简介：ToclarifytheroleofAPOBEC3G(A3G)incellulardefenseagainsthepatitisBvirus(HBV),theexpressionofA3GinnormalhumanliverandtheregulationoftheA3Gexpressioninhep-atomacellline(HuH-7)wereinvestigated.ExpressionlevelofAPOBEC3smRNAinhumanliverwasdeterminedbyRT-PCR.HuH-7andHepG2cellsweretreatedwithvariousconcentrationsofIFN-α(0U/ml,100U/ml,500U/ml,1000U/ml)for12h.ThemRNAlevelsweremeasuredbyaquantitativeRT-PCR,theresultswerenormalizedrelativetothespecimenswithoutIFN-αstimulation.TotalproteinofHuH-7cellstreatedwithvariousconcentrationsofIFN-αfor48hwassubjectedtoWesternblotanalysis.Forreportergeneassay,HuH-7cellsweretransfectedwiththereporterplasmidscontainingIRF-Esitesanditsmutantswithdifferentlengths.Thenthecellsweretreatedwithorwithout1200U/mlIFN-aforadditional12h(1000U/ml)after24hoftransfection,andthecelllysatewaspreparedandassayedforluciferaseactivity.ItwasfoundthatnormalhumanliverexpressedthemRNAofA3G.A3GmRNAexpressioninHuH-7andHepG2cellswereup-regulatedbyIFN-αstimulationinadose-depen-dentmanner.WesternblotanalysisindicatedthatA3GproteinexpressionwasalsoenhancedbyIFN-αstimulation.SequenceanalysisshowedtheexistenceofputativesitesofIFNregulatoryfactorelement(IRF-E)in5'regionofA3Ggeneupstreamtheinitiationcodon.IFN-αstimulationresultsin6-to8-foldincreaseinluciferaseactivityincellstransfectedwiththeplasmidcontainingIRF-Esitesofthe5'upstreamsequences,whereasluciferaseactivitydidnotchangeincellstransfectedwiththeplasmidcontainingmutantIRF-EsitesorwithoutIRF-Esites.Asaconclusion,A3Gareexpressedinnormalhumanliver.A3Gexpressionwasup-regulatedbyIFN-αstimulationinhepatomacellsandcouldbeinvolvedinhostdefensemechanismsagainstHBV.ERF-Esitein5'regionofAP0BEC3Ggeneupstreamtheinitiationcodonplaysanimportantroleinthisp

简介：TheaimofthepresentstudywastodeterminetheefficacyofimmunotherapywithdendriticcellstoelicitEBV-specificCTL-immunityinadvancedcasesofEBV-positivepatientswithnasopharyngealcarcinoma(NPC)andtodeterminethesafetyandtoxicityofthispreparation.NinecasesofhistologicallyconfirmedpatientswithNPCundergoingtreatmentwithradiologicaltherapywereenrolledinthisstudy.Dendriticcells,generatedinvitrofrombloodmonocytesofpatientswereculturedandmaturedwithcytokinesandtheninfectedwithrecombinantadenovirusvaccinecontainingEBV-latentmembraneprotein-2(Ad-LMP2).On9days'cultivationofcells,thematuredDCswereharvested,irradiatedwithCoandtheninjectedintradermallytopatientswithNPC.Theinjectionswereperformed3timestotally.Afterimmunization,theCTLresponseswereassayedbymeansofcytotoxicityandepitope-specificIFN-γproduction.Theresultsofthistrialshowedthatallpatientscouldtoleratethiskindoftreatmentwithoutanysideeffect,duringwhichmarkedincreaseofLMP2-specificCTL-responsescouldbedemonstratedin5patientsofthisgroup.AndthelevelofIgA/VCAantibodydecreasedin8of9patients,thusaccountingforabetterprognosisforthesepatients.Allpatientswillbefollowedupforanotheroneyear.Atleast,thepresentworkshowsthatintradermalvaccinationwithautologousDCsinfectedwithrecombinantAd-LMP2adenovirusisasafeprocedureinNPCpatients,inwhichthisprocedurecanenhancetheLMP2-specificCTLresponsesinpatients.Thesedataareencouragingtodevelopmoreeffectivevaccinestrategiesforthetreatmentofnasopharyngealcarcinoma.

简介：Toinvestigatetheinfluenceofmycophenolatemofetil(MMF)uponthematurationandtheallo-stimulatoryactivityofculturedprogenitorsofdendriticcells(DCp),andtoevaluatetheeffectsofthepre-treateddentriticcellsofrecipientswithMMFonthetoleranceinductionaswellasitspossiblemechanism,GM-CSFandMMFwereaddedtotheinvitroculturedprogenitorcells,andtheimmuno-phenotypicalanalysiswasperformedbymeansofflowcytometry.ThesecretionofIL-12wasdetectedbyELISAandthestimulatoryactivitiesofDCponallogeneicTcellswereobservedbymixedlymphocytereaction.Twenty-fourC57BL/6miceweredividedinto3groups(eachwith8mice),inwhichgroupAofmiceacceptedallograftsofheartfromBALB/cmice,groupBofmicehadreceiveduntreatedDCpfromdonorsofBALB/cmice7daysbeforetransplantation,andC57BL/6miceingroupCweretreatedbyinjectionwithMMF-treatedallograftsofheartfromBALB/cmice7daysbeforetransplantation.Thesurvivaltimesofallograftsandthechangesofthecytokinelevelsinsereoftherecipientmicewereobservedaftertransplantation.TheexperimentalresultsshowedthatMMFcouldsignificantlyinhibittheexpressionsofthecostimulatorymoleculesCD80andCD86onDCsandthesecretionofIL-12andtheallo-stimulatoryactivitiesofDCswerealsomarkedlyinhibited.ThesurvivaltimesofallograftsingroupBofmicewerelongerthanthoseingroupA,whilethegroupCshowedthelongestsurvivaltimesofallografts,withamarkedreductionintheproductionoftheThltypecytokines.ItisevidentthatMMFhasasuppressiveeffectonthematurationandallo-stimulatoryactivitiesofthecultureddendriticcellprogenitors,thusleadingtoadonorspecifictoleranceinheart-transplantedrecipients.