Fibrinogen-neutrophil interactions in response to fMLP and Porphyromonas gingivalis fimbrial peptides.
Porphyromonas gingivalis (P.g) is the primary bacterial agent in many forms of chronic periodontitis. Since polymorphonuclear leukocytes (PMNs) are first-line responders to P.g.- induced inflammation, and fibrinogen is important for in vivo PMN in this disease, we have studied the effect of N-formyl-methionyl-leucyl-phenylalanine (fMLP) (an inflammatory stimulus), P.g. fimbriae and fimbrial peptides (based on FimA, the main structural protein of P.g. fimbriae) on PMN-fibrinogen interactions. Freshly isolated human PMNs were allowed to react with FITC-Fibrinogen and various fimbrial peptides (denoted as FimA followed by amino acid number within whole FimA protein), and FITC-Fibrinogen binding was measured using flow cytometry. Freshly isolated neutrophils were also challenged with Fibrinogen and/or fimbrial peptides to measure IL-8 secretion using ELISA. Our studies show that fibrinogen binding to PMNs is enhanced (p < 0.01) in response to fMLP as well as fimbrial peptides (FimA 61-80) containing the motif LTTE (p < 0.01) in a dose dependent manner but not in response to peptides without that motif. We also observed that fMLP and FimA 61-80 have an additive effect on fibrinogen binding to PMNs (p < 0.05), and fMLP and FimA 171-185 significantly inhibit fMLP-induced fibrinogen binding (p < 0.01). To determine of the role of inflammatory cytokines, we examined IL-8 release from PMNs in response to combinations of P. gingivalis fimbriae, fMLP and fibrinogen. In all cases, IL-8 release increased in a dose-dependent manner (p < 0.05). fMLP-fibrinogen effect on IL-8 release from PMNs was synergistic while fimbriae-fibrinogen effect was additive. In summary, PMN priming by fimbrial peptides facilitates fibrinogen-PMN interaction and may increase inflammation.
Department of Oral Biology, University at Buffalo, Buffalo, New York 14214, USA.
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Sahingur
T K
TK
Boehm
H T
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Sojar
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A
Sharma
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De Nardin
D005340:Fibrinogen / Q000276:immunology; D036781:Fimbriae Proteins / Q000737:chemistry / Q000276:immunology / Q000494:pharmacology*; D010861:Fimbriae, Bacterial / Q000276:immunology; D006801:Humans; D016209:Interleukin-8 / Q000378:metabolism; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology*; D009504:Neutrophils / Q000187:drug effects* / Q000276:immunology; D010455:Peptides / Q000737:chemistry / Q000276:immunology / Q000494:pharmacology*; D016966:Porphyromonas gingivalis / Q000276:immunology*; D011994:Recombinant Proteins / Q000276:immunology
D016209:Interleukin-8; D010455:Peptides; D011994:Recombinant Proteins; C063055:fimbrillin; D036781:Fimbriae Proteins; D009240:N-Formylmethionine Leucyl-Phenylalanine; D005340:Fibrinogen