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1、0INSTITUTPASTEURIELSEVIERRexImmunol.Paris19931993,144,111-120Enrichmentofcoronavirus-inducedinterferon-producingbloodleukocytesincreasestheinterferonyieldpercell:astudywithpigleukocytesW.NowackiandB.Charley(*)LaboratoiredeVirologieetImmunologicmohkula
2、ires,INRA,78350Jouy-en-Josas(France)SUMMARYPorcineperipheralbloodmononuclearcells,whichsecreteIFNuinresponsetoacoronavirus,transmissiblegastroenteritisvirus,weredetectedbyafilterimmunoplaqueassay(ELISPOT).IFNa-producingcells(IPC),whicharepresentatalow
3、frequencyintheblood,couldbeenrichedupto1OO-foldbysequentialdepletionofplastic-adherentcellsandcellfractionationonmetrizamidedensitygradients.IPCwerepresentinthenon-adherentlow-densitycellsubpopulation.Cellselectionexperimentsusingantibody(Ab)-coatedim
4、munomagneticbeadsrevealedthatporcineIPCcouldbepositivelyselectedbyanti-CD4or-SLA-class-11Ab,butnotbyanti-CD2or-CD8Ab.TheestimatedIFNyieldperIPCwasfoundtoincreasewhenIPCwereassayedathigherconcentrations.ThesedatasuggestthatIPCrepresentauniqueanddistinc
5、tcellpopulationintheblood,whichcouldsecretehigheramountsofIFNfollowingitsaccumulationatasiteofviralinfection.Key-words:Coronavirus,Leukocyte,Interferonalpha,Transmissiblegastroenteritisvirus;IPC,ELISPOT,Immunobeads,PBMC,mAb.INTRODUCTIONviralreplicatio
6、ninIFNa-producingleukocytes,butrather,maybetheconsequenceofmem-Peripheralbloodmononuclearcells(PBMC)braneinteractionsbetweenIFNaproducercellsfromseveralmammalianspeciesarecapableofandviralproteinspresentatthesurfaceofvi-secretinginterferon-a(IFNa)invi
7、trofollowingrusparticlesorvirus-infectedcells.Indeed,sever-briefexposuretonon-infectiousviralstructures,alreportsindicatethatglycosylatedviralproteinssuchasinactivatedvirusparticlesormaybetheessentialtriggersforIFNainductionglutaraldehyde-fixedvirus-i
8、nfectedcellmonolay-(Itoetal.,1978;Lebon,1985;Charleyetal.,ers(Lebonetal.,1982;Capobianchietal.,1985;1991;Laudeetal.,1992;Capobianchietal.,Kuraneetal.,1986).Thesefindingssuggestthat1992).CellswhichproduceIFNainresponsetothisIFNa-inductionmechan
