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1、Vaccine,Vol.15,No.2,pp.111-120.1997Copyright01997ElsevierScienceLtd.AllrightsreservedPrintedinGreatMainPII:50264410X(96)0017270264-41oxJ97$17+0.00ELSEVIERAnEscherichiacoliCS3lAfibrillumchimeracapableofinducingmemoryantibodiesinoutbredmicefollowingboosterimmunizationwiththeent
2、ero-pathogeniccoronavirustransmissiblegastroenteritisvirusMauriceDerVartanian*fj,Jean-PierreGirardeau*,ChristineMartin*,ElodieRousset*t,MichelChavarot*,HubertLaude1andMichelContrepois*CS31A$brillaearethin,flexible,heteropolymericproteinaceousappendagesexposedasacapsule-likema
3、terialaroundthecellsurfaceofcertainEscherichiacolistrains.TwoantigenicpeptidesoftheSspikeglycoprotein(TGEV-S)aminoacids(aa)363-371and521-531ofthetransmissiblegastroenteritisvirus(TGEV)weretandemlyintro-ducedintheloop-structured,variableregionaa202-218ofthemajorClpGsubunitprot
4、eincomposingthebulkofCS31A.Theresultinghybridjibrillaewitha25aaheterologouspeptidewereproducedatthecellsurface.Usingamonoclonalantibody(Mab)specificfortheTGEVepitopes,purtjiedhybridjibrillaewereanalysedinWesternblottingundernativeconditions,whichshowedthatthetwoviralepitopesw
5、ererecognizedimmunologicallyasanintegralpartofthehybridfibrillae,andthereforethattheywereantigenicallyactive.Theimmunogenicityofthefusionconstructwasevaluatedwithliverecombinantbacteria,purtjiedhybridClpGmonomers,andpurifiedchimericCS31Apolymers.Whatevertheformofhybridusedasa
6、ntigen,intra-peritoneallyimmunizedoutbredmiceelicitedserumanti-TGEVpeptidesantibodies(Abs)withsign$canttitresandcapableofrecognizingnativeTGEVparticles,indicatingthattheepitopesareexposedinanimmunogenicconformationinallcases.However,virusneutralizationtitreswereonlyobtainedaf
7、terimmunizationwitheitherpur$edpolymersormonomers.Furthermore,4monthsafteranultimateimmunizationwith20,ugofhybridjibrillaemicedevelopedastronganamnesticAbresponseagainstthetwoTGEVpeptidesfollowingboosterinoculationwithvirions.WeconcludethatCS31Ajibrillaecarryingacombinationof
8、TGEVepitopesasinsertcaninduceanimmunologicalmemoryinoutbredanimalsin
