Role of bacterial surface structures on the interaction of Klebsiella pneumoniae with phagocytes

PLoS One. 2013;8(2):e56847. doi: 10.1371/journal.pone.0056847. Epub 2013 Feb 15.

Abstract

Phagocytosis is a key process of the immune system. The human pathogen Klebsiella pneumoniae is a well known example of a pathogen highly resistant to phagocytosis. A wealth of evidence demonstrates that the capsule polysaccharide (CPS) plays a crucial role in resistance to phagocytosis. The amoeba Dictyostelium discoideum shares with mammalian macrophages the ability to phagocytose and kill bacteria. The fact that K. pneumoniae is ubiquitous in nature and, therefore, should avoid predation by amoebae, poses the question whether K. pneumoniae employs similar means to counteract amoebae and mammalian phagocytes. Here we developed an assay to evaluate K. pneumoniae-D. discoideum interaction. The richness of the growth medium affected the threshold at which the cps mutant was permissive for Dictyostelium and only at lower nutrient concentrations the cps mutant was susceptible to predation by amoebae. Given the critical role of bacterial surface elements on host-pathogen interactions, we explored the possible contribution of the lipopolysaccharide (LPS) and outer membrane proteins (OMPs) to combat phagoyctosis by D. discoideum. We uncover that, in addition to the CPS, the LPS O-polysaccharide and the first core sugar participate in Klebsiella resistance to predation by D. discoideum. K. pneumoniae LPS lipid A decorations are also necessary to avoid predation by amoebae although PagP-dependent palmitoylation plays a more important role than the lipid A modification with aminoarabinose. Mutants lacking OMPs OmpA or OmpK36 were also permissive for D. discoideium growth. Except the LPS O-polysaccharide mutants, all mutants were more susceptible to phagocytosis by mouse alveolar macrophages. Finally, we found a correlation between virulence, using the pneumonia mouse model, and resistance to phagocytosis. Altogether, this work reveals novel K. pneumoniae determinants involved in resistance to phagocytosis and supports the notion that Dictyostelium amoebae might be useful as host model to measure K. pneumoniae virulence and not only phagocytosis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Arabinose / analogs & derivatives
  • Arabinose / metabolism
  • Bacterial Outer Membrane Proteins / metabolism
  • Bacterial Proteins / genetics
  • Carbohydrate Sequence
  • Dictyostelium / microbiology
  • Dictyostelium / physiology
  • Female
  • Host-Pathogen Interactions*
  • Klebsiella pneumoniae / genetics
  • Klebsiella pneumoniae / metabolism
  • Klebsiella pneumoniae / pathogenicity
  • Klebsiella pneumoniae / physiology*
  • Lipid A / metabolism
  • Macrophages, Alveolar / cytology
  • Macrophages, Alveolar / microbiology
  • Mice
  • Molecular Sequence Data
  • Mutation
  • O Antigens / chemistry
  • O Antigens / metabolism
  • Palmitic Acid / metabolism
  • Phagocytes / cytology
  • Phagocytes / microbiology*
  • Phagocytosis
  • Porins / genetics

Substances

  • Bacterial Outer Membrane Proteins
  • Bacterial Proteins
  • Lipid A
  • O Antigens
  • OmpK36 protein, Klebsiella pneumoniae
  • Porins
  • aminoarabinose
  • OMPA outer membrane proteins
  • Palmitic Acid
  • Arabinose

Grants and funding

Part of this work was supported by Ministerio de Economa y Competitividad (Spain) and from Generalitat de Catalunya (Centre de Referència en Biotecnologia) grants to J.M.T.; by grants from Biomedicine Program (SAF2009-07885; Ministerio de Economa y Competitividad) and SEPAR (Sociedad Española de Neumologa, project reference 068/2011) to J.A.B. The laboratory of Microbial Pathogenesis is supported by a grant to Competitive groups (project reference 46/2011) from Govern Illes Balears (co-funded by European Regional Development Fund). CIBERES is an initiative from Instituto de Salud Carlos III. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.