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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 HCVbutalsocansupportits Longreplicationin vitro.This celllinewithHCV infectionin vitro canserveas a usefultoolforthestudyof themechanismof HCVinfectionandreplication,theevaluation of antiviral agents, andtheprimaryselectionof neutralizationassaysandHCVvaccinedevelopment.