ISG15基因敲除增加小鼠对脑心肌炎病毒(EMCV)的易感性  

Interferon stimulated gene 15knockout mice exhibit higher susceptibility to encephalomyocarditis virus infection

作　　者：何文峰 李琛 李隆熙 董靓靓 杨国庆[1] 常洪涛[2] 刘慧敏[1] HE Wenfeng;LI Chen;LI Longxi;DONG Liangliang;YANG Guoqing;CHANG Hongtao;LIU Huimin(College of Life Science,Henan Agricultural University,Zhengzhou 450002,China;College of Veterinary Medicine,Henan Agricultural University,Zhengzhou 450002,China)

机构地区：[1]河南农业大学生命科学学院,河南郑州450002 [2]河南农业大学动物医学学院,河南郑州450002

出　　处：《中国兽医学报》2023年第9期1873-1880,共8页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(31902268);2021年度河南省高等学校青年骨干教师培养计划资助项目(2021GGJS034)。

摘　　要：本试验旨在通过体内、外试验探究干扰素刺激基因15(interferon-stimulated gene 15,ISG15)的缺失对脑心肌炎病毒(EMCV)易感性的作用。通过ISG15的过表达,分析ISG15对EMCV复制的影响;同时,通过ISG15敲除小鼠进一步验证ISG15对EMCV复制的影响。将ISG15敲除小鼠和野生型小鼠分别分为2组,腹腔注射EMCV后每天记录体质量、临床症状和死亡情况,病毒噬斑检测不同组织的病毒滴度;RT-qPCR和ELISA检测细胞因子IL-1β、IL-6和TNF-α的转录和表达,HE染色分析不同组织的病理变化。结果表明,ISG15过表达后病毒基因转录水平和病毒滴度显著低于对照组细胞;ISG15敲除小鼠与野生型小鼠相比,死亡率显著提高,脑和心脏的病毒滴度也显著升高;血清中细胞因子IL-1β、IL-6和TNF-α的表达量均高于对照组;ISG15-/-小鼠的心肌细胞出现大面积坏死,脑组织出血严重。ISG15敲除显著增强小鼠对EMCV的易感性,并促进小鼠病毒性脑炎发展。这一发现表明,ISG15在体外和体内均抑制EMCV复制,ISG15基因敲除可以提高对EMCV的敏感性,这些发现为EMCV感染提供了新的宿主细胞免疫治疗靶点。To investigate the role of interferon-stimulated gene 15(ISG15)deletion on susceptibility to encephalomyocarditis virus(EMCV),in vitro and in vivo assays were performed in the present study.The roles of ISG15on EMCV infection were analyzed by ISG15overexpression,and further verified using ISG15knockout(ISG15-/-)mice.ISG15-/-mice and wild-type(WT)mice were intraperitoneally injected with EMCV,and their body weight,clinical signs and death were recorded and analyzed.EMCV titers in different tissues were detected by viral plaque,and the transcription and expression of cytokines including IL-1β,IL-6and TNF-αwere detected by RT-qPCR and ELISA.Additionally,histopathological changes of brain and heart tissues were analyzed by H&E staining.The results showed that the viral gene transcript levels and viral titers were significantly lower in cells with ISG15overexpression compared with the control cells.Higher mortality rate and higher viral titers in brain and heart were revealed significantly in ISG15-/-mice compared with WT mice.The expression of cytokines IL-1β,IL-6and TNF-αin serum was higher than those in WT group.Furthermore,massive necrosis of cardiomyocytes and severe hemorrhage in brain tissue were observed in ISG15-/-mice.Taken together,ISG15 knockout significantly enhanced susceptibility to EMCV and promoted the development of viral myocarditis in mice.

关 键 词：ISG15 脑心肌炎病毒 ISG15敲除小鼠 易感性 

分 类 号：Q78[生物学—分子生物学] S852.65[农业科学—基础兽医学]