牙齦造纖維細胞經不同血清型之怑放線桿菌刺激後產生介白質－6能力之研究

Interleukin-6 Production by Human Gingival Fibroblasts Following Stimulation With Actinobacillus Actinomycetemcomitans

陳慶長(Ching-Charng Chen) ； 張基隆(Kee-Lung Chang) ； 黃正芬(Jeng-Fen Huang) ； 黃景勝(Jiing-Sheng Huang) ； 蔡吉政(Chi-Cheng Tsai)

《The Kaohsiung Journal of Medical Sciences》 14卷6期 (1998/06) Pp. 367-378

https://doi.org/10.6452/KJMS.199806.0367

摘要

Actinobacillus actinomycetemcomitans(Aa), elaborating a multiplicity of virulence factor and tissue-damaging products, is considered an etiological agent in periodontal disease. Serotype b is the most frequently isolated serotype in localized juvenile periodontitis patients, suggesting a particularly high penodontopathic potential for serotype b strains. Interleukin-6(IL-6) plays an important role in the mediation of inflammatory and immune responses as well as in the osteoclastic bone resorption. However, there is little information regarding the effect of the different serotypes of Aa on IL-6 production by human gingival fibroblasts (HGF). Therefore, the purpose of this study was to compare the ability of the three serotypes (a, b, and c) of Aa sonicates to induce the production of IL-6 by HGF. In fibroblast cultures, confluent monolayers of HGF were incubated with sonic extracts of Aa-511 (serotype a), Aa-Y4 (serotpye b), and Aa-652 (serotype c) at various concentrations for 48h at 37 °C in 5% CO2 and air. At the end of the culture period, supernatants were collected and analysed for IL-6 content by using EIA and bioassay. In order to compare the effects of non-lipopolysaccharide (LPS) activation of Aa sonicates on IL-6 production by HGF, we added polymyxin B in cultures with Aa sonicates to bind LPS. The results were summarized as follows. (1) All three serotypes of Aa sonicates had similar dose-dependent stimulant effects on IL-6 production by HGF, and the biological activities of IL-6 correlated with their immunoreactivities. (2) The maximum releases of IL-6 by HGF were achieved at concentrations of 10 to 100 g protein/mL of Aa sonicates, and the ability of Aa-Y4 to induce the release of IL- 6 was higher than that of Aa-51 1 and Aa-652 at these concentrations. (3) Polymyxin B (50 ,&g/ mL) effectively decreased the amounts of IL-6 produced by stimulation of the HGF with 10 ,g protein/mL of Aa sonicates. However, the polymyxin B-treated Aa-Y4 sonicate showed a higher ability to induce the release of IL-6 than the other two strains. These results indicate that Aa-Y4 (serotype b) has a higher potency to induce HGF secretion of IL-6; thus contributing to a comparatively stronger efficacy to the destruction of periodontal tissue in periodonti-tis.

並列摘要

Actinobacillus actinomycetemcomitans(Aa), elaborating a multiplicity of virulence factor and tissue-damaging products, is considered an etiological agent in periodontal disease. Serotype b is the most frequently isolated serotype in localized juvenile periodontitis patients, suggesting a particularly high penodontopathic potential for serotype b strains. Interleukin-6(IL-6) plays an important role in the mediation of inflammatory and immune responses as well as in the osteoclastic bone resorption. However, there is little information regarding the effect of the different serotypes of Aa on IL-6 production by human gingival fibroblasts (HGF). Therefore, the purpose of this study was to compare the ability of the three serotypes (a, b, and c) of Aa sonicates to induce the production of IL-6 by HGF. In fibroblast cultures, confluent monolayers of HGF were incubated with sonic extracts of Aa-511 (serotype a), Aa-Y4 (serotpye b), and Aa-652 (serotype c) at various concentrations for 48h at 37 °C in 5% CO2 and air. At the end of the culture period, supernatants were collected and analysed for IL-6 content by using EIA and bioassay. In order to compare the effects of non-lipopolysaccharide (LPS) activation of Aa sonicates on IL-6 production by HGF, we added polymyxin B in cultures with Aa sonicates to bind LPS. The results were summarized as follows. (1) All three serotypes of Aa sonicates had similar dose-dependent stimulant effects on IL-6 production by HGF, and the biological activities of IL-6 correlated with their immunoreactivities. (2) The maximum releases of IL-6 by HGF were achieved at concentrations of 10 to 100 g protein/mL of Aa sonicates, and the ability of Aa-Y4 to induce the release of IL- 6 was higher than that of Aa-51 1 and Aa-652 at these concentrations. (3) Polymyxin B (50 ,&g/ mL) effectively decreased the amounts of IL-6 produced by stimulation of the HGF with 10 ,g protein/mL of Aa sonicates. However, the polymyxin B-treated Aa-Y4 sonicate showed a higher ability to induce the release of IL-6 than the other two strains. These results indicate that Aa-Y4 (serotype b) has a higher potency to induce HGF secretion of IL-6; thus contributing to a comparatively stronger efficacy to the destruction of periodontal tissue in periodonti-tis.

並列關鍵字

A. actinomycetemcomitans ； firbroblasts ； interleukin-6