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1、VIROLOGY233,327–338(1997)ARTICLENO.VY978598ExpressionofInterferon-gbyaCoronavirusDefective-InterferingRNAVectorandItsEffectonViralReplication,Spread,andPathogenicity,1,,,,,XumingZhang,*DavidR.Hinton,*†DanielJ.Cua,*‡StephenA.Stohlman,*‡andMichaelM.C.Lai*‡§*DepartmentofNeurology,†Departmentof
2、Pathology,‡DepartmentofMolecularMicrobiologyandImmunology,and§HowardHughesMedicalInstitute,UniversityofSouthernCaliforniaSchoolofMedicine,LosAngeles,California90033-1054ReceivedNovember13,1996;returnedtoauthorforrevisionMarch12,1997;acceptedMay5,1997Adefective-interfering(DI)RNAofthemurinec
3、oronavirusmousehepatitisvirus(MHV)wasdevelopedasavectorforexpressinginterferon-g(IFN-g).ThemurineIFN-ggenewasclonedintotheDIvectorunderthecontrolofanMHVtranscriptionalpromoterandtransfectedintoMHV-infectedcells.IFN-gwassecretedintoculturemediumasearlyas6hrposttransfectionandreachedapeakleve
4、l(upto180U/ml)at12hrposttransfection.TheDI-expressedIFN-g(DE-IFN-g)exhibitedanantiviralactivitycomparabletothatofrecombinantIFN-gandwasblockedbyaneutralizingmonoclonalantibodyagainstIFN-g.TreatmentofmacrophageswithDE-IFN-gselectivelyinducedtheexpressionofthecellularinduciblenitricoxidesynth
5、aseandtheIFN-g-inducingfactor(IGIF)butdidnotaffecttheamountsoftheMHVreceptormRNA.AntiviralactivitywasdetectedonlywhencellswerepretreatedwithIFN-gfor24hrpriortoinfection;noinhibitionofvirusreplicationwasdetectedwhencellsweretreatedwithIFN-gduringorafterinfection.Furthermore,additionofIFN-gto
6、getherwithMHVdidnotpreventinfection,butappearedtopreventsubsequentviralspread.MHVvariantswithdifferentdegreesofneurovirulenceinmicehadcorrespondinglydifferentlevelsofsensitivitiestoIFN-gtreatmentinvitro,withthemostvirulentstrainbeingmostresistanttoIFN-gtreatment.Infectionofsusceptiblemicewi
7、thDE-IFN-g-containingviruscausedsignificantlymilderdisease,accompaniedbymorepronouncedmononuclearcellinfiltratesintotheCNSandlessvirusreplication,thanthatcausedbyviruscontainingacontrolDIvector.ThisstudythusdemonstratesthefeasibilityandusefulnessofthisMH
