The soil amoeba Dictyostelium discoideum is a haploid eukaryote that, upon starvation, aggregates and enters a developmental cycle to produce fruiting bodies. In this study, we infected single‐cell stages of D. discoideum with different Legionella species. Intracellular growth of Legionella in this new host system was compared with their growth in the natural host Acanthamoeba castellanii. Transmission electron microscopy of infected D. discoideum cells revealed that legionellae reside within the phagosome. Using confocal microscopy, it was observed that replicating, intracellular, green fluorescent protein (GFP)‐tagged legionellae rarely co‐localized with fluorescent antibodies directed against the lysosomal protein DdLIMP of D. discoideum. This indicates that the bacteria inhibit the fusion of phagosomes and lysosomes in this particular host system. In addition, Legionella infection of D. discoideum inhibited the differentiation of the host into the multicellular fruiting stage. Co‐culture studies with profilin‐minus D. discoideum mutants and Legionella resulted in higher rates of infection when compared with infections of wild‐type amoebae. Because the amoebae are amenable to genetic manipulation as a result of their haploid genome and because a number of cellular markers are available, we show for the first time that D. discoideum is a valuable model system for studying intracellular pathogenesis of microbial pathogens.
Legionella pneumophila is a facultative intracellular parasite able to replicate within and to kill a variety of eukaryotic cells. One possible killing mechanism is the induction of programmed cell death. Based on electron microscopy and flow cytometry studies using the phosphatidylserine binding protein annexin V, we could demonstrate that L. pneumophila is able to induce apoptosis in human monocytes which was clearly dependent on the multiplicity of infection, the time postinfection and the intracellular location of the bacteria. Furthermore, it became evident that Legionella‐induced apoptosis does not require the TNF‐α mediated signal‐transduction pathway. By studying infection in Acanthamoeba castellanii, we found that L. pneumophila is not able to induce programmed cell death in their natural host cells indicating that different mechanisms are responsible for host cell killing in protozoan and human cells.
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