Immune responses induced by a BacMam virus expressing the E2(6)

150M.Lietal./ImmunologyLetters125(2009)145–150

producedsubunitvaccine.Ourstudyisthe rsttoevaluatetheCSFVE2immunogenexpressedfromtheBacMamvirusundertheie1promoter(ashuttlepromoterbetweenmammalianandinsectcells[12]).Incontrast,XuandLiuutilizedaconventionalrecombi-nantbaculoviruswiththetypicalinsectp10promotertoproducetheE2protein.(2)Weassessedcell-mediatedimmunityinducedbytheBacMamvirususingCFSEandWST-8assays,inadditiontoevaluatingtheantibodyresponses,whiletheotherreportonlyevaluatedtheneutralizingantibodiesinducedbytherecombinantbaculovirus.

Insummary,thepresentstudydemonstratedthattherecombi-nantpseudotypedbaculovirusmediatedef cientgenedeliveryandexpressionofCSFVE2proteininbothinsectcellsandmammaliancells.Importantly,weshowedthatdirectvaccinationwiththeBacMamvirusinducedCSFV-speci chumoralandcell-mediatedimmuneresponses.OurstudyprovidessupportingevidencefortheuseoftheBacMamvirusasavectorforgeneticimmunization.Consideringthesafetyandcost-effectivenessofproducinghigh-titerrecombinantbaculovirus,ourvaccinehasahighpotentialfordevelopmentasapracticalandeffectivevaccineagainstCSF.Tofur-thervalidateour ndings,wewillperformvaccinationstudiesandevaluateprotectionfromchallengeinpigsinthefuture.Acknowledgement

ThisworkwassupportedbyagrantfromTheNational863Pro-gramofChina(2006AA10A204).References

[1]Delaney4thWE,IsomHC.HepatitisBvirusreplicationinhumanHepG2

cellsmediatedbyhepatitisBvirusrecombinantbaculovirus.Hepatology1998;28:1134–46.

[2]BoyceFM,BucherNL.Baculovirusmediatedgenetransferintomammaliancells.

ProcNatlAcadSciUSA1996;93:2348–52.

[3]CondreayJP,WitherspoonSM,ClayWC,KostTA.Transientandstablegene

expressioninmammaliancellstransducedwitharecombinantbaculovirusvector.ProcNatlAcadSciUSA1999;96:127–32.

[4]AirenneKJ,HiltunenMO,TurunenMP,TurunenAM,LaitinenOH,Kulomaa

MS,etal.Baculovirus-mediatedperiadventitialgenetransfertorabbitcarotidartery.GeneTher2000;7:1499–504.

[5]LiuX,LiK,SongJ,LiangC,WangX,ChenX.Ef cientandstablegeneexpression

inrabbitintervertebraldisccellstransducedwitharecombinantbaculovirusvector.Spine2006;31:732–5.

[6]AokiH,SakodaY,JukurokiK,TakadaA,KidaH,FukushoA.Induction

ofantibodiesinmicebyarecombinantbaculovirusexpressingpseudora-biesvirusglycoproteinBinmammaliancells.VetMicrobiol1999;68:197–207.

[7]FacciabeneA,AurisicchioL,LaMonicaN.Baculovirusvectorselicitantigen-speci cimmuneresponsesinmice.JVirol2004;78:8663–72.

[8]AbeT,TakahashiH,HamazakiH,Miyano-KurosakiN,MatsuuraY,TakakuH.

Baculovirusinducesaninnateimmuneresponseandconfersprotectionfromlethalin uenzavirusinfectioninmice.JImmunol2003;171:1133–9.

[9]GronowskiAM,HilbertDM,SheehanKC,GarottaG,SchreiberRD.Bac-ulovirusstimulatesantiviraleffectsinmammaliancells.JVirol2000;73:9944–51.

[10]WangS,FangL,FanH,JiangY,PanY,LuoR,etal.Constructionandimmuno-genicityofpseudotypebaculovirusexpressingGP5andMproteinofporcinereproductiveandrespiratorysyndromevirus.Vaccine2007;25:8220–7.

