表達(dá)豬圓環(huán)病毒2型衣殼蛋白重組質(zhì)粒的構(gòu)建及免疫效應(yīng).pdf

1、Y773593分類號(hào)：密級(jí)：’專蛾差弋夭普博士學(xué)位論文單位代碼：10019學(xué)號(hào)：B0221lII表達(dá)豬圓環(huán)病毒2型衣殼蛋白重組質(zhì)粒r。；≯的構(gòu)建及免疫效應(yīng)ConstructionandImmuneEfficacyofRecombinantPlasmidsExpressingCapsidProteinofPorcineCireovirusType2研究指導(dǎo)教師：疊墊盤熬控合作指導(dǎo)教師：申請(qǐng)學(xué)位門類級(jí)別：壅生譴專業(yè)名稱：亟睦蘭匡莖研究方向：

2、盤筮僉主痘圭芏盞塞盎鱟所在學(xué)院：疊塹匡堂瞳2005年5月AbstractPorcinecircovirustype2(PCV2)istheprimarypathogenofpostweaningmultisystemiewastingsyndrome(PMWS)andotherassociatedconditionsinpigsInthisstudyseveralmammalianrecombinantplasmids，expressi

3、ngporcinegranulocyte—macrophagecolony—stimulatingfactor(GMCSF)，PCV2capsidprotein(cP)，andCOexpressingCPandGMCSFwereconstructedrespectivelyThehumeralendcellularimmuneresponseselicitedbyrecombinantplasmidsexpressingPCV2CPwe

4、reanalyzedinmiceandpigs；moreoverregulatingfunctionsofrecombinantplasmidsexpressingporcineGMCSFandIL2toPCV2DNAvaccine，respectivelywasevaluatedatthesametimeThecDNAofporeineGMCSFgeneWasamplifiedfromthepefipheralbloodlymphoc

5、ytes(PBL)ofswinebyRrrPCRandWasusedtoconstructarecombinanteukaryoticexpressionplasmidpcDNA3，1gmcsferr2geneofPCV2wasobtainedbyPCRandthreerecombinantplasmidsincludingPeDNA3，1off2pcDNA4err2andplRESorf2／gmcsfCO—expressingCP＆G

6、MCSFwereconstructedrespectivelyItwastestifiedthatCPgeneinpcDNA31err2，pcDNA4orf2orplRESorf2／gmcsfGMCSFgeneinpcDNA31gm—csfandplRESorf2lgmesfcouldbeexpressedinCOS7cellsbyindirectimmunofluoresceneassayandcolonystimulationfor

7、mingassayThehumoralendcellularimmuneresponsesinducedbytherecombinantplasmidspcDNA31一orf2andpcDNA4orf2wereanalyzedinmiceTheresultsshowedthatafterthreeintramuscularinjections，specificantibodiestoPCv2werepresentinserumof80％

8、micewithpeDNA31一off2and100％micewithpcDNA4orf2；thesplenocytesproliferationofmiceimmunizedwithrecombinantplasmids，stimulatedbyconcanavalinA(ConA)orrecombinantPCV2cPweredistinctlyincreased，andthetysisactivitiesofCTLandNKcel

9、lsweresignificantlyelevated；peDNA4一ort2aroserelativelylowercellularimmuneresponsesincomparisonwithpeDNA31off2recombinantplasmids；thenumberofCD3CD4CD8andCD3CD4CDSTlymphocytessub—populationsofsplenocytesroseafterthemicewer

10、eimmunizedwithrecombinantplasmids；OntheotherhandtherewerenodifferencesinimmuneresponsesinducedbyintramuscularandintranasaladministrationofpcDNA31一orf27dayoldhealthypigletswererandomlydividedinto5groupswhichWereblankvecto

11、r(groupD，pcDNA31orf2(groupⅡ)，PeDNA31orf2pluspcDNA31gmesf(group11I)plRESorf2／gmcsf(groupIV)andpcDNA31orf2pluspcDNA31IL2(encodingporcineinterlukin2)(groupv)Allpigletswereinjectedintramuscularlythreetimesat2weekintervalsThe

12、resultsshowedthatallgroupsofpigsimmunizedwithrecombinantplasmidscouldproducelowlevelspecificELISAantibodytoPCV2andneutralizingantibodieswithaaveragetitre1：67(groupII)，1：293(group111)，1：267(groupIV)and1：202(groupV)respect

13、ively；theproliferationactivitiesofPBLfrompigsinGroupIVandVwerehiigherthanthoseofpigsinGroup11and1II；meanwhile，lysisactivitiesofNKcellsofpigsinGroup111，IVandVclearlyexceededGroup11；thenumberofThTe，andm／aThofpigsineachgrou