CpG ODN對新城疫LaSota活疫苗的免疫增強效應.pdf

1、Y774861分類號；密級：p74867忉74880飛媛差蓮夭等。碩士學位論文單位代碼：10019學號：S02316CpGODN對新城疫LaSota活疫苗的免疫增強效應lmmunopotentiationEfficiencyofCpGODNonLaSotaLiveVaccineagainstNewcastleDiseaseinChickens研究指導教生：奎查合作指導教師：塾鑫副熬握申請學位門類級別：壅堂亟稱：亟隨獸匡堂院：動物醫(yī)堂院二

2、oo五年五月名方學業(yè)究在專研所AbstractUnmethylatedCpGdinucleotideswithinspecificflankingbasesarerelativelyabundantinbacterialDNAandareknowntostronglystimulateinnateimmunityinvertebratesTheseimmunostimulatoryactivitiesofbacterialCpGDNAc

3、analsobeachievedwithsyntheticCpGoligodeoxynucleotides(CpGODN)InthisstudytheimmunopoIen“ationefficiencyofsyntheticCpGODNsandrecombinantplasmidscontainingvariouscopiesofCpG2006onLaSotalivevaccineagainstNewcastledisease(ND)

4、wasinvestigatedScreeningCpGODNwithstrongimmunostimulatoryactivityonchickeninvitrowasfirstlycarriedoutusing6sequencesofphosphorothioateCpGODNsdesignedandreferenced，accordingtotheeffectsspecificityandtherelationshipsbetwee

5、nfunctionandstructureofCpGODNTheresultsshowedthatCpG2006，whosesequencewas5’TCGⅡ塹T7FTTGTC_GGTTlWGl要QTT3’，significantlyactivatedBcellstosecreteIgM(p‘O05)，promotedspleniclymphocyteproliferativeactivity(pO01)，increasedmacrop

6、hageNOproduction(p001)，andupregulatedmRNAexpressionofIFNY，IL6andIL1Dinperipheralbloodmononuclearcells(PBMC)(p(005)Moreoverourdataindicatedapoly(G)runatthe3’terminusofCpGODNcouldenhancetheabilityforcellularuptakeandbindin

7、gofCpGODN，andthenupregulateIFN吖levelsecretedbyNKcells，TwosuccessivevaccinationswithLaSotalivevaccinejncombinationoftheseODNswerecarriedoutinchickensinattempttoanalyzetheadjuvanteffectof“CpGODNThreeweekspostboosterimmuniz

8、ation(PBI)，birdsinCpG2006grouppresentedmeanserumtliantibodytiterof821092、lymphoproliferafivestimulationindexof9836，macrophageNOproductionof35833pmol／L，showingasignificantincrementby21092(p‘O05)，44(po01)，276gmol／L(p001)fr

9、omthecontrolgroup，respectivelyFurthermore，ahighrelativeamountofcytokinemRNAexpressionforIFNT、IL一6andIL一1pandaprotectionrateof100％fromNDVchallengewereobservedinCpG2006groupThedataindicatedtherewereremarkabledifferencesbet

10、weenCpG2006groupandthecontrolgroup(p001)TheseresultsdemonstrateCpG2006isabletonotablyenhancespecificantibodyresponsesandstimulateastrongandbalancedThllTh2cellularimmuneresponsetoLaSotalivevaccine，suggestingthatCpG2006mig

11、hthavepotentialasanadjuvantoflivevaccineagainstNDTwostringsofCpGDNAcontaining8andI6copiesofCpG2006monomerwereobtainedrespectivelythroughspliceoverlappingextensionPCR(SOEPCR)andTAcloningThetwotargetCpGDNAfragmentswerethen

12、clonedintopUCl9respectivelytoconstructtwecombinantplasmids，pUCCpG8andpUCCpGl6Toinvestigateadjuvanteffectoftherecombinantplasmids，chickenswereimmunizedrepeatedlywithLaSotalivevaccinecombinedwiththetwoplasmidsThreeweeksPBI