Interaction between innate immune cells and a bacterial type III secretion system in mutualistic and pathogenic associations

Proc Natl Acad Sci U S A. 2007 May 29;104(22):9481-6. doi: 10.1073/pnas.0700286104. Epub 2007 May 17.

Abstract

Animals house a community of bacterial symbionts in their digestive tracts that contribute to their well being. The medicinal leech, Hirudo verbana, has a remarkably simple gut population carrying two extracellular microbes in the crop where the ingested blood is stored. This simplicity renders it attractive for studying colonization factors. Aeromonas veronii, one of the leech symbionts, can be genetically manipulated and is a pathogen of mammals. Screening transposon mutants of A. veronii for colonization defects in the leech, we found one mutant, JG752, with a transposon insertion in an ascU homolog, encoding an essential component of type III secretion systems (T3SS). Competing JG752 against the wild type revealed that JG752 was increasingly attenuated over time (10-fold at 18 h and >10,000-fold at 96 h). This colonization defect was linked to ascU by complementing JG752 with the operon containing ascU. Fluorescence in situ hybridization analysis revealed that at 42 h 38% of JG752 cells were phagocytosed by leech macrophage-like cells compared with <0.1% of the parental strain. Using mammalian macrophages, a lactate dehydrogenase release assay revealed that cytotoxicity was significantly reduced in macrophages exposed to JG752. In a mouse septicemia model, JG752 killed only 30% of mice, whereas the parent strain killed 100%, showing the importance of T3SS for both pathogenesis and mutualism. Phagocytic immune cells are important not only in defending against pathogens but also in maintaining the mutualistic symbiont community inside the leech, demonstrating that animals use similar, conserved mechanisms to control bacterial populations, even when the outcomes differ dramatically.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Aeromonas / genetics
  • Aeromonas / immunology
  • Aeromonas / metabolism*
  • Aeromonas / pathogenicity*
  • Animals
  • Bacterial Proteins / genetics*
  • Bacterial Proteins / immunology*
  • Bacterial Proteins / metabolism
  • Immunity, Innate / immunology*
  • Mice
  • Molecular Sequence Data
  • Mutation / genetics

Substances

  • Bacterial Proteins

Associated data

  • GENBANK/EF215451