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1、VIROLOGY213,503–516(1995)InductionofAntibodiesProtectingagainstTransmissibleGastroenteritisCoronavirus(TGEV)byRecombinantAdenovirusExpressingTGEVSpikeProteinJUANM.TORRES,*CARLOSSA´NCHEZ,*CARLOSSUN˜E´,*CRISTIANSMERDOU,*,1LUDVIKPREVEC,†FRANKGRAHAM,†andLUISENJUANES**Departmento
2、fMolecularandCellBiology,CentroNacionaldeBiotecnologõ´a,CSIC,CampusUniversidadAuto´noma,Cantoblanco,28049Madrid,Spain;and†DepartmentsofBiologyandPathology,McMasterUniversity,Hamilton,OntarioL8S4K1,CanadaReceivedApril10,1995;acceptedSeptember5,1995Tenrecombinantadenovirusesex
3、pressingeitherfragmentsof1135,1587,or3329ntorthefull-lengthspikegeneoftransmissiblegastroenteritiscoronavirus(TGEV)havebeenconstructed.TheserecombinantsproduceSpolypeptideswithapparentmolecularmassesof68,86,135,and200kDa,respectively.Expressionoftherecombinantantigendrivenby
4、Ad5promoterswasinhibitedbytheinsertionofanexogenousSV-40promoter.Mostoftherecombinantantigensremainintracytoplasmicininfectedcells.Alltherecombinant-directedexpressionproductscontainfunctionalantigenicsitesCandB(Gebaueretal.,1991,Virology183,225–238).Therecombinantantigenof1
5、35kDaandthatof200kDa,whichrepresentsthewholespikeprotein,alsocontainantigenicsitesDandA,whichhavepreviouslybeenshowntobethemajorinducersofTGEV-neutralizingantibodies.Interestingly,hereweshowthatrecombinantSproteinfragmentsexpressingonlysitesCandBalsoinducedTGEV-neutralizinga
6、ntibodies.ThechimericAd5–TGEVrecombinantselicitedlactogenicimmunityinhamsters,includingtheproductionofTGEV-neutralizingantibodies.TheantiserainducedinswinebytheAd5recombinantsexpressingtheamino-terminal26%ofthespikeprotein(containingsitesCandB)orthefull-lengthspikeprotein,wh
7、enmixedwithalethaldoseofviruspriortoadministrationtosusceptiblepiglets,delayedorcompletelypreventedtheinductionofsymptomsofdisease,respectively.q1995AcademicPress,Inc.INTRODUCTIONimmuneresponsetocoronaviruses(Spaanetal.,1990;EnjuanesandVanderZeijst,1995):thespikeprotein(S)Tr
8、ansmissiblegastroenteritiscoronavirus(TGEV)in-(Buchmeieretal.,1984;Cavanagh
