牛病毒性腹泻病毒感染绵羊胃肠道Cajal间质细胞模型的建立  

Establishment of a model of Cajal interstitial cells in the gastrointestinal tract of sheep for the infection of Bovine viral diarrhea virus

作　　者：李胜男 王勇[1] 张辉[1] 冉多良[2] 付强[2] 陈创夫 史慧君[2] LI Shengnan;WANG Yong;ZHANG Hui;RAN Duoliang;FU Qiang;CHEN Chuangfu;SHI Huijun(Key Laboratory of Animal Diseases Control and Prevention,Xinjiang Production and Construction Corps.,Shinhezi 832003,China;College of Veterinary Medicine,Xinjiang Agricultural University,Urumqi 830052,China)

机构地区：[1]新疆生产建设兵团动物疾病防控重点实验室,新疆石河子832003 [2]新疆农业大学动物医学学院,乌鲁木齐830052

出　　处：《黑龙江畜牧兽医》2021年第22期73-75,78,150,共5页Heilongjiang Animal Science And veterinary Medicine

基　　金：新疆生产建设兵团动物疾病防控重点实验室2016年度开放课题(2016BTDJ02;2016BTDJ01)。

摘　　要：为了建立牛病毒性腹泻病毒(BVDV)感染细胞模型,试验将盲传5代的BVDV新疆分离株TC病毒液按照10倍倍比稀释接种至原代绵羊胃肠道Cajal间质细胞(ICC)中,使用Reed-Muench法计算病毒效价,显微镜观察BVDV感染ICC 24 h后细胞形态变化情况,RT-PCR扩增BVDV 5′UTR基因,间接免疫荧光染色检测ICC中的BVDV。结果表明:经计算测得BVDV新疆分离株TC的效价为1×10^(4.6)TCID_(50)/0.1 mL;BVDV感染ICC后出现致细胞病变效应(CPE),细胞出现空泡、萎缩、数量减少及脱落等现象;RT-PCR扩增得到BVDV 5′-UTR基因的目的条带;间接免疫荧光染色可检测到BVDV感染ICC。在ICC中,BVDV盲传5代仍具有较好的感染性和CPE。说明ICC可作为研究BVDV造成胃肠道炎性病变的细胞模型。In order to establish an ideal cell model of Bovine viral diarrhea virus(BVDV)infection,the experiment would blindly pass 5 passages of BVDV Xinjiang isolate TC virus solution which inoculated into primary sheep gastrointestinal interstitial cells of Cajal(ICC)at a 10-fold ratio.Reed-Muench method was used to calculate virus titer;the changes of cell morphology were observed after BVDV infection with ICC for 24 h;RT-PCR amplification of BVDV 5′UTR gene was carried out;indirect immunofluorescence staining was used to detect BVDV in ICC.The results showed that after calculation,the virus titer of BVDV was 1×10^(4.6)TCID_(50)/0.1 mL;after BVDV infection,ICC had a cytopathic effect(CPE),including vacuoles,atrophy,number reduction,and shedding of cells;RT-PCR amplification obtained the target band of BVDV 5′-UTR gene;indirect immunofluorescence staining could detect BVDV infection in ICC.In ICC,blind transmission of BVDV for 5 generations still had good infectivity and CPE.The results suggested that ICC could be used as a cell model for studying gastrointestinal inflammatory lesions caused by BVDV.

关 键 词：牛病毒性腹泻病毒 绵羊 CAJAL间质细胞 细胞模型 细胞病变效应 

分 类 号：Q291[生物学—细胞生物学]