title: Mechanisms of resistance of Porphyromonas gingivalis to killing by serum complement.
abstract: The complement system plays an important role in the host defense against infection, and the formation of the terminal complement complex on the bacterial surface has been shown to be particularly important in killing of gram-negative bacteria. The gram-negative periodontal pathogen Porphyromonas gingivalis is resistant to complement killing, and possible mechanisms suggested for this resistance include protease production and capsule formation. In this study, P. gingivalis Arg- and Lys-gingipain deletion mutants and polysaccharide synthesis deletion mutants have been used to investigate these hypotheses. When Arg- and Lys-gingipain protease mutants were incubated in 20% normal human serum, deposition of complement components on the cell surface was significantly increased compared to that for the wild-type organism. However, despite the increased deposition, the protease mutants maintained resistance to killing and their viability was equal to that seen with heat-inactivated serum. Similar data were obtained when the wild-type organism was treated with gingipain protease inhibitors. K-antigen expression mutants were also resistant to killing. However, mutants which no longer synthesized a surface anionic polysaccharide (APS) (a phosphorylated branched mannan) were extremely sensitive to serum killing. These mutants lack the organized dense glycan surface layer present on the parent strain on the basis of electron microscopy. We conclude that the production of APS at the surface of P. gingivalis rather than Arg- and Lys-gingipain synthesis is the principal mechanism of serum resistance in P. gingivalis.
authors.affiliation: MRC Molecular Pathogenesis Group, Institute of Cell and Molecular Science, Barts and The London, Queen Mary's School of Medicine and Dentistry, 4 Newark Street, London E1 2AT, United Kingdom.
authors.forename: Jennifer M
authors.initials: JM
authors.lastname: Slaney
authors.forename: Alexandra
authors.initials: A
authors.lastname: Gallagher
authors.forename: Joseph
authors.initials: J
authors.lastname: Aduse-Opoku
authors.forename: Keith
authors.initials: K
authors.lastname: Pell
authors.forename: Michael A
authors.initials: MA
authors.lastname: Curtis
mesh_terms: D018829:Adhesins, Bacterial / Q000235:genetics* / Q000378:metabolism; D000942:Antigens, Bacterial / Q000235:genetics; D000954:Antigens, Surface / Q000235:genetics; D015933:Complement C3d / Q000032:analysis / Q000276:immunology; D015938:Complement Membrane Attack Complex / Q000032:analysis / Q000276:immunology*; D003546:Cysteine Endopeptidases / Q000235:genetics* / Q000378:metabolism; D017353:Gene Deletion; D000080867:Gingipain Cysteine Endopeptidases; D006801:Humans; D008351:Mannans / Q000235:genetics / Q000378:metabolism; D008854:Microscopy, Electron; D011135:Polysaccharides, Bacterial / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D016966:Porphyromonas gingivalis / Q000201:enzymology / Q000276:immunology* / Q000648:ultrastructure; D044967:Serum / Q000276:immunology
chemical_list: D018829:Adhesins, Bacterial; D000942:Antigens, Bacterial; D000954:Antigens, Surface; D015938:Complement Membrane Attack Complex; D000080867:Gingipain Cysteine Endopeptidases; C030659:K antigens; D008351:Mannans; D011135:Polysaccharides, Bacterial; D015933:Complement C3d; D003546:Cysteine Endopeptidases