The Capnocytophaga are inhabitants of the hypoxic human gingival crevice that are normally prevented by neutrophils from causing periodontal and systemic infection. To identify potential nonoxidative bactericidal mechanisms against Capnocytophaga within human neutrophils, gel filtration chromatography was used to fractionate neutrophil granule extracts. Seven granule fractions, designated A through G, were obtained. The Capnocytophaga were most sensitive to killing by fraction D. Fraction D exhibited substantial bactericidal activity under aerobic and anaerobic conditions. The bactericidal activity associated with ion-exchange subfractions D8-D11, which contained primarily cathepsin G as assessed by enzymatic activity, amino acid composition, and NH2-terminal sequence. Heat-inactivation, diisopropylfluorophosphate, PMSF, and N-benzyloxycarbonylglycylleucylphenylalanyl-chloromethyl ketone inhibited bactericidal activity against Capnocytophaga sputigena but not Escherichia coli. We conclude that (a) human neutrophil cathepsin G is an important antimicrobial system against the Capnocytophaga, (b) the bactericidal activity of cathepsin G against Capnocytophaga is oxygen independent, and (c) an intact enzyme active site is involved in the killing of C. sputigena but not E. coli. We suggest that human neutrophil cathepsin G is an important antimicrobial system against certain oral bacteria and that cathepsin G kills bacteria by two distinct mechanisms.
K T Miyasaki, A L Bodeau
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Blood | 1999 |
The acute inflammatory response and the role of phagocytic cells in periodontal health and disease
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Periodontology 2000 | 1997 |
Sensitivity of Periodontal Pathogens to the Bactericidal Activity of Synthetic Protegrins, Antibiotic Peptides Derived from Porcine Leukocytes
KT Miyasaki, R Iofel, RI Lehrer |
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pH-dependent hysteretic behaviour of human myeloblastin (leucocyte proteinase 3)
A Baici, SE Szedlacsek, H Früh, BA Michel |
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Identification of CG-1, a Natural Peptide Antibiotic Derived from Human Neutrophil Cathepsin G
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Neutrophil lysosomal nonoxidative microbicidal proteins in early-onset periodontitis
TF Flemmig, KT Miyasaki |
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Synthetic peptides of human lysosomal cathepsin G with potent antipseudomonal activity
WM Shafer, ME Shepherd, B Boltin, L Wells, J Pohl |
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In vitro Antimicrobial Activity of the Human Neutrophil Cytosolic S-100 Protein Complex, Calprotectin, Against Capnocytophaga sputigena
KT Miyasaki, AL Bodeau, AR Murthy, RI Lehrer |
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Human neutrophil azurocidin synergizes with leukocyte elastase and cathepsin G in the killing of Capnocytophaga sputigena
KT Miyasaki, AL Bodeau |
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Comparison of granule proteins from human polymorphonuclear leukocytes which are bactericidal toward Pseudomonas aeruginosa
KR Wasiluk, KM Skubitz, BH Gray |
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Differential killing of Actinobacillus actinomycetemcomitans and Capnocytophaga spp. by human neutrophil granule components
KT Miyasaki, AL Bodeau, TF Flemmig |
Infection and immunity | 1991 |
In vitro killing of Actinobacillus actinomycetemcomitans and Capnocytophaga spp. by human neutrophil cathepsin G and elastase
KT Miyasaki, AL Bodeau |
Infection and immunity | 1991 |
Human neutrophil peptide defensins induce single strand DNA breaks in target cells
JF Gera, A Lichtenstein |
Cellular Immunology | 1991 |
The Neutrophil: Mechanisms of Controlling Periodontal Bacteria
KT Miyasaki |
Journal of Periodontology | 1991 |