血清4型禽腺病毒感染LMH细胞后干扰素刺激基因在转录水平表达量的动态变化

doi:10.16431/j.cnki.1671-7236.2024.05.029

摘要/Abstract

摘要： 【目的】了解鸡肝癌细胞(LMH)感染血清4型禽腺病毒(FAdV-4)后干扰素(IFN)和干扰素刺激基因(ISGs)的表达量变化,为研究FAdV-4与ISGs的相互作用提供参考依据。【方法】试验将FAdV-4感染LMH细胞,观察不同时间点细胞病变,并收集感染0、12、24、36、48、60、72、84和96 h的细胞样品,通过实时荧光定量PCR技术检测感染病毒后不同时间点IFN-α和IFN-β及ISGs基因在转录水平上表达量的动态变化规律。【结果】LMH细胞感染FAdV-4 48 h时出现典型的细胞病变;在感染12 h后,FAdV-4快速增殖,60 h时达到峰值;感染病毒后各时间点,与对照组相比,IFN-α基因转录水平表达量均显著下调(P＜0.05);与对照组相比,在感染后12和24 h,IFN-β基因表达量显著下调(P＜0.05),在36 h时迅速上调表达至峰值(P＜0.05),在48 h时仍维持在较高水平,之后时间点的表达量有所下降且趋于稳定,但仍显著高于对照组(P＜0.05);ISG12、IFIT5及DDIT4基因在感染前期变化不大,在感染后36 h时迅速上调,在感染后96 h达到峰值(P＜0.05);ZFP313、IFITM3及Viperin基因在感染前期表达量变化不大,随后在36 h迅速上调表达至峰值(P＜0.05),在48 h时仍维持在较高水平,之后时间点的表达量有所下降,但仍显著高于对照组(P＜0.05);与对照组相比,CD47、OAS和PKR基因均呈现下调表达。【结论】在FAdV-4感染LMH细胞后,IFN及多种ISGs在转录水平呈现规律性变化,与病毒在LMH细胞中复制存在一定的联系,表明这些天然免疫因子可能在抗FAdV-4反应中发挥着重要作用。

关键词: 血清4型禽腺病毒(FAdV-4); LMH细胞; 干扰素; 干扰素刺激基因; 转录

Abstract: 【Objective】 This study was aimed to investigate the changes of interferon (IFN) and interferon-stimulating genes (ISGs) expression in chicken Leghorn male hepatoma (LMH) infected with Fowl adenovirus serotype-4 (FAdV-4),and provide reference basis for the study of the interaction between FAdV-4 and ISGs.【Method】 The experiment was conducted by infecting LMH cells with FAdV-4,observing the cellular lesions and collecting cell samples at 0,12,24,36,48,60,72,84 and 96 h post-infection,and the dynamic change patterns of the expression of IFN-α and IFN-β and ISGs genes at the transcriptional level at different time points after infection were detected by Real-time quantitative PCR.【Result】 The results showed that after FAdV-4 infection of LMH cells,LMH cells showed typical cytopathic lesions at 48 h.It was detected that FAdV-4 proliferated rapidly at 12 h,and peaked at 60 h. Compared with control group,the expression of IFN-α at the transcriptional level was significantly down-regulated after infection (P＜0.05).Compared with control group,the expression of IFN-β was significantly down-regulated at 12 and 24 h after infection (P＜0.05),and then rapidly and significantly up-regulated to the peak at 36 h (P＜0.05),and remained at a high level at 48 h.After that,the expression declined and stabilized,but was still significantly up-regulated compared with control group (P＜0.05).The expressions of ISG12,IFIT5 and DDIT4 genes did not change much in the early stage of infection,and reached the peak at 96 h after infection (P<0.05).ZFP313,IFITM3 and Viperin genes did not change much in the early stage of infection,and then rapidly and up-regulated to peak at 36 h (P＜0.05),and remained at a high level at 48 h.After that,the expression declined,but was still significantly up-regulated compared with control group(P＜0.05).CD47,OAS and PKR genes showed down-regulated expression compared with control group.【Conclusion】 After FAdV-4 infection of LMH cells,IFN and a variety of ISGs showed regular changes at the transcriptional level,which were associated with the replication of the virus in LMH cells,suggesting that these natural immune factors might play an important role in the anti-FAdV-4 viral response.

Key words: Fowl adenovirus serotype-4 (FAdV-4); LMH cells; interferon; interferon-stimulated genes; transcription

中图分类号:

S852.65

万丽军, 王盛, 谢芝勋, 任红玉, 谢丽基, 范晴, 罗思思, 李孟, 张艳芳, 曾婷婷, 黄娇玲, 张民秀, 谢志勤, 李小凤, 韦悠. 血清4型禽腺病毒感染LMH细胞后干扰素刺激基因在转录水平表达量的动态变化[J]. 中国畜牧兽医, 2024, 51(5): 2081-2090.

WAN Lijun, WANG Sheng, XIE Zhixun, REN Hongyu, XIE Liji, FAN Qing, LUO Sisi, LI Meng, ZHANG Yanfang, ZENG Tingting, HUANG Jiaoling, ZHANG Minxiu, XIE Zhiqin, LI Xiaofeng, WEI You. Dynamic Changes in Expression of Interferon-stimulated Genes at the Transcriptional Level After Infection with Fowl Adenovirus Serotype-4 in LMH cells[J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(5): 2081-2090.

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