Chronic Wasting Disease in Bank Voles Characterisation of the Shortest Incubation Time Model for Prion Diseases.docVIP

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Chronic Wasting Disease in Bank Voles Characterisation of the Shortest Incubation Time Model for Prion Diseases.doc

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Chronic Wasting Disease in Bank Voles Characterisation of the Shortest Incubation Time Model for Prion Diseases

ChronicWastingDiseaseinBankVoles:Characterisation oftheShortestIncubationTimeModelforPrionDiseases MicheleAngeloDiBari1*,RomoloNonno1,Joaqu?′nCastilla2,ClaudiaD’Agostino1,LauraPirisinu1, GeraldinaRiccardi1,MichelaConte1,JuergenRicht3¤,RobertKunkle3,JanLangeveld4,GabrieleVaccari1, UmbertoAgrimi1 1DepartmentofVeterinaryPublicHealthandFoodSafety,IstitutoSuperiorediSanita`,Rome,Italy,2CICbioGUNEandIKERBASQUE,BasqueFoundationforScience,Derio andBilbao,Bizkaia,Spain,3NationalAnimalDiseaseCenter,AgriculturalResearchService,UnitedStatesDepartmentofAgriculture,Ames,Iowa,UnitedStatesofAmerica, 4DepartmentofInfectionBiology,CentralVeterinaryInstituteofWageningenUR,Lelystad,Netherlands Abstract Inordertoassessthesusceptibilityofbankvolestochronicwastingdisease(CWD),weinoculatedvolescarryingisoleucine ormethionineatcodon109(Bv109IandBv109M,respectively)withCWDisolatesfromelk,muledeerandwhite-taileddeer. Efficienttransmissionrate(100%)wasobservedwithmeansurvivaltimesrangingfrom156to281dayspostinoculation. SubsequentpassagesinBv109IallowedustoisolatefromallCWDsourcesthesamevole-adaptedCWDstrain(Bv109I CWD), typifiedbyunprecedentedshortincubationtimesof25–28daysandsurvivaltimesof,35days.Neuropathologicaland 109I molecular characterisation of Bv CWD showed that the classical features of mammalian prion diseases were all recapitulatedinlessthanonemonthafterintracerebralinoculation.Bv109ICWDwascharacterisedbyamildanddiscrete distributionofspongiosisandrelativelylowlevelsofprotease-resistantPrPSc(PrPres)inthesamebrainregions.Despitethe low PrPres levels and the short time lapse available for its accumulation, end-point titration revealed that brains from terminally-illvolescontainedupto108,4i.c.ID50infectiousunitspergram.Bv109ICWDwasefficientlyreplicatedbyprotein misfoldingcyclicamplification(PMCA)andtheinfectivityfaithfullygeneratedinvitro,asdemonstratedbythepreservation ofthepeculiarBv109ICWDstrainfeaturesonre-isolationinBv109I.Overall,weprovideevidencethatthesameCWDstrain was iso