title: Lipopolysaccharide isolated from Porphyromonas gingivalis grown in hemin-limited chemostat conditions has a reduced capacity for human neutrophil priming.
abstract: One way prokaryotes respond to environmental stresses is by modifying selected outer membrane components. Iron, in the form of hemin, has been shown to be a significant regulator of Porphyromonas gingivalis growth and virulence and of the expression of outer membrane proteins and lipopoly saccharide. Since lipopoly saccharide has profound effects on host immune cells, this study compared the effect of hemin-restricted and hemin-normal P. gingivalis growth conditions on lipopolysaccharide priming of N-formylmethionyl-leucyl-phenylalanine-induced superoxide generation by human neutrophils. P. gingivalis was grown in a chemostat under normal (5 micrograms hemin/ml) and hemin-restricted (0.08 microgram hemin/ml) conditions. Purified lipopolysaccharide from both P. gingivalis normal and hemin-limited environments increased N-formylmethionyl-leucyl-phenylalanine-induced superoxide release by neutrophils in a dose-dependent manner. Lipopolysaccharide isolated from the hemin-normal conditions was a significantly more potent neutrophil priming agent than the lipopolysaccharide isolated from hemin-restricted conditions. Addition of normal human serum enhanced the priming effect of both lipopolysaccharide preparations; this effect, however, was more evident with the hemin-normal lipopolysaccharide. Further, this enhancing effect of serum was partly reduced in the presence of antibodies raised against the serum lipopolysaccharide-binding protein. The differences in the biological activity of the two lipopolysaccharide preparations could be associated with structural differences detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis. These results indicate that hemin availability affects regulation of an aspect of P. gingivalis virulence, lipopolysaccharide-human neutrophils priming. The reduced capacity for neutrophil priming by hemin-restricted lipopolysaccharide appears to be related to lipopolysaccharide-neutrophil interactions and not to serum factors Targeting bacterial cell-surface components involved in hemin transport might be effective therapy for P. gingivalis-associated periodontal diseases.
authors.affiliation: Department of Periodontology and Oral Biology, Goldman School of Graduate Dentistry, Boston University Medical Center, Massachusetts 02118, USA.
authors.forename: C M
authors.initials: CM
authors.lastname: Champagne
authors.forename: S C
authors.initials: SC
authors.lastname: Holt
authors.forename: T E
authors.initials: TE
authors.lastname: Van Dyke
authors.forename: B J
authors.initials: BJ
authors.lastname: Gordon
authors.forename: L
authors.initials: L
authors.lastname: Shapira
mesh_terms: D016895:Culture Media, Serum-Free; D004305:Dose-Response Relationship, Drug; D006427:Hemin / Q000378:metabolism / Q000502:physiology*; D006801:Humans; D008070:Lipopolysaccharides / Q000494:pharmacology*; D018375:Neutrophil Activation; D009504:Neutrophils / Q000187:drug effects* / Q000378:metabolism; D016966:Porphyromonas gingivalis / Q000378:metabolism* / Q000472:pathogenicity; D013481:Superoxides / Q000378:metabolism; D014774:Virulence
chemical_list: D016895:Culture Media, Serum-Free; D008070:Lipopolysaccharides; D013481:Superoxides; D006427:Hemin