Replication of Epstein-Barr Virus Primary Infection in Human Tonsil Tissue Explants 英文参考文献.docVIP

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Replication of Epstein-Barr Virus Primary Infection in Human Tonsil Tissue Explants 英文参考文献.doc

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Replication of Epstein-Barr Virus Primary Infection in Human Tonsil Tissue Explants 英文参考文献

ReplicationofEpstein-BarrVirusPrimaryInfectionin HumanTonsilTissueExplants KenseiGotoh1,YoshinoriIto1*,SeijiMaruo2,KenzoTakada2,TerukazuMizuno3,MasaakiTeranishi3, SeiichiNakata3,TsutomuNakashima3,SeikoIwata4,FumiGoshima4,ShigeoNakamura5 ,Hiroshi Kimura4 1DepartmentofPediatrics,NagoyaUniversityGraduateSchoolofMedicine,Nagoya,Japan,2DepartmentofTumorVirology,InstituteforGeneticMedicine,Hokkaido University,Sapporo,Japan,3Department ofOtorhinolaryngology,NagoyaUniversityGraduate SchoolofMedicine,Nagoya,Japan, 4DepartmentofVirology, Nagoya UniversityGraduateSchoolofMedicine,Nagoya,Japan,5DepartmentofPathologyandLaboratoryMedicine,NagoyaUniversityHospital,Nagoya,Japan Abstract Epstein-Barrvirus(EBV)maycauseavarietyofvirus-associateddiseases,butnoantiviralagentshaveyetbeendeveloped against this virus. Animal models are thus indispensable for the pathological analysis of EBV-related infections and the elucidationoftherapeuticmethods.ToestablishamodelsystemforthestudyofEBVinfection,wetestedtheabilityofB95– 8virusandrecombinantEBVexpressingenhancedgreenfluorescentprotein(EGFP)toreplicateinhumanlymphoidtissue. Humantonsiltissuesthathadbeensurgicallyremovedduringroutinetonsillectomyweresectionedintosmallblocksand placedontopofcollagenspongegelsinculturemediumattheair-interface,thenacell-freeviralsuspensionwasdirectly appliedtothetopofeachtissueblock.IncreasinglevelsofEBVDNAinculturemediumwereobservedafter12–15days through 24 days post-infection in tissue models infected with B95–8 and EGFP-EBV. Expression levels of eight EBV- associatedgenesincells collectedfromculturemediumwereincreasedduringculture.EBV-encodedsmallRNA-positive cellsweredetectedintheinterfollicularareasinparaffin-embeddedsections.Flowcytometricanalysesrevealedthatmost EGFP cellswereCD32CD562CD19 HLA-DR+,andrepresentedbothna?¨ve(immunoglobulinD )andmemory(CD27+ )B cells.Moreover,EBVreplicationinthismodelwassuppressedbyacyclovirtreatmentinadose-dependentmanner.These data suggest that this model ha