Regulation of interleukin-6 expression in human dental pulp cell cultures stimulated with Prevotella intermedia lipopolysaccharide.
Interleukin (IL)-6 expression in human dental pulp cell cultures after stimulation with Prevotella intermedia lipopolysaccharide (LPS) was investigated by Northern blot analysis, enzyme immunoassay, and bioassay. The IL-6 mRNA expression began to increase after 1 hr and continued after up to 8 hr of exposure on stimulation with 10 microg/ml of P. intermedia LPS. The bioactivity was dose-dependent on the concentration of P. intermedia LPS (0 to 100 microg/ml). The IL-6 mRNA expression was inhibited by actinomysin D and super-induced by cycloheximide. Anti-CD14 monoclonal antibody (MY4) inhibited the IL-6 mRNA expression when administered at a 0.5 microg/ml concentration before stimulation with P. intermedia LPS at 1 microg/ml. The immunoregulatory cytokines (interferon-gamma, IL-10, and IL-4) inhibited LPS-induced IL-6 production with a combined treatment. These results suggest the IL-6 expression by pulp cell cultures is CD14-dependent and regulated at the transcriptional level, and a combined treatment with immunoregulatory cytokines may be effective for control of pulpal inflammation due to P. intermedia LPS.
Tokuda
M
M
Department of Operative Dentistry and Endodontology, Kagoshima University Dental School, Japan.
Sakuta
T
T
Fushuku
A
A
Torii
M
M
Nagaoka
S
S
D000911:Antibodies, Monoclonal / Q000276:immunology; D001681:Biological Assay; D015152:Blotting, Northern; D018929:Cell Culture Techniques; D003513:Cycloheximide / Q000494:pharmacology; D003609:Dactinomycin / Q000494:pharmacology; D003782:Dental Pulp / Q000166:cytology / Q000187:drug effects / Q000276:immunology* / Q000382:microbiology; D005786:Gene Expression Regulation; D006801:Humans; D007124:Immunoenzyme Techniques; D007371:Interferon-gamma / Q000494:pharmacology; D016753:Interleukin-10 / Q000494:pharmacology; D015847:Interleukin-4 / Q000494:pharmacology; D015850:Interleukin-6 / Q000096:biosynthesis* / Q000235:genetics; D018950:Lipopolysaccharide Receptors / Q000276:immunology; D008070:Lipopolysaccharides / Q000276:immunology*; D018713:Prevotella intermedia / Q000276:immunology*; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D011671:Pulpitis / Q000276:immunology / Q000382:microbiology; D012333:RNA, Messenger / Q000235:genetics; D013997:Time Factors; D014158:Transcription, Genetic / Q000235:genetics
D000911:Antibodies, Monoclonal; D015850:Interleukin-6; D018950:Lipopolysaccharide Receptors; D008070:Lipopolysaccharides; D011500:Protein Synthesis Inhibitors; D012333:RNA, Messenger; D016753:Interleukin-10; D003609:Dactinomycin; D015847:Interleukin-4; D007371:Interferon-gamma; D003513:Cycloheximide