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1、JournalofClinicalVirology12(1999)43±51Adaptationofhumanentericcoronavirustogrowthincelllinesa,bcJ.amesP.Luby*,R.ebeccaClinton,S.tanleyKurtzaUTSouthwesternMedicalCenter,Di6isionofInfectiousDiseases,5323HarryHinesBl6d,Dallas,TX75235-9113,USAb6102Parliament,Austin,TX78724,USAc4129RowanDri6e,FortWor
2、th,TX76116,USAReceived14July1998;receivedinrevisedform14October1998;accepted21October1998AbstractBackground:Theexistenceofhumanentericcoronavirus(HEC)hasbeendebatedsinceits?rstdescriptioninstoolbyelectronmicroscopy(EM)in1975.Neededtoresolvetheissueisitscultivationinreadilyavailablecelllines.Obje
3、ctives:TogrowHECincelllines.Todescribeitscharacteristicsandtodifferentiateitfromotherhumanandanimalcoronaviruses.Studydesign:Originallygrowninhumanfetalintestinalorganculture,HECwaspassedinJ774cells(amousemacrophagecellline)andC6:36cells(amosquitocellline).Itscytopathiceffect(CPE)andpatternofimm
4、unoˉuores-cenceweredescribed.ItsappearancewasascertainedbynegativestainingandtransmissionEM.Itsstructuralproteinsweredelineatedbypolyacrylamidegelelectrophoresis(PAGE)andWesternblotting(WB).TheantigeniccharacteroftheviruswasdeterminedbyimmunoˉuorescenceandWB.Agglutinationwithmouseerythrocyteswas
5、performed.Results:InJ774cells,HECinducedtheformationofgiantcellsandsmallsyncytia.ImmunoˉuorescenceinbothJ774andC6:36cellswaslimitedtothecytoplasm.StudieswithtransmissionEMrevealedthevirustohavethetypicalappearanceofothercoronaviruses,tobe80±120nmindiameter,andtobudintocysternaeoftheendoplasmicre
6、ticulum.ByPAGEandWB,itsmajorproteinhasanaveragemolecularweight(MW)of41kilodaltons(kDa).TwootherproteinshadMWsof190and24kDa.ByimmunoˉuorescenceandWB,HECisantigenicallydistinctfromhumancoronaviruses0C43and229Eandmousehepatitisvirus(A59strain).PreparationsofHECdidnotagglutinatemouseerythrocytes.Con
7、clusion:WeconcludethatHECisahumancoronavirusthatisantigenicallyunrelatedto0C43and229Eviruses.GrowthofHECinreadilyavailablecelllinesshouldaidinelucidatingitsroleasapathogeninhumandiarrhealillnesses.?1999ElsevierScienceB.V.All
