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1、JournalofVirologicalMethodsELSEVIERJournalofVirologicalMethods52(1995)301-307OptimizationofinvitrogrowthconditionsforenterotropicmurinecoronavirusstrainsS.R.Compton*,D.F.Winograd,D.J.GaertnerSectionofComparativeMedicine,YaleUniversitySchoolofMedicine,NewHaven,CT06520-8016,U
2、SAAccepted21October1994AbstractEnterotropicmousehepatitisvirus(MHV)strainshavebeendifficulttogrowincellculture.InanattempttodevelopanefficientinvitrocultivationsystemforenterotropicMHVstrains(MHV-RIandMHV-Y),8murinecelllineswereinoculatedwithMHV-RI-orMI-IV-Y-infectedinfantm
3、ouseintestinalhomogenatesandscreenedfortheproductionofcytopathiceffects.MHV-RIandMHV-YconsistentlyproducedcytopathiceffectsonlyinJ774A.lcells.Bothstrainsproducedtitersof>lo6TCID,,/mlinsubsequentpassagesin5774.1cells.MHVstrains-1,-3,-A59,-JHM,-Sand-DVIMalsoproducedhigh-titer
4、viralstocksinJ774A.lcells.ThereforeJ774A.lcellsarethefirstcellsfoundthatsupportthereplicationofthese8enterotropicandrespiratoryMHVstrains.AfterpassageinJ774A.lcells,MI-IV-RIandMHV-Ycouldinfectpreviouslynon-susceptiblecelllines(17Cl-1,CMT-93,N18andNCTC14691,thoughcytopathice
5、ffectswereoftennegligiblewithMHV-RI.MHV-Y,butnotMHV-RI,grewinL2(Percy)cells.UsingL2(Percy)cells,anagaroseoverlayandGiemsastaining,MHV-Ycouldbequantifiedbyplaqueassay.Infantmousebioassays,plaqueassaysandcellcultureinfectionswerecomparedfortheirsensitivityindetectingMI-IV-Yin
6、infectedintestinalhomogenatesandcellsupematants.Keywords:Murinecoronavirus;MousehepatitisvirusstrainRIandY;Enterotropism;J774A.lcell;PlaqueassayMousehepatitisvirus(MHV),asingularnameforseveralmurinecoronaviruses,causesawidespectrumofclinicalsyndromesrangingfromsubclinicalin
7、fectiontoenteritis,hepatitis,encephalitisanddeath.MHVstrainscanbesegregatedintotwobiotypes,basedontheirinitialsiteofreplication:respiratory(polytropic)anden-*Correspondingauthor.Fax:+1(203)7857499.0166-0934/95/$09.5001995ElsevierScienceB.V.AllrightsreservedSSDI0166-0934(94)
8、00161-8302S.R.Comptonetal./JournalofVirologicalMethods52(I995)301-307terotropic(Ba
