禽流感病毒H9N2与新城疫病毒在鸡成纤维细胞中共感染对病毒复制的影响

doi:10.11843/j.issn.0366-6964.2018.11.025

畜牧兽医学报 ›› 2018, Vol. 49 ›› Issue (11): 2521-2528.doi: 10.11843/j.issn.0366-6964.2018.11.025

禽流感病毒H9N2与新城疫病毒在鸡成纤维细胞中共感染对病毒复制的影响

谢军^1,3,4,5, 孙英杰², 周昌娈², 朱善元⁵, 丁铲^2*, 柏家林^1,3,4*

1. 西北民族大学生物工程与技术国家民委重点实验室, 兰州 730030;
2. 中国农业科学院上海兽医研究所, 上海 200241;
3. 西北民族大学, 甘肃省动物细胞工程技术研究中心, 兰州 730030;
4. 西北民族大学生命科学与工程学院, 兰州 730030;
5. 江苏农牧科技职业学院江苏省兽用生物制药高技术研究重点实验室, 泰州 225300

收稿日期:2018-01-23 出版日期:2018-11-23 发布日期:2018-11-23
通讯作者: 柏家林,E-mail:jlbai@xbmu.edu.cn;丁铲,E-mail:shoveldeen@shvri.ac.cn
作者简介:谢军(1990-),男,江苏泰州人,硕士,主要从事动物分子病毒学研究,E-mail:xjjstz2018@163.com
基金资助:
国家自然科学基金重点项目（31530074）；国家自然科学基金面上项目（31372421）；国家自然科学基金青年基金（31400144）

The Effect of Co-infection of H9N2 Subtype Avian Influenza Virus and Newcastle Disease Virus on Virus Replication in the DF-1 of Chicken Embryo Fibroblasts

XIE Jun^1,3,4,5, SUN Ying-jie², ZHOU Chang-luan², ZHU Shan-yuan⁵, DING Chan^2*, BAI Jia-lin^1,3,4*

1. Key Lab of Bioengineering of the State Ethnic Affairs Commission, Northwest Minzu University, Lanzhou 730030, China;
2. Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, China;
3. Gansu Engineering Research Center for Animal Cell, Northwest Minzu University, Lanzhou 730030, China;
4. Faculty of Life Science and Engineering, Northwest Minzu University, Lanzhou 730030, China;
5. Jiangsu Key Laboratory of Veterinary Biotechnologic Drug, Jiangsu Agri-animal Husbandry Vocational College, Taizhou 225300, China

Received:2018-01-23 Online:2018-11-23 Published:2018-11-23

摘要/Abstract

摘要：

临床上禽流感病毒（avian influenza virus，AIV）与新城疫病毒（Newcastle disease virus，NDV）共同感染情况经常发生。为研究AIV与NDV在鸡成纤维细胞DF-1中的共感染对病毒复制的相互影响，首先对NDV和H9N2亚型AIV单独感染DF-1细胞时病毒的复制情况进行了分析，设定以NDV和AIV共感染细胞后12和24 h作为样品采集点，通过Western blot、间接免疫荧光和实时荧光定量PCR检测了两种病毒共感染与单独感染对病毒NP蛋白表达的相互影响。结果显示：Western blot分析表明共感染12和24 h，NDV NP蛋白的表达分别下调了70.6%（P<0.001）和45.7%（P<0.001），AIV NP蛋白的表达分别下调了61.5%（P<0.001）和82.8%（P<0.001）；免疫荧光分析显示共感染组细胞中NDV和AIV的NP蛋白丰度在感染12 h时间点分别下调81.1%（P<0.001）和65.5%（P<0.001），24 h时则分别下调41.2%（P<0.05）和47.4%（P<0.001）；qPCR结果也证实共感染12和24 h组NDV NP的mRNA表达水平分别下调了64.3%（P<0.001）和64.4%（P<0.05），AIV NP的mRNA表达水平分别下调了61.5%（P<0.001）和63.0%（P<0.001）。本研究证明AIV与NDV共感染在细胞水平上可引起病毒复制的相互抑制，为研究NDV和AIV共感染机制在细胞水平上奠定了基础。

Abstract:

The co-infection of avian influenza virus (AIV) and Newcastle disease virus (NDV) in Chickens is common clinical incident. To study the effect of co-infection of AIV (H9N2) and NDV (Herts/33) on virus replication in the DF-1 of chicken embryo fibroblasts, the replication of AIV and NDV in the DF-1 was first analyzed respectively and post-infection 12 and 24 h were selected as sample collecting time. The Western blot, immunofluorescence assay and real-time fluorescence quantitative PCR (qPCR) were conducted to compare the replication of AIV and NDV between co-infection and individual infection groups. Compared with the single virus infection group, the intensity band ratio of NDV NP in Western blot analysis in 12 and 24 h post-infection were down-regulated by 70.6% (P<0.001) and 45.7% (P<0.001),while AIV NP was down-regulated by 61.5% (P<0.001) and 82.8% (P<0.001), respectively. The optical intensity band ratio of AIV NP and NDV NP were decreased by 81.1% (P<0.001) and 65.5% (P<0.001) in 12 h post-infection, 41.2% (P<0.05) and 47.4% (P<0.001) in 24 h post-infection. The relative mRNA expression of NDV NP in co-infection in 12 and 24 h post-infection was descended by 64.3% (P<0.001) and 64.4% (P<0.05), while AIV NP was 61.5% (P<0.001) and 63.0% (P<0.001), respectively. These findings indicate that co-infection of AIV and NDV is able to inhibit their replication at cellular level, laying the foundation of further study the mechanism of common infection of AIV and NDV.

中图分类号:

S852.659.5

谢军, 孙英杰, 周昌娈, 朱善元, 丁铲, 柏家林. 禽流感病毒H9N2与新城疫病毒在鸡成纤维细胞中共感染对病毒复制的影响[J]. 畜牧兽医学报, 2018, 49(11): 2521-2528.

XIE Jun, SUN Ying-jie, ZHOU Chang-luan, ZHU Shan-yuan, DING Chan, BAI Jia-lin. The Effect of Co-infection of H9N2 Subtype Avian Influenza Virus and Newcastle Disease Virus on Virus Replication in the DF-1 of Chicken Embryo Fibroblasts[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(11): 2521-2528.

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禽流感病毒H9N2与新城疫病毒在鸡成纤维细胞中共感染对病毒复制的影响

The Effect of Co-infection of H9N2 Subtype Avian Influenza Virus and Newcastle Disease Virus on Virus Replication in the DF-1 of Chicken Embryo Fibroblasts

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