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Characterization of the Cell Surface Substances of Fusobacterium nucleatum that Induce a Chemiluminescence Response by Human Neutrophils Masato Mikami, Chihomi Kato and Kazuko Saito Department of Microbiology, The Nippon Dental University, School of Dentistry at Niigata Accepted for publication 5 July 2003 We previously reported that the surface fraction derived from Fusobacterium nucleatum is responsible for induction of the chemiluminescence (CL) response generated by reactive oxygen intermediates (ROIs) produced by human neutrophils. ROIs are thought to not only kill bacteria but to cause host-cell injury. The purpose of this study was to identify the species of ROIs released by neutrophils stimulated with the bacterial fraction called CL-inducing surface materials (CSM), and to identify the CL-inducing factor in CSM and the molecules involved in signal transduction in neutrophils. The results of inhibition assays using superoxide dismutase and sodium azide demonstrated that superoxide anion produced in neutrophils stimulated with CSM was mainly converted to hypochlorite by the myeloperoxidase-H2O2-Cl- system. A mannose-sensitive or mannan-sensitive lectin-like interaction between CSM and neutrophils was confirmed by sugar inhibition of the CL-stimulating activity of CSM. Studies with specific inhibitors suggested that phosphatidylinositol 3-kinase (PI3K) and phospholipase C (PLC) were involved in the signal transduction in neutrophils in response to CSM stimulation but that pertussis-toxin-sensitive heterotrimeric G protein was not. CSM contains proteins, lipopolysaccharide (LPS), and other polysaccharides, and the 11 kD protein in CSM was found to be essential to the CL-stimulating activity. LPS had an effect that increased the activity of the protein, but neither CD14 nor Toll-like receptor 4 participated in the LPS signaling. A binding assay by the blotting method showed that the 11 kD protein of CSM bound to the 12 kD and 18 kD cell membrane protein of neutrophils. Thus, the results suggested that 11 kD protein and LPS in the surface of F.nucleatum stimulate neutrophils cooperatively and activate PI3K and PLC in neutrophils to produce ROIs. J Jpn Soc Periodontol, 45 : 215∼228, 2003.

Characterization of the Cell Surface Substances of Fusobacterium nucleatum that Induce a Chemiluminescence Response by Human Neutrophils.