title: Stimulation of autophagy suppresses the intracellular survival of Burkholderia pseudomallei in mammalian cell lines.
abstract: Burkholderia pseudomallei is the causative agent of melioidosis, a tropical infection of humans and other animals. The bacterium is an intracellular pathogen that can escape from endosomes into the host cytoplasm, where it replicates and infects adjacent cells. We investigated the role played by autophagy in the intracellular survival of B. pseudomallei in phagocytic and non-phagocytic cell lines. Autophagy was induced in response to B. pseudomallei invasion of murine macrophage (RAW 264.7) cells and a proportion of the bacteria co-localized with the autophagy effector protein LC3, a marker for autophagosome formation. Pharmacological stimulation of autophagy in RAW 264.7 and murine embryonic fibroblast (MEF) cell lines resulted in increased co-localization of B. pseudomallei with LC3 while basal levels of co-localization could be abrogated using inhibitors of the autophagic pathway. Furthermore, induction of autophagy decreased the intracellular survival of B. pseudomallei in these cell lines, but bacterial survival was not affected in MEF cell lines deficient in autophagy. Treatment of infected macrophages with chloramphenicol increased the proportion of bacteria within autophagosomes indicating that autophagic evasion is an active process relying on bacterial protein synthesis. Consistent with this hypothesis, we identified a B. pseudomallei type III secreted protein, BopA, which plays a role in mediating bacterial evasion of autophagy. We conclude that the autophagic pathway is a component of the innate defense system against invading B. pseudomallei, but which the bacteria can actively evade. However, when autophagy is pharmacologically induced using rapamycin, bacteria are actively sequestered in autophagosomes, ultimately decreasing their survival.
authors.affiliation: Australian Bacterial Pathogenesis Program, Monash University, Clayton campus, Victoria, Australia.
authors.forename: Méabh
authors.initials: M
authors.lastname: Cullinane
authors.forename: Lan
authors.initials: L
authors.lastname: Gong
authors.forename: Xuelei
authors.initials: X
authors.lastname: Li
authors.forename: Natalie
authors.initials: N
authors.lastname: Lazar-Adler
authors.forename: Thien
authors.initials: T
authors.lastname: Tra
authors.forename: Ernst
authors.initials: E
authors.lastname: Wolvetang
authors.forename: Mark
authors.initials: M
authors.lastname: Prescott
authors.forename: John D
authors.initials: JD
authors.lastname: Boyce
authors.forename: Rodney J
authors.initials: RJ
authors.lastname: Devenish
authors.forename: Ben
authors.initials: B
authors.lastname: Adler
mesh_terms: D000730:Androstadienes / Q000494:pharmacology; D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001343:Autophagy / Q000187:drug effects / Q000502:physiology*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D015415:Biomarkers / Q000378:metabolism; D016957:Burkholderia pseudomallei / Q000166:cytology / Q000472:pathogenicity / Q000502:physiology*; D002460:Cell Line / Q000382:microbiology*; D002470:Cell Survival; D006801:Humans; D051379:Mice; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D020123:Sirolimus / Q000494:pharmacology; D000077191:Wortmannin
chemical_list: D000730:Androstadienes; D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D015415:Biomarkers; D047428:Protein Kinase Inhibitors; D011993:Recombinant Fusion Proteins; D020123:Sirolimus; D000077191:Wortmannin