表达融合蛋白cfp21-mpt64和esat-6-cfp10dna疫苗增强bcg初免小鼠抗结核病保护性-dna vaccine expressing fusion protein cfp21 - mpt64 and esat - 6 - cfp10 enhance the anti-tuberculosis protection of bcg primary immunized mice..docxVIP

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2018-05-28 发布于上海
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表达融合蛋白cfp21-mpt64和esat-6-cfp10dna疫苗增强bcg初免小鼠抗结核病保护性-dna vaccine expressing fusion protein cfp21 - mpt64 and esat - 6 - cfp10 enhance the anti-tuberculosis protection of bcg primary immunized mice..docx

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表达融合蛋白cfp21-mpt64和esat-6-cfp10dna疫苗增强bcg初免小鼠抗结核病保护性-dna vaccine expressing fusion protein cfp21 - mpt64 and esat - 6 - cfp10 enhance the anti-tuberculosis protection of bcg primary immunized mice.

DNAvaccinesexpressingCFP21-MPT64andESAT-6-CFP10fusionproteinsenhanceBCG-inducedprotectiveimmunityagainstmousetuberculosisBackgroundandaimsAbstractBCGistheonlyavailablevaccineforthepreventionofTBandthecoveragerateisabout90%inthemostdevelopingcountriesworldwild.However,theefficacyofBCGvaccineinpreventingadultTBishighlyvariableandandmechanismsforthishavenotbeenconfirmedyet.Thereare16RDsidentifiedinthegenomeofM.tbH37Rvsofar.TheprimarydeletionofRD1between1908and1921,whichisabsentfromallBCGsubstrainsbutpresentinvirulentM.tuberculosisandM.bovis,isthoughttobethemostobviousreasonfortheattenuationofBCG.Moreover,bythedeletionofRD2regionduring1927and1931,BCGsubstrainsaredividedintoearlysubstrains(BCGJapan,Russia,Moreau,Sweden,andBirkhaug)andlatesubstrains(BCGTice,Frappier,Phipps,Prague,Danish,Pasteur,Glaxo).RecentstudiessuggestthatRD1couldenhancetheprotectiveefficacyofBCGintheformsofrecombinantBCGorsubunitvaccine.TheeffectoflackofRD2ontheprotectiveefficacyofBCGsubstrainsagainstTBremainsunknown.CFP10(Rv3874)andESAT-6(Rv3875),CFP21(Rv1984c)andMPT64(Rv1980c),havebeenconsideredimportantimmunodominantantigensencodedbyRD1andRD2ofM.tuberculosis,respectively.InordertoelucidatetheeffectoflossofRD2ontheefficacyofBCG,theimmuneresponsesgeneratedagainstDNAvaccinesexpressingCFP21-MPT64andESAT-6-CFP10fusionproteinsandtheimmunogenicityofbothfusionproteinsintuberculinskintest(TST)positivehealthypopulationwerecomparedinthepresentstudy.Moreimportantly,wecomparedtheuseofaBCGprime-DNAvaccinesbooststrategytoinduceprotectionagainstvirulentM.tbchallengeinC57BL/6mice.MethodsThecodingsequencesofbothEAST-6andCFP10proteinswereamplifiedbyPCR.Theesat-6-cfp10wasgeneratedbyasecondPCRaccordingtothemethodofgenesplicingwithoverlapextension.ThefusiongenewasinsertedintoprocaryoticexpressionvectorpProEXHTbandeucaryoticexpressionvectorpcDNA3.1(-),resultingintherecombinantplasmidspPro610andpcD610,respectively.Meanwhile,pPro2164wasdigestedwithBamHIandHindIIIandtheninsertedintopcDNA3.1(-),resultingintherecombinantplasm