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Screening of Virulence Traits in Legionella pneumophila and Analysis of the Host Susceptibility to Infection by Using the Dictyostelium Host Model System

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Host-Pathogen Interactions

Part of the book series: Methods in Molecular Biology ((MIMB,volume 470))

Abstract

The social soil amoeba Dictyostelium discoideum has been established as a host model for several human pathogens including Legionella pneumophila. The complete genome sequence, the genetic tractability, and the phagocytic characteristics of Dictyostelium generate many opportunities for the study of host-pathogen interactions. Important applications of this haploid model organism are (i) the use of Dictyostelium cells as a screening system for bacterial virulence, (ii) the use of Dictyostelium mutant cells to identify genetic host determinants of susceptibility and resistance to infection, and (iii) experiments that allow the dissection of the complex cross-talk with infectious agents. Accordingly, this chapter describes a plaque assay to identify attenuated pathogens, an infection assay for the analysis of host cell mutants and pathogens, and a screening method for the isolation of Legionella mutants that are defective in the reprogramming of the phagolysosomal maturation of the host.

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References

  1. Saxe, C. L. (1999) Learning from the slime mold: Dictyostelium discoideum and human disease. Am. J. Hum. Genet. 65, 25–30.

    Article  CAS  PubMed  Google Scholar 

  2. Farbrother, P., Wagner, C., Na, J., Tunggal, B., Morio, T., Urushihara, H., Tanaka, Y., Schleicher, M., Steinert, M., and Eichinger, L. (2006) Dictyostelium transcriptional host cell response upon infection with Legionella. Cell. Microbiol. 8, 438–456.

    Article  CAS  PubMed  Google Scholar 

  3. Solomon, J. M., and Isberg, R. R. (2000) Growth of Legionella pneumophila in Dictyostelium discoideum: a novel system for genetic analysis of host-pathogen interactions. Trends Microbiol. 10, 478–480.

    Article  Google Scholar 

  4. Steinert, M., and Heuner, K. (2005) Dictyostelium as host for pathogenesis. Cell. Microbiol. 7, 307–314.

    Article  CAS  PubMed  Google Scholar 

  5. Cosson, P., Zulianello, L., Join-Lambert, O., Faurisson, F., Gebbie, L., Benghezal, M., van Delden, C., Kocjancic Curty, L. K., and Köhler, T. (2002) Pseudomonas aeruginosa virulence analysed in a Dictyostelium discoideum host system. J. Bacteriol. 184, 3027–3033.

    Article  CAS  PubMed  Google Scholar 

  6. Pukatzki, S., Ma, A. T., Sturtevant, D., Krastins, B., Saracino, D., Nelson, W. C., Heidelberg, J. F., and Mekalanos, J. (2006) Identification of a conserved bacterial protein secretion system in Vibrio cholerae using the Dictyostelium host model system. Proc. Natl. Acad. Sci. 103, 1528–1533.

    Article  CAS  PubMed  Google Scholar 

  7. Skriwan, C., Fajardo, M., Hägele, S., Horn, M., Wagner, M., Michel, R., Krohne, G., Schleicher, M., Hacker, J., and Steinert, M. (2002) Various bacterial pathogens and symbionts infect the amoeba Dictyostelium discoideum. Int. J. Med. Micobiol. 291, 615–624.

    Article  Google Scholar 

  8. Solomon, J. M., Leung, G. S., and Isberg, R. R. (2003) Intracellular replication of Mycobacterium marinum within Dictyostelium discoideum: efficient replication in the absence of host coronin. Infect. Immun. 71, 3578–3586.

    Article  CAS  PubMed  Google Scholar 

  9. Steenbergen, J. N., Nosanchuk, J. D., Malliaris, S. D., and Casadevall, A. (2003) Cryptococcus neoformans virulence is enhanced after growth in the genetically malleable host Dictyostelium discoideum. Infect. Immun. 71, 4862–4872.

    Article  CAS  PubMed  Google Scholar 

  10. Lu, H., and Clarke, M. (2005) Dynamic properties of Legionella-containing phagosomes in Dictyostelium amoebae. Cell. Microbiol. 7, 995–1007.

    Article  CAS  PubMed  Google Scholar 

  11. Hilbi, H., Segal, G., and Shuman, H. (2001) Icm/Dot-dependent upregulation of phagocytosis by Legionella pneumophila. Mol. Microbiol. 42, 603–617.

    Article  CAS  PubMed  Google Scholar 

  12. Fajardo, M., Schleicher, M., Noegel, A., Bozzaro, S., Killinger, S., Heuner, K., Hacker, J., and Steinert, M. (2004) Calnexin, calreticulin and cytoskeleton associated proteins modulate uptake and growth of Legionella pneumophila in Dictyostelium discoideum. Microbiology 150, 2825–2835.

    Article  CAS  PubMed  Google Scholar 

  13. Hägele, S., Köhler, R., Merkert, H., Schleicher, M., Hacker, J., and Steinert, M. (2000) Dictyostelium discoideum: a new host model system for intracellular pathogens of the genus Legionella. Cell. Microbiol. 2, 165–171.

    Article  PubMed  Google Scholar 

  14. Li, Z., Solomon, J. M., and Isberg, R. R. (2005) Dictyostelium discoideum strains lacking the RtoA protein are detective for maturation of the Legionella pneumophila replication vacuole. Cell. Microbiol. 7, 431–442.

    Article  CAS  PubMed  Google Scholar 

  15. Peracino, B., Wagner, C., Balest, A., Balbo, A., Pergolizzi, B., Noegel, A. A., Steinert, M., and Bozzaro, S. (2006) Function and mechanism of action of Dictyostelium Nramp1 (Slc11a1) in bacterial infection. Traffic 7, 22–38.

    Article  CAS  PubMed  Google Scholar 

  16. Pope, C. D., Dhand, L., and Cianciotto, N. P. (1994) Random mutagenesis of Legionella pneumophila with mini-Tn10. FEMS Microbiol. Lett. 124, 107–111.

    Article  CAS  PubMed  Google Scholar 

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© 2009 Humana Press, a part of Springer Science+Business Media, LLC

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Shevchuk, O., Steinert, M. (2009). Screening of Virulence Traits in Legionella pneumophila and Analysis of the Host Susceptibility to Infection by Using the Dictyostelium Host Model System. In: Rupp, S., Sohn, K. (eds) Host-Pathogen Interactions. Methods in Molecular Biology, vol 470. Humana Press. https://doi.org/10.1007/978-1-59745-204-5_4

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  • DOI: https://doi.org/10.1007/978-1-59745-204-5_4

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-886-7

  • Online ISBN: 978-1-59745-204-5

  • eBook Packages: Springer Protocols

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