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，10μg／μL的IFNα-2b和10μg／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7．5×10^8和15×10^8PFU／mL，其中，奥司他韦对病毒的抑制作用较IFNα-2b更强；同样，10μg／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1．1×10^8和1．5×10^8PFU／mL。荧光定量RT-PCR检测结果显示，加入10μmol／L IFNα-2b和10μmol／L奥司他韦的培养基中甲型HlN1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，10μmol／LIFNα-2b能够有效损害感染Veto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。
Objective 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 osehamivir 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 osehamivir 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 osehamivir were added, and the numbers of plaques were 7.5 × 10^8 and 15 ×10^8 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 10μg/μL osehamivir were added, and the numbers of plaques were 1.1 × 10^8 and 1.5 × 10^8 PFU/mL, respectively. Quantitative RT-PCR results showed that the cycle threshold （CT） values of influenza A virus subtype H1 N1 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 H1Nl-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.
出处
《中华临床感染病杂志》
CAS
2015年第2期133-138,共6页
Chinese Journal of Clinical Infectious Diseases
基金
国家自然科学基金（81273075）
