Cytokine Responses toTreponema pectinovorumandTreponema denticolain Human Gingival Fibroblasts

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Abstract
Human gingival fibroblasts were challenged withTreponema pectinovorumandTreponema denticolato test three specific hypotheses: (i) these treponemes induce different cytokine profiles from the fibroblasts, (ii) differences in cytokine profiles are observed after challenge with live versus killed treponemes, and (iii) differences in cytokine profiles are noted from different gingival fibroblast cell lines when challenged with these treponemes. Three normal gingival fibroblast cell cultures were challenged withT. pectinovorumandT. denticolastrains, and the supernatants were analyzed for cytokine production (i.e., interleukin-1α [IL-1α], IL-1β, IL-6, IL-8, IL-10, gamma interferon, macrophage chemotactic protein 1 [MCP-1], platelet-derived growth factor, tumor necrosis factor alpha, and granulocyte-macrophage colony-stimulating factor). Unstimulated fibroblast cell lines produced IL-6, IL-8, and MCP-1.T. pectinovorumroutinely elicited the greatest production of these cytokines from the fibroblast cell lines, increasing 10- to 50-fold over basal production. WhileT. denticolaalso induced IL-6 and IL-8 production, these levels were generally lower than those elicited by challenge withT. pectinovorum. MCP-1 levels were significantly lower afterT. denticolachallenge, and the kinetics suggested that this microorganism actually inhibited basal production by the fibroblasts. No basal or stimulated production of the other cytokines was observed. Significant differences were noted in the responsiveness of the various cell lines with respect to the two species of treponemes and the individual cytokines produced. Finally, deadT. pectinovorumgenerally induced a twofold-greater level of IL-6 and IL-8 than the live bacteria. These results supported the idea that different species of oral treponemes can elicit proinflammatory cytokine production by gingival cells and that this stimulation did not require live microorganisms. Importantly, a unique difference was noted in the ability ofT. pectinovorumto induce a robust MCP-1 production, whileT. denticolaappeared to inhibit this activity of the fibroblasts. While the general cytokine profiles of the fibroblast cell cultures were similar, significant differences were noted in the quantity of individual cytokines produced, which could relate to individual patient variation in local inflammatory responses in the periodontium.

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