IBDV和NDV共感染对DF-1细胞的协同致病作用  

Synergistic pathogenic effects of co-infection with IBDV and NDV in DF-1 cells

作　　者：钱晶[1] 欧阳伟[1] 王晶宇[1] 王晓丽[1] 夏兴霞[1] 诸玉梅[1] 王永山[1] QIAN Jing;OUYANG Wei;WANG Jing-yu;WANG Xiao-li;XIA Xing-xia;ZHU Yu-mei;WANG Yong-shan(Institute of Veterinary Medicine,Jiangsu Academy of Agricultural Sciences/Key Laboratory for Veterinary Bio-Product Engineering and Technology,Ministry of Agriculture, Nanjing210014,China)

机构地区：[1]江苏省农业科学院兽医研究所/农业部兽用生物制品工程技术重点实验室

出　　处：《中国兽医学报》2019年第7期1269-1274,共6页Chinese Journal of Veterinary Science

基　　金：国家重点研发计划资助项目(2018YFD0500100,2017YFD0500706,2016YFD0500800);江苏省自然科学基金青年基金资助项目(BK20180299);国家自然科学基金面上资助项目(31772723);江苏省农业科技自主创新资金资助项目(CX(15)1065)

摘　　要：为研究传染性法氏囊病病毒(IBDV)和新城疫病毒(NDV)共感染对DF-1细胞的致病作用,本试验将IBDV和NDV以单独/共同形式感染DF-1细胞,镜下观察可见共感染组最早在感染48h后出现细胞病变现象(CPE),其他感染组在72h后出现明显CPE;细胞活性检测结果显示共感染组在48h出现明显下降,在72h显著低于其他组。感染上清中共感染组与单感染组病毒增殖曲线基本一致,均表现为24~72h处于病毒增殖期。荧光定量PCR检测结果显示,共感染组细胞内NDV核酸载量均显著高于单感染组,其IBDV核酸载量在48h高于单感染组;共感染组MDA5转录水平在24h显著低于其他组,在48h显著高于其他组;各感染组Ⅰ型IFN转录水平受到明显抑制,其中共感染组IFN-β转录水平显著低于单感染组。本研究结果表明IBDV与NDV共感染导致细胞病变加剧并促进胞内病毒复制,明确了病毒共感染具有协同致病作用;还发现病毒共感染导致MDA5-IFN信号通路出现异常,这为下一步解析IBDV与NDV共感染对宿主天然免疫系统的影响及协同致病机制奠定基础。To investigate the pathogenic effects of co-infection with infectious bursal disease virus (IBDV)and Newcastle disease virus(NDV)in DF-1cells,infected with alone or common forms of IBDV and NDV.Under the microscope,the cytopathological effect(CPE)appeared in the co-infection group at 48hours post-infection(hpi),and other groups showed significant CPE at 72hpi.The cell viability results showed that co-infection group decreased at 48hpi,which was significantly lower than other groups at 72hpi.The virus replication curve of co-infection and single-infection groups displayed a similar change,showing the virus proliferation phase at 24-72h.The results of quantitative PCR showed that NDV nucleic acid load in co-infection group was significantly higher than that in single-infection group,and IBDV nucleic acid load was higher than that in the singleinfection group at 48hpi.MDA5transcription level of co-infection group was significantly lower than that of other groups at 24hpi,whereas it was significantly higher than others at 48hpi.Transcription levels of type I IFN in co-infection and single infection groups were inhibited,and IFN-β transcription level in the co-infection group was significantly lower than that in the single infection groups.These results indicated that co-infection of IBDV with NDV lead to increased cytopathic effects and promoted intracellular viral replication,which had synergistic pathogenic effect.Inaddition,co-infection of the virus could lead to abnormalities in the MDA5-IFN signaling pathway, which laid the foundation for the influence of IBDV and NDV co-infection on host natural immune system and the synergistic pathogenic mechanism.

关 键 词：传染性法氏囊病病毒 新城疫病毒 共感染 协同致病 

分 类 号：S855.3[农业科学—临床兽医学]