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Legionella pp 347-370 | Cite as

Dictyostelium Host Response to Legionella Infection: Strategies and Assays

  • Salvatore BozzaroEmail author
  • Simona Buracco
  • Barbara Peracino
  • Ludwig EichingerEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1921)

Abstract

The professional phagocyte Dictyostelium discoideum is a well-established model organism to study host-pathogen interactions. Dictyostelium amoebae grow as separate, independent cells; they divide by binary fission and take up bacteria and yeast via phagocytosis. In the year 2000, D. discoideum was described by two groups as a novel system for genetic analysis of host-pathogen interactions for the intracellular pathogen Legionella pneumophila. Since then additional microbial pathogens that can be studied in D. discoideum have been reported. The organism has various advantages for the dissection of the complex cross-talk between a host and a pathogen. A fully sequenced and well-curated genome is available, there are excellent molecular genetic tools on the market, and the generation of targeted multiple gene knock-outs as well as the realization of untargeted genetic screens is generally straightforward. Dictyostelium also offers easy cultivation, and the cells are suitable for cell biological studies, which in combination with in vivo expression of fluorescence-tagged proteins allows the investigation of the dynamics of bacterial uptake and infection. Furthermore, a large mutant collection is available at the Dictyostelium stock center, favoring the identification of host resistance or susceptibility genes. Here, we briefly describe strategies to identify host cell factors important during an infection, followed by protocols for cell culture and storage, uptake and infection, and confocal microscopy of infected cells.

Key words

Dictyostelium Legionella Amoeba Phagocytosis Macropinocytosis Host-pathogen interaction Uptake assay Infection Confocal microscopy Flow cytometry 

Notes

Acknowledgments

This work was supported by the Compagnia San Paolo (12-CSP-C03-065) (SB), the Deutsche Forschungsgemeinschaft (TP01, SFB 670, Innate Immunity), and Köln Fortune (LE).

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Clinical and Biological SciencesUniversity of TurinOrbassanoItaly
  2. 2.Center for Biochemistry, Medical FacultyUniversity of CologneCologneGermany

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