简介：Theimmuneefficiencyofarecombinantadenovirustype5withtype35fibercontainingHIV-1gaggene(rAd5/F35-mod.gag)wasinvestigatedinBALB/cmice,inwhichtherAd5/F35-mod.gagwasfirstlyidentifiedwithPCR,thentransfectedto293cellsandtheinvitroexpressionlevelofGagproteinwasdeterminedbyWesternblottingandindirectimmuno-fluorescentassay.MicewereimmunizedwithintramuscularinjectionsofrAd5/F35-mod.gag,rAd5-mod.gagorDNAandwereboostedafter3weeks.Totesttheeffectofpre-existinganti-viralimmunityonimmunization,micewerealsoinjectedwithAd5-GFPvectorandthenimmunized4and7weekslaterwithAd5/F35-mod.gagvector.TheP24-specificIgGantibodyinseraofimmunizedmicewasdeterminedbyELISAandthespecificcytotoxicTlymphocyte(CTL)responsewasassayedbyintracellularcytokinestaining.ItwasdemonstratedthattherAd5/F35-mod.gagvectorcouldexpressefficientlytheHIVGagproteinin293cellsinvitroandinducestrongHIV-specificimmuneresponsesinvivo.ThestrongestCTLandserumIgGresponseoccurredwhenmicewereimmunizedtwicewithinjectionofrAd5/F35alone,buttheanti-Ad5antibodyafterprimaryinfectionwithadenoviruscouldinhibitthespecificimmuneresponsesinducedbyrAd5/F35vector.ItisconcludedthatsingleimmunizationwithrecombinantadenovirusrAd5/F35-mod.gagcaninducespecificCTLandserumIgGantibodyresponsesinmice,buttheimmunogenicityofrAd5/F35iscomparablyweakerthanthatofrAd5.

简介：ToinvestigatethephenotypicknockoutofHIV-1chemokinecoreceptorCXCR4andCCR5byintrakinesanditsinhibitoryeffectonHIV-1infection.PrimaryhumanPBLsweretransducedwiththerecombinantvectorpLNCX-R-K-S-K(△NGFR),followedbyanti-NGFR/anti-IgG-magneticbeadmethodselectionandFCMdetection.ThetransducedPBLswereinfectedwithDP1HIV-1virusthereafterenvelope-mediatedsyncytiumformationandp24detectionwerecarriedouttostudytheblockageofHIV-1infectionbyco-inactivationofCCR5andCXCR4.pLNCX-R-K-S-K(△NGFR)-transducedPBILswereisolatedwithananti-NGFR/anti-IgG-magneticbeadmethod.Afterisolation,about70%ofthePBLswerepositivefortheNGFRmarker.WhenthetransducedPBLswereinfectedwithDP1HIV-1virus,envelop-mediatedsyncytiumformationwasalmostcompletelyinhibitedbypLNCX-R-K-S-K(△NGFR)transfection.Also,p24antigenwasverylowintheculturesofpLNCX-R-K-S-K(△NGFR)transducedPBLs.pLNCX-R-K-S-K(△NGFR)transductioninhibitedtheproductionofDP1p24antigenby15%,43%and19%ondays4,7and10respectively.ThelymphocyteswiththephenotypicknockoutofCCR5andCXCR4couldprotectprimaryhumanPBLsfromDP1HIV-1virusinfection.