EBV LMP1 in Gingival Epithelium Potentially Contributes to Human Chronic Periodontitis via Inducible IL8 Production
EBV LMP1 in Gingival Epithelium Potentially Contributes to Human Chronic Periodontitis via Inducible IL8 Production
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DOI：
10.21873/invivo.11670
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发表时间：
2019-11
期刊：
影响因子：
2.3
通讯作者：
Norihisa Watanabe;Keiko Nodomi;Ryo Koike;A. Kato;O. Takeichi;A. Kotani;T. Kaneko;H. Sakagami
中科院分区：
文献类型：
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作者：
Norihisa Watanabe;Keiko Nodomi;Ryo Koike;A. Kato;O. Takeichi;A. Kotani;T. Kaneko;H. Sakagami
背景/目的：人类慢性牙周炎是一个主要的健康问题。虽然一些口腔细菌已被报道为假定的病原体，但据报道，EB病毒（EBV）与牙周炎的进展有关。然而，EBV在牙周炎病因学中的作用尚不清楚。因此，我们研究了EBV的牙周发病机制，以确认是否EBV编码的潜伏膜蛋白1（LMP 1）诱导人牙龈细胞产生白细胞介素-8（IL-8）。材料与方法：采用实时聚合酶链反应、荧光素酶试验、酶联免疫吸附试验（ELISA）和蛋白质印迹法分别测定Ca 9 -22人牙龈上皮细胞中IL 8 mRNA表达、核因子κ B（NF-κ B B）转录、IL 8产生以及NF-κ B B p65和κ B B α抑制剂（I κ B B α）的磷酸化。使用了两种缺乏负责激活NF-κ B的C-末端激活区（CATR）结构域的LMP 1突变体。结果如下：LMP 1以时间和剂量依赖性方式诱导极高的IL 8产生，其中观察到NF-κB p65和IL-8 B α的同时磷酸化和NF-κ B B的转录。相反，IkB α显性失活突变体显著抑制IL 8的产生和NF-κ B的转录。此外，LMP 1突变体不能诱导IL 8的产生。结论：我们的研究结果表明，由于CATR结构域，LMP 1有助于牙周炎的进展，通过IL 8的生产归因于NF-κ B的激活。
Background/Aim: Human chronic periodontitis is a major health problem. Although some oral bacteria have been reported to be putative pathogens, Epstein–Barr virus (EBV) is reported to be associated with the progression of periodontitis. However, the role of EBV in the aetiology of periodontitis is unknown. Therefore, we investigated periodontal pathogenesis of EBV to confirm whether EBV-encoded latent membrane protein 1 (LMP1) induces Interleukin-8 (IL8) production in human gingival cells. Materials and Methods: Real-time polymerase chain reaction, luciferase assay, enzyme-linked immunosorbent assay (ELISA), and western blotting were performed for determining IL8 mRNA expression, nuclear factor kappa B (NF-ĸB) transcription, IL8 production, and the phosphorylation of NF-ĸB p65 and Inhibitor of kappa B alpha (IĸBα), respectively, in Ca9-22 human gingival epithelial cells. Two LMP1 mutants lacking C-terminal activating region (CATR) domains responsible for activating NF-ĸB were used. Results: Extremely high IL8 production was induced by LMP1 in time- and dose-dependent manner, where simultaneous phosphorylation of NF-κB p65 and IĸBα and transcription of NF-ĸB were observed. On the contrary, IL8 production and NF-ĸB transcription were drastically inhibited by dominant negative mutant of IĸBα. Moreover, the LMP1 mutants failed to induce IL8 production. Conclusion: Our findings suggest that due to CATR domains, LMP1 contributes to the progression of periodontitis via IL8 production attributable to NF-ĸB activation.