Anoveltuberculosis(TB)genevaccinecontainingmousegranulocytemacrophage-colonystimulatingfactor(mGM-CSF)andaTBantigen(Ag85A)wasdevelopedinthisstudy.ThegenesencodingAg85AandmGM-CSFwereamplifiedbyPCRrespectivelyfromtheAg85A-containingpBSby5andpC-mGM-CSF.ThegeneswerethenclonedintotwodifferentpolylinkersitesofplasmidpIRES,forminganovelTBgenevaccineconstructpI85AGM.FollowingtransfectionofpI85AGMplasmidinto7721celllinebyLipofectamineTM,theexpressionofAg85AandGM-CSFproteinswasidentifiedbyWesternblottingorRT-PCR.ThenBalb/cmicewereinoculatedwiththerecombinantpI85AGM,pI85A,pIGMorplasmidalone,respectively.TheactivitiesofCTL,NKcellsandtheAg85A-stimulatedproliferationofspleencellsweremeasuredbyMTTmethod.TheserumantibodyagainstAg85AwasdetectedbyELISA.TheresultsshowedthattheAg85AandGM-CSFproteinscouldbeexpressedin7721celllineandtheactivityofCTLsandtheproliferationofspleencellsweresignificantlyincreasedinthepI85AGM-immunizedmice,indicatingthatthepI85AGM-immunizedmicecouldgeneratespecificimmuneresponsestoAg85A.Thisstudymightprovidepossibilityfordevelopingnovelanti-TBgenevaccine.
