禽呼肠孤病毒感染DF1细胞后干扰素刺激基因在转录水平表达量的动态变化

为了解干扰素（interferon,IFN）和干扰素刺激基因在禽呼肠孤病毒（ARV）感染DF1细胞后的表达情况,试验将ARV病毒感染DF1细胞,观察细胞病变,收集感染后0、6、12、24、36、48、72、96 h的细胞样品,抽提反转录成cDNA,通过实时荧光定量PCR技术检测干扰素IFN-α和IFN-β及9种禽源常见干扰素刺激基因在感染后不同时间点在转录水平表达量的动态变化规律。结果显示,在ARV感染DF1细胞后,DF1细胞出现典型的细胞病变,感染后12 h病毒开始快速增殖,在36~96 h维持在较高的水平;IFN-α和IFN-β在转录水平的表达量在感染后均表现为显著下调（P<0.05;P<0.01）;IFI6、OAS、IFIT5、ISG12在转录水平表达量变化规律相似,均呈现显著上调表达（P<0.05;P<0.01）,在感染后96 h达到峰值;其中IFIT5的上调幅度最大,感染后96 h的表达量是0 h的19.62倍（P<0.01）;而Mx、IFITM3、PKR、Viperin、ZAP的表达量变化规律相似,均表现为显著下调表达（P<0.05;P<0.01）,其中Mx、IFITM3、Viperin的下调幅度较大,PKR和ZAP下调幅度很小。说明在ARV感染DF1细胞后,干扰素及多种干扰素刺激基因在转录水平呈现规律性变化,与病毒在DF1细胞中复制存在一定的联系。结果表明,ARV感染后可以诱导多种干扰素刺激基因的表达,这些干扰素刺激基因在抵御ARV病毒的入侵,抑制ARV的复制、释放及病毒的清除中发挥着重要作用。本研究为今后深入研究ARV的致病机理和宿主的抗病毒免疫应答提供了参考。

Altered Gene Expression Profiles of the Interferon Stimulated Genes in DF1 Cells Infected with Avian Reovirus

In order to reveal the expression of interferon(IFN)and interferon stimulated gene in DF1 cells infected with Avian reovirus(ARV),infect DF1 cells with ARV virus,observe cytopathic changes,collect cell samples at 0,6,12,24,36,48,72 and 96 h after infection,and extract reverse transcription into cDNA.Real-time PCR assays was used to detect the dynamic changes of interferon IFN-αand IFN-βand 9 common avian interferon-stimulating genes at the transcription level at different time points after infection.The results showed that after ARV infected DF1 cells,DF1 cells appeared typical cytopathic effect.The virus began to proliferate rapidly at 12 h after infection and maintained at a high level from 36-96 h.The expression of IFN-αand IFN-βat the transcriptional level was significantly down-regulated after infection(P<0.05;P<0.01);IFI6,OAS,IFIT5,and ISG12 had similar changes in the expression level of transcription,and all showed significant up-regulation(P<0.05;P<0.01),reaching peak at 96 h after infection.Among them,IFIT5 had the largest up-regulation,and its expression at 96 h after infection was 19.62 times than that at 0 h(P<0.01).While the expression changes of Mx,IFITM3,PKR,Viperin,and ZAP were similar,and all showed significant down-regulation(P<0.05;P<0.01),among them,the downward adjustment of Mx,IFITM3 and Viperin was larger than that of PKR and ZAP.The results showed that interferon and various interferon-stimulating genes showed regular changes at the transcription level after ARV infection in DF1 cells,which was related to virus replication in DF1 cells.The results showed that ARV infection could induce the expression of variety of interferon-stimulating genes,which played an important role in resisting ARV virus invasion,inhibiting ARV virus replication,release,and virus clearance.This study provided a further research direction of ARV pathogenesis and virus-inhibiting mechanism responses to ARV infection.