摘要
目的检测PRRSV病毒体外感染原代猪肺泡巨噬细胞(PAM)对猪源IFITM分子mRNA转录水平的影响。方法无菌采集PRRSV、PCV2和JEV阴性仔猪肺脏,分离PAM并用PRRSV病毒接种感染。于感染后不同时间点收集细胞,采用PCR检测PAM细胞PRRSV感染情况,采用荧光定量PCR检测PRRSV在PAM细胞中的复制情况以及IFITM分子mRNA的转录水平变化。结果成功分离到猪原代PAM细胞,接种PRRSV后出现明显的细胞病变。荧光定量PCR检测结果显示,PRRSV感染12 h后可显著上调PAM细胞中IFITM1、IFITM2和IFITM3 mRNA的转录水平,分别在感染后的第36 h、24 h和36 h达到高峰。结论 PRRSV体外感染PAM可显著上调猪IFITM mRNA转录,表明IFITM参与宿主抗PRRSV天然免疫反应,为揭示抗PRRSV天然免疫反应机制提供了理论依据。
Objective To determine the effect of in vitro infection of primary porcine alveolar macrophages(PAM) with the PRRSV virus on the levels of mRNA in porcine IFITM molecules. Methods The lungs of PRRSV, PCV2 and JEV-negative piglets were collected under aseptic conditions, and primary alveolar macrophages were isolated. PAM cells were inoculated with PRRSV, and the infection status at different time points was determined. The replication of PRRSV in PAM cells and the levels of IFITM mRNA were determined using fluorescence quantitative PCR. Results Porcine primary PAM cells were successfully isolated, and a marked cytopathic effect was evident after inoculation with PRRSV. In addition, the levels of IFITM1, IFITM2, and IFITM3 mRNA in PAM cells were significantly up-regulated 12 h after PRRSV infection and peaked at 36 h, 24 h, and 36 h, respectively. Conclusion The levels of IFITM mRNA were significantly up-regulated in PAM cells after in vitro infection with PRRSV, which suggested that IFITMs are involved in the host response to PRRSV. This finding provides a theoretical reference for revealing the mechanism of the innate immune response to PRRSV.
作者
温树波
蒙小刚
霍晓伟
刘锴
宋扬
翟景波
鲁会军
WEN Shu-bo;MENG Xiao-gang;HUO Xiao-wei;LIU Kai;SONG Yang;ZHAI Jin-bo;LU Hui-jun(College of Animal Science and Technology,Inner Mongolia University for Nationalities,Tongliao 028042,Inner Mongolia,China;Brucellosis Prevention and Treatment Engineering Research Center of Inner Mongolia Autonomous Region;Zoonosis Innovation Institute of Inner Mongolia University for Nationalitie;Military Veterinary Institute,Academy of Military Medical Sciences)
出处
《中国病原生物学杂志》
CSCD
北大核心
2021年第8期867-870,877,共5页
Journal of Pathogen Biology
基金
国家重点研发计划项目(No.2018YFD0500803,2018YFD0500104)
内蒙古自治区布鲁氏菌病防治工程技术研究中心开放课题(No.MDK2019082)
内蒙古民族大学博士基金启动项目(No.BS583,BS584)。
关键词
猪繁殖与呼吸综合征
猪肺泡巨噬细胞
荧光定量PCR
干扰素刺激基因
porcine reproductive and respiratory syndrome
porcine alveolar macrophages
real-time fluorescent quantitative PCR
interferon-stimulated genes
