CA16感染树鼩肺成纤维细胞模型的建立及其受体SCARB2的表达  

Establishment of a tree shrew lung fibroblast model of CA16 infection and the expression of its receptor SCARB2

作　　者：王文广[1] 匡德宣[1] 李娜[1] 陆彩霞[1] 罕园园[1] 仝品芬[1] 孙晓梅[1] 代解杰[1] WANG Wenguang;KUANG Dexuan;LI Na;LU Caixia;HAN Yuanyuan;TONG Pinfen;SUN Xiaomei;DAI Jiejie(Yunnan Innovation Team of Standardization and Application Research in Tree Shrew,Kunming 6500118,China;the Key Laboratory of Yunnan Province for Vaccine Development for Major Infectcious Disease Control)

机构地区：[1]中国医学科学院/北京协和医学院医学生物学研究所树鼩种质资源中心云南省重大传染病疫苗研发重点实验室中国医学科学院医学生物学研究所实验树鼩标准化与应用研究省创新团队

出　　处：《中国实验动物学报》2019年第4期479-484,共6页Acta Laboratorium Animalis Scientia Sinica

基　　金：云南省应用基础研究面上项目(2018FB045);云南省科技人才和平台计划项目(2017HC019);云南省重大科技专项(2017ZF007)~~

摘　　要：目的 探讨建立CA16感染树鼩肺成纤维细胞(tree shrew lung fibroblasts,TSLF)实验模型的可行性。方法 用肠道病毒CA16感染TSLF,以人肺成纤维细胞(KMB-17)作对照,镜下观察细胞病变情况,间接免疫荧光实验观察病毒蛋白和感染相关受体SCARB2蛋白的表达情况,探针法实时荧光定量PCR检测病毒载量,以β-Actin作内参染料法实时荧光定量PCR检测受体SCARB2基因的相对表达量,结合基因序列比对,分析其与感染的相关性。结果 CA16感染TSLF,可引起明显的细胞病变,免疫荧光实验可见病毒和受体SCARB2蛋白,以100TCID50/10^5 cells的比例感染TSLF可在48 h左右病毒载量达到最高,SCARB2基因有与感染相关的高表达,而其基因序列也与人有较高同源性。结论 CA16可感染TSLF,树鼩SCARB2可参与感染。Objective To explore the feasibility of establishing an experimental model of tree shrew lung fibroblasts(TSLF)infected with CA16.Methods TSLF were infected with enterovirus CA16,and human lung fibroblasts(KMB-17)were used as controls.The cytopathic effects(CPE)were observed using an inverted microscope.The expression of viral protein and the SCARB2 receptor protein was observed by indirect immunofluorescence assay,and viral load was detected by TaqMan real-time PCR assay.Usingβ-actin as an internal reference,the relative expression of the SCARB2 gene was detected using SYBR Green real-time PCR,and its correlation with the infection process was analyzed along with the gene sequence alignment.Results CA16 infected TSLF and caused obvious CPE.The virus and SCARB2 protein were observed by immunofluorescence assay.Under the infection rate of 100TCID50/105 cells,the viral load was highest 48 h after infection.The SCARB2 gene had a high expression that was associated with infection,and its gene sequence also has a high homology with that of humans.Conclusion CA16 can infect TSLF and SCARB2 is related to the infection.

关 键 词：CA16 树鼩肺成纤维细胞 感染 SCARB2 

分 类 号：Q95-33[生物学—动物学]