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《弓形虫棒状体蛋白38的序列变异与免疫保护作用分析》由会员上传分享，免费在线阅读，更多相关内容在行业资料-天天文库。
1、DNAvaccinepVAX-ROP38.AfterpVAX-ROP38wastransfectedintoHEK293cells,indirectimmunofluorescenceassay(IFA)wasusedtodetecttheexpressionofpVAX-ROP38invitro.MiceintheexperimentalgroupwereintramuscularlyinjectedbypVAX-ROP38andthosereceivedpVAXI,PBSandnothingascontrols
2、.Afterthreetimesinjectionattwoweekintervals,allgroupswerechallengedwith1000tachyzoitesofthevirulentT.gondiiRHstrainand10cystsofthePRUstrain,respectively.TheresultsshowedmiceinoculatedwithpVAX-ROP38vaccinehadahigherlevelofIgGantibodiesandIFN-γandIL-2cytokines,a
3、ctivatedTh1-typeimmuneresponses,producedthehigh+-+++-levelofCD3CD4CD8andCD3CD4CD8Tlymphoproliferativeresponse.Inthemodelofacuteinfection,theaveragesurvivaltimeofmiceinpVAX-ROP38group(8.1d±0.75)wasnostatisticallydifferentcomparedtothatinPBS,pVAXIandblankcontrol
4、groupswhichdiedwithin7d.However,inthemodelofchronicinfection,thebraincystreductioninthepVAX-ROP38groupreached76.6%,whichwassignificantlydecreasedincontrasttocontrols.Finally,weevaluatedtheimmuneprotectionofmixedrROP38andrROP18proteinswhichwereencapsulatedin50:
5、50poly(lactide-co-glycolide)(PLG)slow-releasedmicroparticles.WeconstructedPLG-rROP38andPLG-rROP18microparticles.MiceintheexperimentalgroupswereinjectedwithrROP38,PLG-rROP38,rROP18,PLG-rROP18,rROP38-rROP18andPLG-rROP38-rROP18,respectivellyandthosereceivedPBS,PL
6、G-PBSandblankgroupweretakenascontrols.Afterthelastimmunization,humoralandcellularimmunityrelatedindexeswereexaminedasthatforpVAX-ROP38vaccine.Sixweeksafterthelastimmunization,allgroupswerechallengedwith10cystsofthePRUstrain.Theresultsshowedthatimmunizationofmi
7、cewithproteinvaccinescouldelicitahigherlevelofIgGantibodiesandIFN-γandIL-2cytokines,activatedamixedTh1-Th2+-+++-typeimmuneresponses,producedthehighlevelofCD3CD4CD8andCD3CD4CD8Tlymphoproliferativeresponse.Inthemodelofchronicinfection,thenumberofbraincystsdecrea
8、sed81.26%inthePLG-rROP38-rROP18group,comparedwiththatofthecontrols.Inconclusion,thisstudyamplifiedTgROP38genesuccessfully,andrevealedalowlevel(0.2-1.1%)ofsequencevariationamongdiff
