Reduced toxicity of lipo-oligosaccharide from a phoP mutant of Neisseria meningitidis: an in vitro demonstration.
PhoP is part of a two-component regulatory system, which we have previously demonstrated in Neisseria meningitidis and shown to be an important regulator of virulence in an in vivo model. The phoP mutant clearly induced cross-species reactive antibodies and lacks the obvious toxic effects of the wild-type strain. In the current study, we demonstrate distinct differences between the wild-type and mutant strains in an in vitro model of toxicity. At concentrations likely to be present early in an infection, the mutant was more efficient at stimulating an inflammatory response than the wild-type. However, at the concentrations likely to be found at the site of a fulminant infection, the mutant showed significantly weaker ability to stimulate the release of pro-inflammatory cytokines and the production of reactive oxygen and nitrogen intermediates. SDS-PAGE analysis of the isolated LOS from the wild-type and mutant showed a difference in the level of expression of two major species of LOS, a finding which was supported by preliminary MALDI-TOF analysis. These results suggest that the altered toxicity of the mutant may be due to the increased expression of a conformationally altered LOS species, which shows less affinity and avidity for the cellular receptors responsible for the inflammatory response to endotoxin.
Rustam
Tarick
T
Institute for Biological and Biomolecular Science, University of Portsmouth, Hampshire, UK.
McClean
Stephen
S
Newcombe
Jane
J
McFadden
Johnjoe
J
Eales-Reynolds
Lesley-Jane
LJ
D001426:Bacterial Proteins / Q000235:genetics*; D004591:Electrophoresis, Polyacrylamide Gel; D005609:Free Radicals / Q000378:metabolism; D006801:Humans; D066298:In Vitro Techniques; D008070:Lipopolysaccharides / Q000378:metabolism / Q000633:toxicity*; D009000:Monocytes / Q000378:metabolism; D009154:Mutation / Q000502:physiology*; D009345:Neisseria meningitidis / Q000235:genetics* / Q000378:metabolism*; D009584:Nitrogen / Q000378:metabolism; D010957:Plasmids / Q000235:genetics; D017382:Reactive Oxygen Species / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
D001426:Bacterial Proteins; D005609:Free Radicals; D008070:Lipopolysaccharides; D017382:Reactive Oxygen Species; D014409:Tumor Necrosis Factor-alpha; C061064:PhoP protein, Bacteria; D009584:Nitrogen