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.
基金
SupportedbyNationalNaturalScienceFoundationofChina(No.39670672)