Calprotectin release from human neutrophils is induced by Porphyromonas gingivalis lipopolysaccharide via the CD‐14–Toll‐like receptor–nuclear factor κB pathway

Publisher Website
Google Scholar
Abstract
Objectives: Calprotectin is a cytosolic protein with antibacterial action in leukocytes and its level increases in some inflammatory diseases, including periodontal diseases, rheumatoid arthritis and ulcerative colitis. Recently, we found that the lipopolysaccharide of Porphyromonas gingivalis (P‐LPS) induced calprotectin release from human neutrophils. P‐LPS, a major virulence factor of periodontal pathogens, is known to induce the production and release of inflammatory cytokines through CD14, Toll‐like receptor (TLR) and nuclear factor κB (NF‐κB). In the present study, we investigated whether calprotectin release by P‐LPS is induced via the CD14–TLR–NF‐κB pathway and the cellular mechanism of calprotectin release in human neutrophils. Material and methods: Human neutrophils were isolated from the peripheral blood of healthy donors and pre‐incubated in medium containing antibodies against CD14, TLR2 and TLR4, or several inhibitors of NF‐κB, microtubules and microfilaments, and then incubated with P‐LPS. The calprotectin amount in the culture medium was determined using ELISA, and the nuclear extracts from cells were used for the examination of NF‐κB binding activity using electrophoretic mobility shift assays. Results: P‐LPS increased calprotectin release from neutrophils and its induction was inhibited by anti‐CD14 and anti‐TLR2 antibodies, but not by two anti‐TLR4 antibodies. NF‐κB inhibitors suppressed P‐LPS‐induced NF‐κB binding activity and calprotectin release. The inhibitors of microtubule and microfilament polymerization significantly decreased P‐LPS‐induced calprotectin release. Conclusion: These results suggest that calprotectin release is induced by P‐LPS via the CD14–TLR2–NF‐κB signal pathway in human neutrophils and may be dependent on microtubule and microfilament systems.

This publication has 36 references indexed in Scilit: