蓝舌病毒通过诱导巨噬细胞产生干扰素抑制HBV增殖

Bluetongue virus inhibits HBV proliferation by inducing macrophages to produce interferon

目的探讨蓝舌病毒(bluetongue virus,BTV)16激活巨噬细胞天然免疫的机制及BTV16刺激后的巨噬细胞培养上清对HBV的抑制作用.方法采用Real-time PCR分别检测BTV未预处理或预处理12 h后巨噬细胞中GAPDH、IFN-α、IFN-β、IRF-3、IRF-5和IRF-7的mRNA表达水平,采用Western blot分别检测IRF3、IRF5、IRF7蛋白的表达.使用ELISA法检测巨噬细胞培养基上清中HBsAg及HBeAg的表达.结果 BTV16能够激活TLR3/干扰素信号转导通路,诱导巨噬细胞高表达Ⅰ型干扰素并诱导干扰素调节因子高表达,20%体积比的BTV刺激后的巨噬细胞培养上清对HBV复制即有较强的抑制作用(P<0.05),Ⅰ型干扰素受体抗体预处理后,BTV16刺激巨噬细胞的上清对HBV释放的抑制作用显著减弱(t=14.031,P=0.003).结论 BTV16能够激活巨噬细胞的天然免疫反应,显著抑制HBV在肝细胞的复制,为临床治疗乙型肝炎提供了新思路.

Objective To investigate the mechanism of bluetongue virus (BTV) 16 activating macrophage innate immunity and the inhibitory effect of macrophage culture supernatant stimulated by BTV16 on HBV. Methods The expression levels of GAPDH, IFN-α, IFN-β, IRF-3, IRF-5 and IRF-7 mRNA in macrophages which is pretreated by BTV for 12 hours or not were detected by Real-time PCR, respectively. The expression of IRF3, IRF5 and IRF7 protein were detected by Western blot. The expression of HBsAg and HBeAg in macrophage culture medium were detected by ELISA. Results BTV16 could activate TLR3/IFN signaling pathway and induce macrophages to overexpress interferon Ⅰ, and sufficient to induce high expression of interferon regulatory factor. Twenty percent volume ratio of macrophage culture medium stimulated BTV had a strong inhibitory effect on HBV replication (P < 0.05). After pretreatment of interferon Ⅰ receptor antibodies, the inhibitory effect of macrophage culture medium stimulated BTV on HBV releasing significantly decreased (t = 14.031, P = 0.003). Conclusion BTV16 can activate the innate immune response of macrophages and significantly inhibit the replication of HBV in hepatocytes, which provides a new idea for clinical treatment of hepatitis B.