[11]LinM,LinF,MalloryM,ClavijoA.DeletionsofstructuralglycoproteinE2of

classicalswinefevervirusstrainAlfort/187resolvealinearepitopeofmon-oclonalantibodyWH303andtheminimalN-terminaldomainessentialforbindingimmunoglobulinGantibodiesofapighyperimmuneserum.JVirol2000;74:11619–25.

[12]GaoH,WangY,LiN,PengWP,SunY,TongGZ,etal.Ef cientgenedelivery

intomammaliancellsmediatedbyarecombinantbaculoviruscontainingawhispovirusie1promoter,anovelshuttlepromoterbetweeninsectcellsandmammaliancells.JBiotechnol2007;131:138–43.

[13]SunY,LiuDF,WangYF,LiangBB,ChengD,LiN,etal.Generationandef cacy

evaluationofarecombinantadenovirusexpressingtheE2proteinofclassicalswinefevervirus.ResVetSci2009(inpress).

[14]XuCY,ChengT,LuWX,ChenM,WuT,WangY.Mammaliangene-transfer

andexpressionef cienciesofbaculovirusBacV-CMV-EGFPA.ShengWuGongChengXueBao2004;20:73–7.

[15]PengWP.Epitopeidenti cationofclassicalswinefevervirusE2proteinby

phagedisplay.MasterDegreeDissertationofChineseAcademyofAgriculturalSciences;2007.

[16]PengWP,HouQ,XiaZH,ChenD,LiN,SunY,etal.Identi cationofacon-servedlinearB-cellepitopeattheN-terminusoftheE2glycoproteinofclassicalswinefevervirusbyphage-displayedrandompeptidelibrary.VirusRes2008;135:267–72.

[17]PengWP,XiaZH,HouQ,LiN,SunY,TongGZ,etal.Differencesinglycosylation

oftheE2proteinbetweenvirulentShimenstrainandavirulentC-strainofclassicalswinefevervirus.BingDuXueBao2007;23:389–93.

[18]HsuCS,HoYC,WangKC,HuYC.Investigationofoptimaltransductioncondi-tionsforbaculovirus-mediatedgenedeliveryintomammaliancells.BiotechnolBioeng2004;88:42–51.

[19]SunY,XiaZH,LiangBB,PengWP,QiuHJ.DevelopmentofanindirectELISAfor

detectingantibodiesagainstclassicalswinefevervirusbasedonthebaculovirusexpressedE2protein.VetSciChina2008;38:315–9.

[20]EUDiagnosticmanualforclassicalswinefever(CSF)diagnosis:tech-nicalpart(seconddraftmarch2002).http://viro08.tiho-hannover.de/eg/TechnicalPart4.pdf.p.28–35.

[21]ZhaoHP,SunJF,LiN,SunY,XiaZH,WangY,etal.Assessmentofthecell-mediatedimmunityinducedbyalphavirusreplicon-vectoredDNAvaccinesagainstclassicalswinefeverinamousemodel.VetImmunolImmunopathol2009;129:57–65.

[22]MiyamotoT,MinW,LillehojHS.Lymphocyteproliferationresponseduring

Eimeriatenellainfectionassessedbynew,reliablenonradioactivecolorimetricassay.AvianDis2002;46:10–6.

[23]KimSJ,SungHW,HanJH,JackwoodD,KwonHM.Protectionagainstveryviru-lentinfectiousbursaldiseasevirusinchickensimmunizedwithDNAvaccines.VetMicrobiol2004;101:39–51.

[24]XuCY,ChengT,LuWX,ChenM,WuT,WangYB,etal.Mammaliangene-transfer

andexpressionef cienciesofbaculovirusBacV-CMV-EGFPA.ShengWuGongChengXueBao2004;20:73–7.

[25]IkonomouL,SchneiderYJ,AgathosSN.Insectcellcultureforindustrialproduc-tionofrecombinantproteins.ApplMicrobiolBiotechnol2003;62:1–20.

[26]HoY,LinPH,LiuCY,LeeSP,ChaoYC.Assemblyofhumansevereacuteres-piratorysyndromecoronavirus-likepart …… 此处隐藏：5717字，全部文档内容请下载后查看。喜欢就下载吧 ……