Characteristics of HCV replication and expression in a cultured human liver carcinoma cell line in vitro

Characteristics of HCV replication and expression in a cultured human liver carcinoma cell line in vitro

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摘要 Objective:To establisha cellculturesystemto supportHCVlong-termreplicationin vitro.Methods:A human hepatomacellline7721was testedfor itssusceptibilityto HCVby incubatingwitha serumfromchronichepatitisC pa-tient.Cellsandsupernatantof theculturemediumwereharvestedat varioustime-phasesduringtheculturingperiods.The presenceof HCVRNA,theexpressionof HCVantigensincellsand/orsupernatantwereexaminedwithRT-PCR,in situ hybridizationandimmunohistochemistryrespectively.Results:It was foundthattheintracellularHCVRNAwas firstde-tectedon the2nddayafterculture,andthencouldbe intermittentlydetectedinbothcellsandsupernatantovera periodof atleast3monthsafterculture.HCVNS3,CP10antigenswereexpressedinthecells.Thefreshcellscouldbe infected withthesupernatantfromculturedinfectedcellsandthetransmissionof viralgenomefromHCV-infected7721cellsto pe-ripheralbloodmononuclearcells(PBMCs)wasalsoobserved.Conclusion:Ourfindingssuggestthatthehumanlivercarci-nomacellline7721isnotonlysusceptibleto HCVbutalsocansupportitslongreplicationin vitro.ThiscelllinewithHCV infectionin vitro canserveas a usefultoolforthestudyof themechanismof HCVinfectionandreplication,theevaluation of antiviralagents,andtheprimaryselectionof neutralizationassaysandHCVvaccinedevelopment. Objective:To establisha cellculturesystemto supportHCVlong-termreplicationin vitro.Methods:A human hepatomacellline7721was testedfor itssusceptibilityto HCVby incubatingwitha serumfromchronichepatitisC pa-tient.Cellsandsupernatantof theculturemediumwereharvestedat varioustime-phasesduringtheculturingperiods.The presenceof HCVRNA,theexpressionof HCVantigensincellsand/orsupernatantwereexaminedwithRT-PCR,in situ hybridizationandimmunohistochemistryrespectively.Results:It was foundthattheintracellularHCVRNAwas firstde-tectedon the2nddayafterculture,andthencouldbe intermittentlydetectedinbothcellsandsupernatantovera periodof atleast3monthsafterculture.HCVNS3,CP10antigenswereexpressedinthecells.Thefreshcellscouldbe infected withthesupernatantfromculturedinfectedcellsandthetransmissionof viralgenomefromHCV-infected7721cellsto pe-ripheralbloodmononuclearcells(PBMCs)wasalsoobserved.Conclusion:Ourfindingssuggestthatthehumanlivercarci-nomacellline7721isnotonlysusceptibleto HCVbutalsocansupportitslongreplicationin vitro.ThiscelllinewithHCV infectionin vitro canserveas a usefultoolforthestudyof themechanismof HCVinfectionandreplication,theevaluation of antiviralagents,andtheprimaryselectionof neutralizationassaysandHCVvaccinedevelopment.

作者宋志强郝飞闵峰刘道践

出处《Journal of Medical Colleges of PLA(China)》 CAS 2001年第3期214-219,共6页 中国人民解放军军医大学学报（英文版）

基金 SupportedbyNationalNaturalScienceFoundationofChina(No.39670672)

关键词 HEPATITIS C VIRUS CELL culture CELL model REPLICATION hepatitisC virus cellculture cellmodel replication

分类号 R730.2 [医药卫生—肿瘤]

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Journal of Medical Colleges of PLA(China)

2001年第3期

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Characteristics of HCV replication and expression in a cultured human liver carcinoma cell line in vitro

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