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1、©INsTITuTPASTEURIELSEVIERRes.Immunol.Paris19971997,148,247-256Invivostudyofinterferon-alpha-secretingcellsinpigfoetallymphohaematopoieticorgansfollowinginuteroTGEVcoronavirusinjectionI.Splfchal(I)(*),z.ReMkova(I),M.Sinkora(1),J.Sinkora(I),I.Trebichavsky(I),H.Laude(2)andB.Charley(2)(IJDivisionofImmun
2、ologyandGnotobiology,InstituteofMicrobiologyoftheAcademyofSciencesoftheCzechRepublic,54922NovyHnidek(CzechRepublic),and(2)LaboratoiredeVirologieetImmunologiemoleculaires,INRA,78350Jouy-en-Josas(France)SUMMARYNon-infectiousUV·inactivatedtransmissiblegastroenteritisvirus(TGEV)wasprevi•ouslyshowntoindu
3、ceinterferonalpha(IFNa)secretionfollowinginvitroincubationwithbloodmononuclearcells.Inthisstudy,pigfoetusesatdifferentstagesofgestationwereinjectedinuterowith(a)partiallyUV·inactivatedwildTGEVor(b)fUllyUV-inacti•vatedwildordm49-4mutantTGEVcoronavirus.Nucleatedcellsfromfoetalliver,bonemarrow,spleenan
4、dbloodwereisolated10or20hafterinjectionandassayedexvivoforIFNasecretionbyEUSPOTandELISAtechniques.TheadministrationofTGEVinducedIFNa-secretingcellsinfoetalIymphohaematopoieticorgansatmid-gestation.Incontrast,IFNawasnotdetectedincontrolsham-operatedfoetuses.Aspecificpointmutationintheaminoacidsequenc
5、eoftheviralmembraneglycoproteinMofTGEVmutantdm49-4wasassociatedwithlowerorabsentIFNainuteroinducibilitybymutantvirusascomparedwithwildvirus.RowcytometryanalysisdidnotshowdifferencesinleukocytesurfacemarkerexpressionbetweencontrolandTGEV·orbetweendm49-4andwildvirus-treatedfoetuscells,withtheexception
6、ofareductioninpercentagesofpolymorphonuclearcellsinTGEV-treatedIymphohaematopoietictissues,whichisprob•ablyduetoIFNasecretion.ThepresentdataprovidedinvivoevidenceofIFNasecretionatthecelllevelinfoetallymphohaematopoieticorgans.SuchIFNa-secretingcellsinIym•phohaematopoietictissuesmaybethesourceofIFNad
7、etectedduringfoetalinfections.Key-words:Coronavirus,Transmissiblegastroenteritisvirus,IFNa;ELISA,ELISPOT,Foetus,Pig.INTRODUCTIONandanimmunomodulatoraswellasacellgrowthinhibitor.Severalleukocytepopulat
