冠状病毒Coronavirusl论文-1990 Initial events in bovine coronavirus infection_ analysis through immunogold probes and lysosomotropic inhibitors.pdf

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1、ArchVirol(1990)114:175-189VirologyArchives©bySpringer-Verlag1990Initialeventsinbovinecoronavirusinfection:analysisthroughimmunogoldprobesandlysosomotropieinhibitorsH.R.Paynel,J.Storz1,andW.G.Henk2Departmentsof1VeterinaryMicrobiologyandParasitologyandof2Vete

2、rinaryAnatomyandFineStructure,LouisianaStateUniversity,BatonRouge,Louisiana,U.S.A.AcceptedMay14,1990Summary.Theearlyeventsintheinfectionofhumanrectaltumorcellsbybovinecoronaviruswereinvestigatedbycolloidalgold-mediatedimmunoelec-tronmicroscopyandbyanalysiso

3、ftheeffectoflysosomotropicweakbasesonvirusyield.Electronmicroscopicstudiesrevealedsitesoffusionbetweenthevirusenvelopeandtheplasmalemmabutfusioneventsalongintracellularmem-braneswerenotobserveddespiteextensivesearches.Virion-antibody-colloidalgoldcomplexesw

4、ere,infact,endocytosedbysynchronouslyinfectedcells.Thesecomplexeswereapparentlynon-infectious,andtheyaccumulatedinvacuolesthatresembledsecondarylysosomes.Exposureofcellstoammoniumchlorideortomethylamineduringthefirsthourofinfectionhadlittleinhibitoryeffecto

5、ntheproductionofinfectiousvirus.Chloroquinetreatmentswereinhibitorybutthiseffectdependedonrelativelylateeventsintheinfectiousprocess.ThechloroquineinhibitorystepblockedinfectionofvirusadsorbedtocellsthatwereexposedtobuffersinthepHrangeof4.4to8.4.Thesefindin

6、gsindicatethatBCVpenetratesitshostcellbydirectfusionwiththeplasmalemmaanddoesnotrequireanacidicintracellularcompartmentforinfectiousentry.IntroductionBovinecoronavirus(BCV)isamemberofthefamilyCoronaviridaeofenvelopedRNAviruses1-31].TheBCVisolatesstudiedinso

7、medetailagglutinateeryth-rocytesofrodentsandthusbelongtothehemagglutinatingcoronaviruses.Uniquely,theenvelopeofthesecoronaviruseshasthreeglycoproteins.ThepeplomersconsistoftheE1membraneglycoproteinwithamolecularmassof24-26kDa,theE2glycoproteinwithamolecular

8、massof100-110kDawhichhascellfusionfunctionsaftercleavagefrom185kDaprecursor[34]andtheE3hemagglutininwithamolecularmassof65kDaoritsdimerof126kDa.Acetylesteraseactivityisassociatedwiththishemagglutinin[3