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.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Saxe, C. L. (1999) Learning from the slime mold: Dictyostelium discoideum and human disease. Am. J. Hum. Genet. 65, 25–30.
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.
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.
Steinert, M., and Heuner, K. (2005) Dictyostelium as host for pathogenesis. Cell. Microbiol. 7, 307–314.
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.
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.
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.
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.
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.
Lu, H., and Clarke, M. (2005) Dynamic properties of Legionella-containing phagosomes in Dictyostelium amoebae. Cell. Microbiol. 7, 995–1007.
Hilbi, H., Segal, G., and Shuman, H. (2001) Icm/Dot-dependent upregulation of phagocytosis by Legionella pneumophila. Mol. Microbiol. 42, 603–617.
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.
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.
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.
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.
Pope, C. D., Dhand, L., and Cianciotto, N. P. (1994) Random mutagenesis of Legionella pneumophila with mini-Tn10. FEMS Microbiol. Lett. 124, 107–111.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2009 Humana Press, a part of Springer Science+Business Media, LLC
About this protocol
Cite this protocol
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
Download citation
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