Source: http://www.asmscience.org/content/book/10.1128/9781555816650.ch32
Timestamp: 2019-04-21 08:51:09+00:00

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Many of the proteins thought to play roles in phagocytic processes in higher eukaryotes have been analyzed in Dictyostelium discoideum through the use of directed deletion mutations. During phagocytosis of either nonpathogens or particles, internalization of the target by D. discoideum proceeds via a process that is very reminiscent of phagocytic uptake by macrophages. The original attempt to isolate D. discoideum phagocytosis-defective mutants that employed a selection strategy was one that was published more than 25 years ago, although the nature of the mutations isolated in this study has never been worked out. There are number of microbial pathogens discussed in this chapter, however, that are documented to interact with amoebae in the environment, making them attractive microorganisms to study with respect to D. discoideum. As described in the discussion of the interaction of Mycobacteria with amoebae, the resemblance of pathogenic mycobacterial species with common environmental isolates suggests that selection for virulence could be partially explained by acquiring resistance to killing by soil amoebae. The soil fungus Cryptococcus neoformans is an opportunistic pathogen of humans that mainly causes disease in immunecompromised patients, such as those with a genetically defective immune system, organ transplant patients on immunosuppressive therapy, and individuals with human immunodeficiency virus infection. There are a number of surrogate hosts that have been used to simplify analysis of virulence factors of pathogens that attack mammalian hosts, most notably Caenorhabditis elegans, Drosophila melanogaster, and Arabidopsis thaliana.
A D. discoideum rtoA − mutant is inefficient at maintaining an L. pneumophila replication compartment. L. pneumophila was incubated with either (A) the wild-type AX4 strain of D. discoideum or (B) an AX4 rtoA − strain for 6 h, fixed, and prepared for electron microscopy. The wild-type strain is found within a spacious vacuole, whereas the rtoA − mutant appears to be in a vacuole that has fused with other compartments in the amoebae. Micrograph is from Li et al. (2005) .
M. marinum resides in two different locales within D. discoideum. Micrographs from (A) Skriwan et al. (2002) and (B) Solomon et al. (2003) . Amoebae were incubated with M. marinum for 24 h, fixed, and processed for transmission electron microscopy. (A) Groups of bacteria found in a single vacuole. (B) Bacteria apparently found in the amoebal cytoplasm. Bacteria free in cytoplasm were originally described as being in individual phagosomes ( Skriwan et al., 2002 ; Solomon et al., 2003 ), but more recent work suggests that the M. marinum may not be in membrane-bound compartments in panel B ( Stamm et al., 2003 ; Hagedorn and Soldati, 2007 ).
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