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1、J.gen.Virol.(1988),69,87-98.PrintedinGreatBritain87Keywords:coronavirusJHM/peplomerE2epitopes/neurovirulenceThePeplomerProteinE2ofCoronavirusJHMasaDeterminantofNeurovirulence:DefinitionofCriticalEpitopesbyVariantAnalysisByHELMUTWEGE,*JORNWINTERANDRICHARDMEYER
2、MANNtInstituteofVirologyandImmunobiology,UniversityofWiirzburg,VersbacherStrasse7,D-8700Wiirzburg,F.R.G.(Accepted21September1987)SUMMARYWeselectedmurinecoronavirusJHMvariantsspecificallychangedindefinedantigenicsitesofthepeplomerproteinE2.Variantswereisolated
3、fromthesupernatantsofmonoclonalantibodyhybridomacellcultureswhichcontinuedtosecreteneutralizingantibodiesafterbeinginfectedwithJHM.Comparativeantigenicanalysisandbiologicaltestswereperformedinordertorefineanoperationalepitopemapandtocharacterizefunctionaldoma
4、insimportantforpathogenicity.Thereactionpatterns(neutralization,inhibitionofcellfusion,immunofluorescenceandbindinginELISA)betweenthevariantvirusesandthepanelofmonoclonalantibodieswereverysimilar.Fourgroupsofvariantswerecharacterizedeachofwhichrevealeddistinc
5、tchangesaffectingonedefinedantigenicsite.Theseobservationsindicatedthatatleastfourindependentlymutableantigenicsiteswereassociatedwithdomainsinvolvedincellfusion,neutralizationandpathogenicity(E2-Aa,-Ab,-Baand-Bb).JHMvariantswithalterationsintheE2-Aa,-Abor-Bb
6、sitesweresimilartowild-typevirus.Thesevariantscausedacutehepatitisandencephalomyelitisinmice.Incontrast,JHMvariantswithchangesinsiteE2-Bahadastrongpropensitytoinducechronicdiseaseaccompaniedbydemyelinationpersistingforseveralmonths.INTRODUCTIONPersistentinfec
7、tionofrodentswiththemurinecoronavirusesMHV-JHMorMHV-A59canleadtothedevelopmentofinflammatorydemyelinatinglesionsinbrainandspinalcordsimilartothoseassociatedwithsomehumanneurologicaldisorders.Suchinfectionsthereforeprovideexperimentalmodelstostudypathogeneticm
8、echanisms(SSrensenetal.,1982;Wegeetal.,1982;KnobleretaL,1982;Watanabeetal.,1983;Lavietal.,1984;WeReetal.,1984a;Beushausen&Dales,1985;Massaetal.,1986a,b;Suzumuraetal.,1986).Severalvirusand
