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1、VirusResearchELSEVlERVirusResearch46(1996)111--124Cooperationbetweentransmissiblegastroenteritiscoronavirus(TGEV)structuralproteinsintheinvitroinductionofvirus-specificantibodiesInrsM.Ant6n1,~,SilviaGonzfileza,MariaJ.Bullidoa,MartaCorsina,CristinaRiscoa,JanP.M.Langeveldb,LuisEnjuanesa,*~CentroNac
2、ionaldeBiotecnologia,CSIC,DepartmentofMolecularandCellBiology,CampusUniversidadAutrnoma,CantoBlanco,28049Madrid,SpainblnstituteforAnimalScienceandHealth(ID-DLO),Postbus65,8200ABLelystad,TheNetherlandsReceived18May1996;accepted5August1996AbstractFollowinginfectionofhaplotypedefinedNIH-miniswinewit
3、hvirulenttransmissiblegastroenteritiscoronavirus(TGEV),isolatedmesentericlymphnodeCD4+T-cellsmountedaspecificproliferativeresponseagainstinfectiousorinactivatedpurifiedvirusinsecondaryinvitrostimulation.Aspecific,dose-dependentresponsetothethreemajorrecombinantviralproteins:spike(S),membrane(M),a
4、ndnucleoprotein(N),purifiedbyaffinitychromatography,wascharacterized.Inductionofinvitroantibodysynthesiswasanalyzed.ThepurifiedrecombinantviralproteinsinducedtheinvitrosynthesisofneutralizingTGEV-specificantibodieswhenporcineTGEV-immunecellswerestimulatedwitheachofthecombinationsmadewithtwoofthem
5、ajorstructuralproteins:S+N,S+M,andtoaminorextentwithM+N,butnotbytheindividualproteins.S-proteinwasdissociatedfrompurifiedvirususingNP-40detergentandthenmicellarS-proteinoligomers(S-rosettes)wereformedbyremovingthedetergent.Theseoccurredpreferentiallybytheassociationofmorethan10S-proteintrimmers.T
6、heseS-rosettesincollaborationwitheitherNorM-proteinselicitedTGEV-specificantibodieswithtitersupto84and60%,respectively,ofthoseinducedbythewholevirus.N-proteincouldbepartiallysubstitutedbya15-merpeptidethatrepresentsaThelperepitopepreviouslyidentified:inN-protein(Ant6netal.(1995)).Theseresultsindi
7、catethattheinductionofhighlevelsofTGEV-specificantibodiesrequiresstimulationbyatleasttwoviralproteins,andthatoptimumresponsesareinducedbyacombinationofS-rosettesandthenucleoprotein.Copyright©1996ElsevierScienceB.V.Keyw
