摘要目的 研究重组人干扰素(IFN) α-2b对流感病毒的体外抑制作用.方法 将甲型H1N1流感病毒和乙型流感B/Y病毒接种到体外Vero细胞中,并添加不同浓度的IFNα-2b和奥司他韦共培养.观察并计算病毒空斑数,同时利用荧光定量RT-PCR法比较IFNα-2b和奥司他韦对流感病毒的体外抑制效果,利用荧光显微镜观察病毒核糖核蛋白(RNP)复合物的输出.结果 病毒空斑试验显示,10 μg/μL的IFNα-2b和10 μg/μL奥司他韦对甲型H1N1流感病毒具有明显的抑制作用,病毒空斑数分别为7.5×108和15×108PFU/mL,其中,奥司他韦对病毒的抑制作用较IFNα-2b更强;同样,10 μg/μL IFNα-2b和10μg/μL奥司他韦对乙型流感B/Y病毒均具有明显的抑制作用,病毒空斑形成单位分别为1.1×108和1.5×108PFU/mL.荧光定量RT-PCR检测结果显示,加入10 μmol/LIFNα-2b和10 μmol/L奥司他韦的培养基中甲型H1N1流感病毒和乙型流感B/Y病毒核酸扩增循环阈值(CT)均明显高于流感病毒单独培养组,其中,甲型H1N1流感病毒单独培养组、IFNα-2b和奥司他韦共培养组的病毒核酸CT值分别为16、26和35,乙型流感B/Y病毒单独培养组、IFNα-2b和奥司他韦共培养组的病毒核酸CT值分别为18、27和31.荧光显微镜观察病毒RNP复合物的输出发现,10 μmol/L IFNα-2b能够有效损害感染Vero细胞内病毒RNP复合物的输出.结论 IFNα-2b在体外对甲型H1N1流感病毒和乙型流感B/Y病毒均具有明显的抑制作用.

abstractsObjective To investigate the effect of recombinant human interferon α-2b on influenza virus in vitro.Methods Influenza A virus subtype H1N1 and influenza B/Y virus were inoculated into Vero cells and different concentrations of interferon α-2b and oseltamivir were added.Numbers of virus plaques were observed and calculated,and quantitative RT-PCR were used to assess the inhibitory effect of interferon α-2b and oseltamivir in vitro.The nuclear export of viral ribonucleoprotein (RNP) complexes were monitored under fluorescence microscope.Results Virus plaque test showed that influenza A viruses subtype H1N1 were significantly inhibited when 10 μg/μL interferon α-2b and 10 μg/μL oseltamivir were added,and the numbers of plaques were 7.5 × 108 and 15 × 108 PFU/mL,respectively;the inhibitory effect of oseltamivir was better than that of interferon α-2b.Influenza B/Y viruses were also inhibited when 10 μg/μL interferon α-2b and 10 μg/μL oseltamivir were added,and the numbers of plaques were 1.1 × 108 and 1.5 × 108 PFU/mL,respectively.Quantitative RT-PCR results showed that the cycle threshold (CT) values of influenza A virus subtype H1N1 and influenza B/Y virus were much higher when 10 μmol/L interferon α-2b and 10 μmol/L oseltamivir were added.CT values of influenza A virus subtype H1N1 were 16,26 and 35 before and after inferferon α-2b and oseltamivir were added.CT values of influenza B/Y virus were 18,27 and 31 before and after interferon α-2b and oseltamivir were added.Reduction in the nuclear export of viral RNP in influenza A virus subtype H1N1-infected Vero cells was also observed when 10 μmol/L interferon α-2b were added.Conclusion Interferon α-2b has significantly inhibitory effect on both influenza A virus subtype H1N1 and influenza B/Y virus in vitro.