Journal
JOURNAL OF PERIODONTAL RESEARCH
Volume 44, Issue 4, Pages 503-510Publisher
WILEY
DOI: 10.1111/j.1600-0765.2008.01141.x
Keywords
periodontitis; macrophage; Treponema denticola; peptidoglycan; cytokine; kinase
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Funding
- Canadian Institutes of Health Research [MOP-79364]
- Center for Plant and Microbial Complex Carbohydrates [DE-FG09-93ER-20097]
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Background and Objective: Treponema denticola is a key pathogen associated with periodontitis, a chronic inflammatory disease affecting tooth-supporting tissues. In the present study, we investigated the response of human macrophage-like cells to stimulation by peptidoglycan isolated from T. denticola. We also studied the effect of the peptidoglycan preparation on the phosphorylation state of kinases. Material and Methods: Monoblastic leukemia cells (U937 strain) were differentiated into adherent macrophage-like cells using phorbol myristic acid prior to being stimulated for 6 or 24 h with various amounts of T. denticola peptidoglycan. Secreted inflammatory mediators were quantified by enzyme-linked immunosorbent assays. The phosphorylation state of kinases was determined by immunoblotting. Results: The T. denticola peptidoglycan preparation, which was non-toxic for macrophage-like U937 leukemia cells at the concentration used, significantly increased, in a dose-dependent manner, the secretion of the pro-inflammatory cytokines tumor necrosis factor alpha, interleukin-1 beta and interleukin-6. It also increased the secretion of two potent chemokines, interleukin-8 (IL-8) and regulated on activation normal T cell expressed and secreted (RANTES). T. denticola peptidoglycan also induced a significant increase in the secretion of prostaglandin E-2 and matrix metalloproteinase 9 by macrophage-like cells. The phosphorylation state of several kinases, including extracellular regulated protein-serine kinase 2 (+99%), G protein-coupled receptor-serine kinase 2 (+50%), Yes-related protein-tyrosine kinase (+44%) and extracellular regulated protein-serine kinase 1 (+30%) also increased following stimulation with the peptidoglycan preparation. Conclusion: T. denticola peptidoglycan activates intracellular signaling pathways, leading to an increased production of inflammatory mediators by macrophage-like cells.
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