Skin Electroporation Effects on Transgene Expression, DNA Persistence and Local Tissue Environment 英文参考文献.docVIP

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Skin Electroporation Effects on Transgene Expression, DNA Persistence and Local Tissue Environment 英文参考文献.doc

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Skin Electroporation Effects on Transgene Expression, DNA Persistence and Local Tissue Environment 英文参考文献

SkinElectroporation:EffectsonTransgeneExpression, DNAPersistenceandLocalTissueEnvironment Anna-KarinRoos1¤*,FredrikEriksson1,JamesA.Timmons2,JosefineGerhardt1,UlrikaNyman3 ,Lindvi Gudmundsdotter4,AndreasBra?ve4,BrittaWahren4,PavelPisa1 1Department of Oncology and Pathology, Cancer Center Karolinska R8:01, Immune and Gene Therapy Laboratory, Karolinska Institute, Stockholm, Sweden, 2Royal VeterinaryCollege,UniversityofLondon,Camden,London,UnitedKingdom,3InstituteofEnvironmentalMedicine,ToxicologyandNeurotoxicology,KarolinskaInstitute, Stockholm,Sweden,4DepartmentofMicrobiology,TumorandCellBiology,KarolinskaInstitutetSwedishInstituteforInfectiousDiseaseControl,Stockholm,Sweden Abstract Background:Electricalpulseshavebeenusedtoenhanceuptakeofmoleculesintolivingcellsfordecades.Thistechnique, oftenreferredtoaselectroporation,hasbecomeanincreasinglypopularmethodtoenhanceinvivoDNAdeliveryforboth gene therapy applications as well as for delivery of vaccines against both infectious diseases and cancer. In vivo electrovaccination (gene delivery followed by electroporation) is currently being investigated in several clinical trials, includingDNAdeliverytohealthyvolunteers.However,themodeofactionatmolecularlevelisnotyetfullyunderstood. Methodology/Principal Findings: Thisstudy investigates intradermal DNA electrovaccination in detail anddescribes the effectsonexpressionofthevaccineantigen,plasmidpersistenceandthelocaltissueenvironment.Geneprofilingofthe vaccination site showed that the combination of DNA and electroporation induced a significant up-regulation of pro- inflammatorygenes.Invivoimagingofluciferaseactivityafterelectrovaccinationdemonstratedarapidonset(minutes)and alongduration(months)oftransgeneexpression.However,whenthemoreimmunogenicprostatespecificantigen(PSA) wasco-administered,PSA-specificTcellswereinducedandconcurrentlytheluciferaseexpressionbecameundetectable. Electroporationdidnotaffectthelong-termpersistenceofthePSA-expressingplasmid. Conclusions/Signific