6673-8_11
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DOI: https://doi.org/10.1007/978-1-4939-6673-8_11
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