Areas of Interest
Legionella pneumophila growth in macrophages
Innate and adaptive immune responses are initiated when macrophages ingest microbes. To investigate the mechanisms that govern the outcome of this encounter, we exploit a bacterial pathogen as a genetic probe of macrophage function. Legionella pneumophila is an opportunistic human pathogen whose natural reservoir is fresh water amoebae. When inhaled, the gram-negative bacteria can colonize alveolar macrophages and cause the severe pneumonia, Legionnaires' disease.
Metabolic cues govern virulence expression. To persist in the environment, L. pneumophilaalternates between distinct cell types. A replicative cell grows within vacuoles of amoebae and macrophages, and a motile and resilient transmissive form is equipped to escape a spent host and primed to invade a naive one. By applying genetic, biochemical and cell biological methods, we have identified a variety of metabolic cues that govern the pathogen's lifecycle. To resume replication, intracellular L. pneumophila rely on Phagosomal transporter proteins to obtain essential metabolites and the bacterial enzyme SpoT to degrade the alarmone guanosine tetraphosphate (ppGpp). Once the intracellular progeny have exhausted the local nutrient supply, ppGpp accumulates and cooperates with other regulatory proteins to coordinate bacterial expression of transmissive traits, including cytotoxicity, motility, stress resistance, and the capacity to block phagosome-lysosome fusion. By coupling cellular differentiation to its metabolic state, L. pneumophila swiftly acclimates to stresses encountered in its host or the environment, thereby enhancing its overall fitness.
Autophagy and pyroptosis, two barriers to infection. Although L. pneumophila can replicate in human macrophages and fresh water amoebae, mice are naturally resistant to infection. Accordingly, our laboratory exploits a mouse infection model to investigate how the host innate immune system can detect and respond to infection by intracellular pathogens. Genetic analysis of human Crohn's disease, the plant response to tobacco mosaic virus, and mouse restriction of L. pneumophila infection each indicate that cells coordinately regulate autophagy and programmed cell death to combat infection. Accordingly, we are applying bacterial and mouse genetics and cell biological methods to test the hypothesis that, in response to cytosolic contamination with flagellin or other microbial products, NOD-like receptor proteins equip mouse macrophages either to induce autophagy to degrade intracellular microbes or to undergo pyroptosis, a failsafe caspase-1-dependent pro-inflammatory cell death.
Dalebroux, Z. D. and M. S. Swanson: ppGpp magic beyond RNA polymerase. Nat. Rev. Microbiol.,10:203-212, 2012.
Swanson, M: Autophagy as a barrier to infection, in Gordon, S. (ed.), Cells of the Innate Immune System: Roles in health and disease, The Biomedical & Life Sciences Collection, Henry Stewart Talks Ltd, London, 2012
Bryan, A. and M. S. Swanson: Oligonucleotides stimulate genomic alterations of Legionella pneumophila. Mol. Microbiol., 80, 231-247, 2011.
Joshi, A. D. and M. S. Swanson: Secrets of a successful pathogen: Legionella resistance to progression along the autophagic pathway. Front. Microbio. 2:138, 2011.
Dalebroux, Z. D., S. L. Svensson, E. C. Gaynor, and M. S. Swanson:. ppGpp conjures bacterial virulence. Microbiol. Mol. Bio. Rev. 74, 171-199, 2010.
Dalebroux, Z. D., B. F. Yagi, T. Sahr, C. Buchreiser, and M. S. Swanson: Distinct roles of ppGpp and DksA in Legionella pneumophila differentiation. Mol. Microbiol. 76, 200-219, 2010.
Edwards, R.L., M. Jules, C. Buchrieser and M. S. Swanson: The multi-step design of the LetA/LetS two-component system increases Legionella pneumophila versatility. Infect. Immun. 78, 2571-2583, 2010.
Molofsky, A. B., B. G. Byrne, N. N. Whitfield, C. A. Madigan, E. T. Fuse, K. Tateda, and M. S. Swanson: Cytosolic recognition of flagellin by murine macrophages restricts Legionella pneumophila infection. J. Exp. Med. 203:1093-104, 2006.