Interleukin-8 gene expression by human dental pulp fibroblast in cultures stimulated with Prevotella intermedia lipopolysaccharide.
Interleukin (IL)-8 mRNA expression was investigated in human dental pulp fibroblast cultures after stimulation with lipopolysaccharide (LPS) prepared from Prevotella intermedia and inflammatory cytokines. The expression of IL-8 mRNA and the release of IL-8 induced by P. intermedia LPS in pulpal fibroblast cultures were detected by Northern blot analysis and ELISA, respectively. The sufficient concentration of P. intermedia LPS on the IL-8 mRNA expression was 0.1 microgram/ml in pulpal fibroblast cultures. IL-8 mRNA levels began to increase after 2 h of exposure, reached a maximum at 4 to 8 h, and declined after 48 h, reaching the unstimulated level by 60 h. IL-8 production by the pulpal fibroblasts began to increase after 8 h of exposure upon stimulation with 10 microgram/ml of P. intermedia LPS. By contrast Salmonella LPS and synthetic lipid A did not increase IL-8 mRNA concentrations in pulpal fibroblast cultures. Recombinant human IL-1 alpha, beta, and tumor necrosis factor-alpha were capable of stimulating these cells to express IL-8 mRNA but natural human interferon-beta, gamma, and recombinant human IL-6 were incapable in our assay. These results suggest that pulpal fibroblasts are immunoresponsive cells and can elaborate IL-8 upon stimulation with P. intermedia LPS.
Nagaoka
S
S
Department of Operative Dentistry and Endodontology, Kagoshima University Dental School, Japan.
Tokuda
M
M
Sakuta
T
T
Taketoshi
Y
Y
Tamura
M
M
Takada
H
H
Kawagoe
M
M
D000293:Adolescent; D015152:Blotting, Northern; D002478:Cells, Cultured; D016207:Cytokines / Q000494:pharmacology; D003782:Dental Pulp / Q000166:cytology / Q000276:immunology* / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D005347:Fibroblasts / Q000187:drug effects / Q000276:immunology / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D016209:Interleukin-8 / Q000096:biosynthesis*; D008050:Lipid A / Q000494:pharmacology; D008070:Lipopolysaccharides / Q000276:immunology / Q000494:pharmacology*; D008297:Male; D018713:Prevotella intermedia / Q000276:immunology; D012333:RNA, Messenger / Q000032:analysis; D012475:Salmonella / Q000276:immunology
D016207:Cytokines; D016209:Interleukin-8; D008050:Lipid A; D008070:Lipopolysaccharides; D012333:RNA, Messenger