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1、J.Vet.Med.B35,752-759(1988)81988PaulPareyScientificPublishers,BerlinandHamburgISSN0931-1793DepartmentofVeterinaryMicrobiologyandParasitology,SchoolofVeterinaryMedicine,LouisianaStateUniversity,BatonRouge,LA70803,USATheInfluenceoftheHostCellandTrypsinTreatmentonBovineCoronavirusInfectivityK.
2、ST.CYR-COATS,H.R.PAYNEandJ.STORZAddressofauthors:DepartmentofVeterinaryMicrobiologyandParasitology,SchoolofVeterinaryMedicine,LouisianaStateUniversity,BatonRouge,LA70803,USAWith5figures(ReceivedforpublicationJuly17,1988)SummaryWeexaminedtheeffectoftrypsinontheinfectivityofbovinecoronaviruss
3、trainL9(BCV-L9).Setsofculturesofhumanrectaltumor(HRT-18)cells,fourclonesofHRT-18cells,andbovinefetalspleen(DZBFS)cellswereinfectedinduplicatewithserialdilutionsofBCV-L9.Infectedcultureswerepropagatedintrypsin-containingortrypsin-freemedium,andvirusyieldsweremeasuredasplaqueformingunits(PFU)
4、andhemagglutination(HA)titers.TheeffectoftrypsinwasassessedbycomparingPFU:HAratiosobtainedinthepresenceandabsenceoftrypsin.Trypsin-enhancedinfectivitywasevidentinculturesofHRT-18parentcells,D2BFScells,and3ofthe4HRT-18cloneswhenthecultureswereinoculatedwithhighdilutionsofvirus.Thedegreeofenh
5、ancementvariedbetweenthedifferentcellcultures.ThedataindicatethattrypsinmodificationinfluencesBCVinfectivity,butthiseffectdependsonthehostcell.Theeffectofdifferenttrypsin-treatmentconditionsontheproductionofinfectiousviruswasexploredwithDZBFScells.TreatmentofDZBFScellswithtrypsinpriortoinfe
6、ctionhadnoeffectonvirusyield.Theprgsenceoftrypsininculturemediumofinfectedcellsresultedinsignificantlyenhancedvirusyields.Keywords:Bovinecoronavirusinfectivity,trypsinmodificationIntroductionProteolyticcleavageofsurfaceproteinsisaprerequisiteforinfectivityinanumberofvirussystems.Specificall
7、y,orthomyxovirusrequirescleavageofthe75-80KdHAproteinintothe36KdHA1andthe27KdHA2speciesforthevirustobeinfectious.Activationofthecell-fusingpotentialandinfectivityofparamyxovirusisaresultofcleavageoftheFoprecursorproteinintoF1andF2(52and13Kd,respectively)