Bacterial Antigen Expression Is an Important Component in Inducing an Immune Response to Orally Administered Salmonella-Delivered DNA Vaccines 英文参考文献.docVIP

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Bacterial Antigen Expression Is an Important Component in Inducing an Immune Response to Orally Administered Salmonella-Delivered DNA Vaccines 英文参考文献.doc

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Bacterial Antigen Expression Is an Important Component in Inducing an Immune Response to Orally Administered Salmonella-Delivered DNA Vaccines 英文参考文献

BacterialAntigenExpressionIsanImportantComponent inInducinganImmuneResponsetoOrallyAdministered Salmonella-DeliveredDNAVaccines MichelleE.Gahan1,2,3¤*,DianeE.Webster2,4,StevenL.Wesselingh2,3,RichardA.Strugnell1,JiYang1 1DepartmentofMicrobiologyandImmunology,UniversityofMelbourne,Parkville,Victoria,Australia,2Children’sVaccinesGroup,MacfarlaneBurnetInstituteforMedical ResearchandPublicHealth,Melbourne,Victoria,Australia,3DepartmentofMedicine,MonashUniversity,AlfredHospital,Prahran,Victoria,Australia,4SchoolofBiological Sciences,MonashUniversity,Victoria,Australia Abstract Background:TheuseofSalmonellatodeliverheterologousantigensfromDNAvaccinesisawell-acceptedextensionofthe successoforalSalmonellavaccinesinanimalmodels.AttenuatedS.typhimuriumandS.typhistrainsaresafeandefficacious, and their use to deliver DNA vaccines combines the advantages of both vaccine approaches, while complementing the limitations of each technology. An important aspect of the basic biology of the Salmonella/DNA vaccine platform is the relative contributions ofprokaryotic andeukaryotic expressionin production ofthevaccine antigen.Gene expressionin DNAvaccinesiscommonlyunderthecontroloftheeukaryoticcytomegalovirus(CMV)promoter.Theaimofthisstudywas to identify and disable putative bacterial promoters within the CMV promoter and evaluate the immunogenicity of the resultingDNAvaccinedeliveredorallybyS.typhimurium. Methodology/PrincipalFindings:Theresultsreportedhereclearlydemonstratethepresenceofbacterialpromoterswithin theCMVpromoter.Thesepromotershavehomologytothebacterialconsensussequenceandfunctionalactivity.Todisable prokaryoticexpressionfromtheCMVpromoteraseriesofgeneticmanipulationswereperformedtoremovethetwomajor bacterialpromotersandaddabacteriatranscriptionterminatordownstreamoftheCMVpromoter.S.typhimuriumwasused toimmuniseBALB/cmiceorallywithaDNAvaccineencodingtheC-fragmentoftetanustoxin(TT)undercontrolofthe originalorthemodifiedCMVpromoter.Althoughbothpromotersfunctionedequallywellin