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Dictyostelium Host Response to Legionella Infection: Strategies and Assays

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Legionella

Part of the book series: Methods in Molecular Biology ((MIMB,volume 954))

Abstract

The professional phagocyte Dictyostelium discoideum is a simple eukaryotic microorganism, whose natural habitat is deciduous forest soil and decaying leaves, where the amoebae feed on bacteria and grow as separate, independent, single cells. In the last decade, the organism has been successfully used as a host for several human pathogens, including Legionella pneumophila, Mycobacterium avium and Mycobacterium marinum,Pseudomonas aeruginosa, Klebsiella pneumoniae, Cryptococcus neoformans, and Salmonella typhimurium. To dissect the complex cross-talk between host and pathogen Dictyostelium offers easy cultivation, a high quality genome sequence and excellent molecular genetic and biochemical tools. Dictyostelium cells are also extremely suitable for cell biological studies, which in combination with in vivo expression of fluorescence-tagged proteins allow investigating the dynamics of bacterial uptake and infection. Inactivation of genes by homologous recombination as well as gene rescue and overexpression are well established and a large mutant collection is available at the Dictyostelium stock center, favoring identification of host resistance or susceptibility genes. Here, we briefly introduce the organism, address the value of Dictyostelium as model host, describe strategies to identify host cell factors important for infection followed by protocols for cell culture and storage, uptake and infection, and confocal microscopy of infected cells.

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Acknowledgments

This work was supported by the Region Piemonte (SB), and the Deutsche Forschungsgemeinschaft (SFB 670, Innate Immunity) and Köln Fortune (LE).

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Authors and Affiliations

  1. Department of Clinical and Biological Sciences, University of Turin, Orbassano, Italy

    Salvatore Bozzaro & Barbara Peracino

  2. Medical Faculty, Center for Biochemistry, University of Cologne, Cologne, Germany

    Ludwig Eichinger

Authors
  1. Salvatore Bozzaro
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  2. Barbara Peracino
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  3. Ludwig Eichinger
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Corresponding author

Correspondence to Salvatore Bozzaro .

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Editors and Affiliations

  1. , Biology of Intracellular Bacteria, Institut Pasteur, Rue du Dr. Roux 28, Paris, 75724, France

    Carmen Buchrieser

  2. Max von Pettenkofer Institute, Ludwig-Maximilians-University, Pettenkoferstrasse 9a, Munich, 80336, Germany

    Hubert Hilbi

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Bozzaro, S., Peracino, B., Eichinger, L. (2013). Dictyostelium Host Response to Legionella Infection: Strategies and Assays. In: Buchrieser, C., Hilbi, H. (eds) Legionella. Methods in Molecular Biology, vol 954. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-161-5_26

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  • DOI: https://doi.org/10.1007/978-1-62703-161-5_26

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-160-8

  • Online ISBN: 978-1-62703-161-5

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