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Effects of IL-17 on Human Gingival Fibroblasts

¹ Department of Periodontology,
² Research Unit for Periodontal Disease,
³ Immunology Laboratory, and
⁴ Department of Oral Maxillofacial Surgery, Faculty of Dentistry, Chulalongkorn University, Henri Dunant Rd., Bangkok 10330, Thailand; and
⁵ Department of Immunology and Medicine, US Army Medical Component, Armed Forces Research Institute of Medical Sciences (AFRIMS), Bangkok, Thailand

View larger version (33K): [in this window] [in a new window]   Figure 1. Flow cytometric analysis of HLA-DR, CD40, and ICAM-1 expression by HGFs after single stimulation with IL-17 or IFN-. HGF cultures were stimulated with indicated concentrations of IL-17 (A) and IFN- (B) for 5 days. Culture medium was used as a control. The surface molecule expression was determined by flow cytometry. Dotted lines are isotype controls, shaded areas are unstimulated HGFs, and solid lines represent cytokine stimulation. The x-axis and the y-axis indicate the relative fluorescence intensity and cell number, respectively, and the number appearing in the upper right corner of each histogram indicates mean fluorescence intensity (MFI). Data are representative of 13 separate experiments.

View larger version (32K): [in this window] [in a new window]   Figure 2. Flow cytometric analysis of HLA-DR, CD40, and ICAM-1 expression by HGFs after combined stimulation with IL-17 and IFN-. HGF cultures were stimulated with indicated concentrations of IL-17 (A), IFN- (B), or combined cytokines (C) for 5 days. Culture medium was used as a control. The surface molecule expression was determined by flow cytometry. Dotted lines are isotype controls, shaded areas are unstimulated HGFs, and solid lines represent cytokine stimulation. The x-axis and the y-axis indicate the relative fluorescence intensity and cell number, respectively, and the number appearing in the upper right corner of each histogram indicates mean fluorescence intensity (MFI). Data are representative of 13 separate experiments.

View larger version (17K): [in this window] [in a new window]   Figure 3. IL-8 production by HGFs after single stimulation with IL-17, IFN-, and combined cytokine stimulation. HGF cultures were stimulated with indicated concentrations of IL-17 (A), IFN- (B), and the combined cytokines (C) for 2 days. Culture medium was used as a control. The culture supernatants were collected, and IL-8 concentrations were determined by ELISA. Data in (A) and (B) are mean ± SEM of 15 separate experiments. Each symbol in (C) represents IL-8 production of each individual culture. Horizontal lines are means of 15 separate experiments (*P < 0.05, compared with the sum of two individual cytokine stimulations).

View larger version (21K): [in this window] [in a new window]   Figure 4. Expression of IDO in HGFs after stimulation with IL-17, IFN-, or combined cytokines. HGF cultures were stimulated with indicated concentrations of IL-17, IFN-, and combined cytokines. Culture medium was used as a control. Stimulated HGFs were harvested, and mRNA expression of IDO was analyzed by RT-PCR. GAPDH mRNA was used as an internal control. Data are representative of 4 separate experiments (A). Mean relative expression of IDO:GAPDH ± SEM from 4 separate experiments is shown (*P < 0.05, compared with the sum of IL-17 and IFN- [1 U/mL] stimulation; **P < 0.05, compared with the sum of IL-17 and IFN- [5 U/mL] stimulation; ***P < 0.05, compared with the sum of IL-17 and IFN- [25 U/mL] stimulation) (B). To assess IDO activity, we determined the concentrations of kynurenine in culture supernatants. Data shown are mean concentrations of kynurenine ± SEM from 4 separate experiments (*P < 0.05, compared with the sum of IL-17 and IFN- [1 U/mL] stimulation; **P < 0.05, compared with the sum of IL-17 and IFN- [5 U/mL] stimulation; ***P < 0.05, compared with the sum of IL-17 and IFN- [25 U/mL] stimulation) (C).

Journal of Dental Research, Vol. 87, No. 3, 267-272 (2008)
DOI: 10.1177/154405910808700314


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