Establishment of a Reverse Genetics System for Studying Human Bocavirus in Human Airway Epithelia 英文参考文献.docVIP

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Establishment of a Reverse Genetics System for Studying Human Bocavirus in Human Airway Epithelia 英文参考文献.doc

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Establishment of a Reverse Genetics System for Studying Human Bocavirus in Human Airway Epithelia 英文参考文献

EstablishmentofaReverseGeneticsSystemforStudying HumanBocavirusinHumanAirwayEpithelia QinfengHuang1.,XuefengDeng1,2.,ZiyingYan3.,FangCheng1,YongLuo1,WeiranShen1, DianaC.M.Lei-Butters3,AaronYunChen1,YiLi2¤,LiangTang4,MariaSo¨derlund-Venermo5, JohnF.Engelhardt3,JianmingQiu1* 1DepartmentofMicrobiology,MolecularGeneticsandImmunology,UniversityofKansasMedicalCenter,KansasCity,Kansas,UnitedStatesofAmerica,2CollegeofLife Sciences,CentralChinaNormalUniversity,Wuhan,China,3DepartmentofAnatomyandCellBiology,CollegeofMedicine,UniversityofIowa,IowaCity,Iowa,United StatesofAmerica,4DepartmentofMolecularBiosciences,UniversityofKansas,Lawrence,Kansas,UnitedStatesofAmerica,5HaartmanInstitute,DepartmentofVirology, UniversityofHelsinki,Helsinki,Finland Abstract Humanbocavirus1(HBoV1)hasbeenidentifiedasoneoftheetiologicalagentsofwheezinginyoungchildrenwithacute respiratory-tractinfections.Inthisstudy,wehaveobtainedthesequenceofafull-lengthHBoV1genome(includingboth termini) using viral DNA extracted from a nasopharyngeal aspirate of an infected patient, cloned the full-length HBoV1 genome,anddemonstratedDNAreplication,encapsidationofthessDNAgenome,andreleaseoftheHBoV1virionsfrom human embryonic kidney 293 cells. The HBoV1 virions generated from this cell line-based production system exhibits a typicalicosahedralstructureofapproximately26nmindiameter,andiscapableofproductivelyinfectingpolarizedprimary humanairwayepithelia(HAE)fromtheapicalsurface.InfectedHAEshowedhallmarksoflungairway-tractinjury,including disruptionofthetightjunctionbarrier,lossofciliaandepithelialcellhypertrophy.Notably,polarizedHAEculturedfroman immortalizedairwayepithelialcellline,CuFi-8(originallyderivedfromacysticfibrosispatient),alsosupportedproductive infection of HBoV1. Thus, we have established a reverse genetics system and generated the first cell line-based culture systemforthestudyofHBoV1infection,whichwillsignificantlyadvancethestudyofHBoV1replicationandpathogenesis. Citation:HuangQ,DengX,YanZ,ChengF,Luo