NDV或H9N2 AIV感染对坦布苏病毒继发感染宿主单核-巨噬细胞的影响  

Effects of Newcastle disease virus(NDV)or avian Influenza virus(H9N2) infection on monocytes/macrophages of Tembusu virus secondary infected hosts

作　　者：张艳慧 马勇[1] 梁雨萌 崔露 陈志杰 李雪峰[1] 胥利 刘柘一 刘胜旺[1] 李海[1] ZHANG Yanhui;MA Yong;LIANG Yumeng;CUI Lu;CHEN Zhijie;LI Xuefeng;XU Li;LIU Zheyi;LIU Shengwang;LI Hai(Division of Avian Infectious Diseases,State Key Laboratory of Veterinary Biotechnology,Harbin Veterinary Research Institute,Chinese Academy of Agricultural Sciences,National Poultry Laboratory Animal Resource Center,Harbin 150069,China)

机构地区：[1]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室/禽呼吸道传染病创新团队/国家禽类实验动物资源库,哈尔滨150069

出　　处：《黑龙江畜牧兽医》2022年第19期15-22,139,共9页Heilongjiang Animal Science And veterinary Medicine

基　　金：国家重点研发计划项目(2016YFD0500107-4);国家蛋鸡产业技术体系项目(CARS-40-K18)。

摘　　要：为研究新城疫病毒(NDV)或H9N2禽流感病毒(AIV)对坦布苏病毒(TMUV)继发感染宿主单核-巨噬细胞的影响及其潜在机制,试验通过实时荧光定量PCR方法检测NDV或H9N2 AIV预先感染鸭单核-巨噬细胞和HD11细胞后再感染TMUV 12,24小时时的TMUV基因组拷贝数,分析是否可用HD11细胞替代鸭单核-巨噬细胞进行后续试验;分别通过高分辨率活细胞共聚焦显微镜和ELISA检测NDV和H9N2 AIV感染12,24 h对HD11细胞中活性氧(ROS)和Ⅰ型干扰素(IFN)含量的影响;通过实时荧光定量PCR方法分别检测PMA、外源Ⅰ型IFN与细胞共孵育后,感染TMUV 12,24小时时ROS和24小时时Ⅰ型IFN对TMUV基因组在细胞中复制的影响。结果表明:感染12,24小时时,在鸭单核-巨噬细胞中,NDV混合感染组的TMUV基因组拷贝数极显著低于TMUV单独感染组(P<0.01);感染12,24小时,H9N2 AIV混合感染组的TMUV基因组拷贝数分别显著和极显著低于TMUV单独感染组(P<0.05,P<0.01)。在HD11细胞中,NDV混合感染组的TMUV基因组拷贝数分别显著和极显著低于TMUV单独感染组(P<0.05,P<0.01);H9N2 AIV混合感染组的TMUV基因组拷贝数极显著低于TMUV单独感染组(P<0.01)。感染12,24小时时,NDV和H9N2 AIV均可极显著提高HD11细胞中ROS的含量(P<0.01)。感染24小时时,NDV可显著提高IFN-α的含量(P<0.05),感染12,24小时时,NDV可极显著提高IFN-β的含量(P<0.01);H9N2 AIV可在感染12,24小时时极显著提高IFN-β的含量(P<0.01)。ROS可在感染TMUV 12,24小时时极显著抑制TMUV基因组的复制(P<0.01),Ⅰ型IFN可在感染TMUV 24小时时显著抑制TMUV基因组的复制(P<0.05)。说明可用HD11细胞替代鸭单核-巨噬细胞进行后续试验;在鸭单核-巨噬细胞中,NDV或H9N2 AIV感染均可以有效抑制TMUV基因组的复制,其作用可能是通过提高ROS和Ⅰ型IFN含量实现的。In order to study the effects of NDV or H9N2 AIV infection on the secondary infection of host monocytes/macrophages by Tembusu virus(TMUV)and its potential mechanism,the TMUV genome copy number in duck monocytes/macrophages and HD11 cells preinfected with NDV or H9N2 AIV and reinfected with TMUV at 12,24 h was detected by real-time fluorescence quantitative PCR,and whether HD11 cells could be used to replace duck monocytes/macrophages for subsequent experiments was analyzed;the effects of NDV and H9N2 AIV infection on the contents of ROS and typeⅠIFN were detected by high-resolution live cell confocal microscopy and ELISA respectively;the effects of ROS and typeⅠIFN on TMUV genome replication in 12,24 h and 24 h after co-incubation of PMA and exogenous typeⅠIFN with cells were de⁃tected by real-time fluorescence quantitative PCR.The results showed that the TMUV genome copy number of the NDV mixed infection group was significantly lower than that of the TMUV single infection group in duck monocytes/macrophages at 12 and 24 h after infection(P<0.01);at 12 and 24 h after infection,the TMUV genome copy number of H9N2 AIV mixed infection group was significantly lower than that of TMUV single infection group(P<0.05,P<0.01).In HD11 cells,the TMUV genome copy number of NDV mixed infection group was significantly lower than that of TMUV alone infection group(P<0.05,P<0.01).The TMUV genome copy number of H9N2 AIV mixed infection group was significantly lower than that of TMUV single infection group(P<0.01).HD11 cells can be used to replace duck monocytes/macrophages for subsequent experiments.NDV and H9N2 AIV could significantly increase the content of ROS in HD11 cells at 12 and 24 h after infection(P<0.01).After 24 h infection,NDV could significantly increase the content of IFN-α(P<0.05),NDV could significantly increase the con⁃tent of IFN-βat 12 and 24 h after infection(P<0.01);H9N2 AIV could significantly increase the content of IFN-βat 12 and 24 h after in⁃fection(P<0.01).ROS could significantly inhibit

关 键 词：坦布苏病毒 新城疫病毒 H9N2禽流感病毒 混合感染 单核-巨噬细胞 

分 类 号：S852.659.6[农业科学—基础兽医学] S852.657[农业科学—兽医学]