简介：ToobtaintherecombinantsolubleproteinoftheextracellularfragmentofhumanTRAILgeneandtoidentifyitsfunctionpreliminarily,thisgenefragmentwasamplifiedfromperipheralbloodmononu-clearcells(PBMC)byRT-PCRandclonedintovectorpGEM-T-Easyforsequenceanalysis.Theexpres-sionvectorpET-30a/TRAILwasthenconstructedbyDNArecombinationmethodwithaHis-taggeneatthefrontoftheTRAILfragment,andtherecombinantproteinwasexpressedinE.coliBL21(DE3).Meanwhile,theexpressedtargetproteinwaspurifiedwithNi-NTAchromatographycolumnandidentifiedbySDS-PAGEandWesternblotting.TheproliferationinhibitionactivityofTRAIL-HiswasdetectedbyMTTassay.PIstainingandWright-GiemsastainingwereusedtodetectthepresenceoftheTRAIL-in-ducedcellapoptosis.ItwasdemonstratedthatthetargetproteinexpressedinE.coliBL21showedthesamerelativemolecularmassasthattheproteinexpectedandcouldberecognizedbyboththeanti-TRAILpolyclonalantibodyandanti-Hismonoclonalantibody.Inaddition,thisproteincouldalsoinhibitprolif-erationofhumanlymphomacelllineJurkatcellsorinduceapoptosisofthiscellline.ItisapparentthatarecombinantsolubleTRAILproteinwithbiologicalactivityisobtainedandthisprospectivestudycanlaysolidfoundationforfurtherresearchonthebiologicalactivityandapplicationintheanti-tumortherapy.

简介：Toinvestigatetheprotectiveeffectofepigallocatechingallate(EGCG)ontheimmunefunc-tionofdendriticcells(DCs)afterultravioletBirradiation(UVB)anditsunderlyingmechanisms,themonocyteswereisolatedfromperipheralbloodandcultivatedintoDCswithcytokines,suchasGM-CSFandIL-4.DCswereharvestedaftercultivationfor7dandsubjectedtoirradiationwithdifferentdosagesofUVB.Then,200μg/mlofEGCGwereaddedincertaingroupsimmediatelyafterirradiation.DCssimplytreatedwithUVBortreatedwithbothUVBandEGCGwereco-culturedwithlymphocytes,andMTFassaywasusedtodetecttheabilityofDCstostimulateproliferationoflymphocytes.SurfacemarkersCDS0,CD86,HLA-DRandCD40weredetectedbyflowcytometry,andthelevelsofIL-10andIL-12secretedfromDCs24haftercultivationweremeasuredbyELISA.ItwasdemonstratedthatUVBirradiationcouldinhibittheabilityofDCstostimulatetheproliferationoflymphocytesandsurfaceexpressionsofCDS0,CD86,HLA-DRandCD4onDCsinadose-dependentmanner.TheinhibitionrateofDCswasimprovedtosomeextentaftertreatmentwith200μg/mlofEGCG.Whentheconcentra-tionofEGCGexceeded100μg/ml,theenhancingeffectofEGCGontheexpressionoftheco-stimulat-ingmoleculesonDCscouldbedemonstratedinadose-dependentmanner.UVBshowednosignificantinfluenceonthesecretionofIL-10andIL-12fromDCs,whileEGCGcoulddown-regulatethesecretionlevelofIL-12andup-regulatethatofIL-10.ItisconcludedthatEGCGcanantagonizetheinhibitoryeffectonDCsinducedbyUVBirradiation.Thisfunctionhassomerelationshipwithitsprotectingeffectoftheexpressionoftheco-stimulatingmoleculeonthesurfaceofDCsandthesecretionlevelofIL-10andIL-12.

简介：AbstractToinvestigatetheroleofCD4^+helperT(Th)cellsinthememoryCTL-mediatedanti-tumorimmunity,theRAG-1geneknockoutmicewereadoptivelytransferredwithOT-1cellstogeneratethememoryCTL,theC57B1/6miceimmunizedwiththeepitopepeptideofOVAspecificThcellsandwithdifferentadjuvantswereadopfivelytransferredwiththesememory-CTLs,andthentheanimalswerechallengedwithtumorcellsEGT.ItwasfoundthatalthoughthesimpleimmunizationofmicewiththeepitopepeptideoftheOVAspecificThcellscouldgeneratemoreeffectCTL,butthiseffectwasnotsostrongenoughtoresistcompletelythechallengeswithtumorcells.Nevertheless,thememoryCTL-mediatedanti-tumorimmuneeffectrequiredthehelpsofTh1andTh2cells.Thecross-regulationbetweenThlandTh2cellsseemedtobebeneficialforthehosttogeneratemoreeffectorCTLformountinganefficientanti-tumorresponse.ItconcludedthattheinteractionbetweenThlandTh2cellsmightbemoreimportantthanthesinglesubsetofThcellsinthememoryCTL-mediatedanti-tumorimmuneresponse.Moreattentionshouldbepaidinthisregardforthefuturestudies.