CpG+ODN对人外周血树突状细胞分化、成熟与抗肿瘤作用影响的实验分析

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1、英文摘要TheExperimentalStudyofEffectofCpGODNOnDifferentiation、MaturationandAntitumorofHumanPeripheralBloodDendriticCellsABSTRACTGraduateStudent：LIU"fanTutor：YANGNingBasicalSchoolofMedicalScienceobjective：ToexploretheeffectsOtCpGODNoffthedlfferentiation、maturationanda

2、ntitumorofhumanperipheralblooddendriticcells．TbinvestigatetheregulationofCpGODNtoDCculturedwithtumorcellsupernatant。Methods：(1)Afierisolatedfromnormalhumanperipheralblood，DCisexamineditspuritybyS-100proteinimmunocytochemistrystaining．(2)Thefactorialdesignisapplie

3、dtoanalyzetheeffectofdifferentlevelsofCpGoDNconcerntration、DCconcerntrationandculturetimeonDCgrowth，andthebestconditionofCpGODN-stimulatedDCinvitroisselected．(3)ThecellmorphologyofDCthatincubatedwithandwithoutCpGODNareobservedbytransmissonelectronmicroscope。theex

4、pressionofsurfaceantigenHLA-DR、CD86onthemisanalyzedbyflowcytometry(FCM)andtheeffectofthemontheproliferationofnaiveTcellinallogeneicmixedlymphocytereaction(MLR)andantitumorexperimentaredetectedbyMTTtest．(4)Thesupernatantoftumorcellarecollectedandusedasthecondition

5、aIculturemediumforDCfreshlyisolatedfromhumanperipheralblood．MTTcalorimetricassayisusedtodetecttheeffectofdifIbrentconcemtrationoftumorcelIsupematantonthegrowthofDCatdifferenttime，andtheexpressionofHLA—DRonDCculturedwithandwithoutsupernatantisanalyzedbyFCM．(5)DCis

6、olatedfreshlyisjncubatedwithlO％FCSRPMll640culturemedium，CpGODN(1um01．r)，tumorcolIsupematant(30％)andCpGODN(1um01．1。)+tumorcellsupematant(30％)for72h，respectively．TheDCarecharacterizedbyproliferativeandphenotypicanalysis．Results：CI)S-100proteinimmunocytochemistrysta

7、ininganalysisshowsthatDCspurity’90％(2)ThefactorialtestexplainsthatCpGODNconcerntrationhassignificanteffectonthegrowthofDC(p<0．01)．AIso，withDCconcemtratlonhasasynergisticeffectonit．ThebestconditionofCoGODN—stimulatedDC：CpOODNis1ut001．rI，DCis6x1矿well～andfor36h，(3)T

8、hemorphologicalandfonctionalalaysisofDCshowthatthesurfaceofCpGODN-stimulatedDChasmanythin、longandregulardendriteswithnumerousroughendoplasmicreticulumandscanty