CA16 感染树鼩肺成纤维细胞模型的建立及其受体SCARB2 的表达
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(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,昆明 650118)

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Q95-33

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Establishment of a tree shrew lung fibroblast model of CA16 infection and the expression of its receptor SCARB2
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(Center of Tree Shrew Germplasm Resources, Institute of Medical Biology, the Chinese Academy of Medical Sciences andPeking Union Medical College; the Key Laboratory of Yunnan Province for Vaccine Development for Major InfectciousDisease Control; Yunnan Innovation Team of Standardization and Application Research in Tree Shrew, Kunming 6500118, China)

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    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 /105 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。

    Abstract:

    Objective To explore the feasibility of establishing an experimental model of tree shrew lungfibroblasts (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. Theexpression of viral protein and the SCARB2 receptor protein was observed by indirect immunofluorescence assay, and viralload was detected by TaqMan real-time PCR assay. Using β-actin as an internal reference, the relative expression of theSCARB2 gene was detected using SYBR Green real-time PCR, and its correlation with the infection process was analyzedalong with the gene sequence alignment. Results CA16 infected TSLF and caused obvious CPE. The virus and SCARB2protein were observed by immunofluorescence assay. Under the infection rate of 100TCID50 /105 cells, the viral load washighest 48 h after infection. The SCARB2 gene had a high expression that was associated with infection, and its genesequence also has a high homology with that of humans. Conclusion CA16 can infect TSLF and SCARB2 is related to the infection.

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王文广,匡德宣,李娜,陆彩霞,罕园园,仝品芬,孙晓梅,代解杰. CA16 感染树鼩肺成纤维细胞模型的建立及其受体SCARB2 的表达[J].中国实验动物学报,2019,27(4):479~484.

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  • 收稿日期:2019-03-28
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  • 在线发布日期: 2019-09-04
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