Expression and Function of RIG-I in Oral Keratinocytes and Fibroblasts
Expression and Function of RIG-I in Oral Keratinocytes and Fibroblasts
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DOI：
10.1159/000366359
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发表时间：
2014-01-01
影响因子：
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通讯作者：
Kamata, Nobuyuki
中科院分区：
文献类型：
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作者：
Ohta, Kouji;Fukui, Akiko;Kamata, Nobuyuki
背景：口腔粘膜细胞的先天免疫反应可能是宿主抵御病毒感染的第一道防线。视黄酸诱导基因-I (RIG-I) 识别细胞质中的病毒 dsRNA，RIG-I 介导的信号传导调节抗病毒 I 型 IFN 和炎症趋化因子的产生。在这里，我们测试了口腔粘膜细胞通过 RIG-I 参与宿主防御病毒感染的假设。方法：采用RT-PCR和免疫组化方法检测永生化口腔角质形成细胞（RT7）、口腔成纤维细胞（GT1）中RIG-I的表达。 RT7 和 GT1 暴露于 dsRNA 病毒模拟物 Poly I：C-LMW/LyoVec (PLV)。通过实时 RT-PCR 和 ELISA 检查 RIG-I 的 IFN-β 和 CXCL10 表达。通过蛋白质印迹法检测 IRF3 和 STAT1 的磷酸化。结果：RT7和GT1在细胞质中组成型表达RIG-I。此外，PLV 通过 RIG-I 增加 RT7 和 GT1 中 IFN-β 和 CXCL10 的产生，同时 IRF3 和 STAT1 磷酸化。 PLV 诱导的 CXCL10 产生因 IFN-β 的中和和 IFN-α/β 受体 (IFNAR) 的阻断而减弱，表明最初的 IFN-β 通过 RIG-I-IRF3 轴产生，最终通过 IFNAR -STAT1 轴诱导 CXCL10 产生。结论：我们提出，口腔角质形成细胞和成纤维细胞中的 RIG-I 可能逐渐形成针对口腔粘膜病毒感染的宿主防御机制。版权所有 (C) 2014 S. Karger AG，巴塞尔
Background: Innate immune response by oral mucosal cells may be the first line of host defense against viral infection. Retinoic acid-inducible gene-I (RIG-I) recognizes viral dsRNA in the cytoplasm, and RIG-I-mediated signaling regulates antiviral type I IFN, and inflammatory chemokine production. Here, we tested the hypothesis that oral mucosal cell participation in host defense against viral infection via RIG-I. Methods: RIG-I expression was detected in immortalized oral keratinocytes (RT7), oral fibroblasts (GT1) using and RT-PCR and immunohistochemistry. RT7 and GT1 were exposed to dsRNA virus mimic Poly I: C-LMW/LyoVec (PLV). Expression of IFN-beta and CXCL10 via RIG-I was examined by Real-time RT-PCR and ELISA. Phosphorylation of IRF3 and STAT1 were detected by western blotting. Results: RT7 and GT1 constitutively expressed RIG-I in the cytoplasm. Furthermore, PLV increased IFN-beta and CXCL10 productions in both RT7 and GT1 via RIG-I concurrent with phosphorylation of IRF3 and STAT1. PLV-induced CXCL10 production was attenuated by neutralization of IFN-beta and blocking of IFN-alpha/beta receptor (IFNAR), indicating primal IFN-beta production via the RIG-I-IRF3 axis, which eventually induces CXCL10 production via the IFNAR -STAT1 axis. Conclusion: We propose that RIG-I in oral keratinocytes and fibroblasts may cumulatively develop hostdefense mechanisms against viral infection in oral mucosa. Copyright (C) 2014 S. Karger AG, Basel