DC与CIK共培养后对白血病细胞杀伤活性的研究[1].pdf

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1、实用癌症杂志2005年9月第20卷第5期　ThePracticalJournalofCancer,September2005,Vol20,No.5·465·DC与CIK共培养后对白血病细胞杀伤活性的研究李　曼　王　春　陈宝安　李翠萍　傅　强　　【摘要】　目的　研究细胞因子诱导的杀伤细胞(CIK)与同源树突状细胞(DC)共培养后CIK细胞的增殖活性、表型的变化,及其对K562、HL260白血病细胞细胞毒作用的影响。方法　采集健康供者的外周血单个核细胞(MNC),置于37℃,5%CO2培养箱培养2h,收集非贴壁细胞用于诱导培养CIK细胞,贴壁细胞诱导分化出成熟DC,将成熟D

2、C和CIK细胞按1’5的比例混合培养3天,用MTT法检测DC2CIK共培养细胞杀伤K562和HL260白血病细胞株的活性。结果　DC2CIK共培养后增殖速度明显快于++++单纯CIK细胞组(P<0.05);培养第14天,CIK中CD3CD8、CD3CD56双阳性细胞的比率分别为58.6%±7.3和26.5%±++++6.2,DC2CIK的CD3CD8、CD3CD56的比率分别为72.5%±4.2和38.4%±6.1,表达差异显著(P<0.05);在2.5∶1～20∶1的效靶比范围内,DC2CIK对K562、HL260细胞的杀伤活性较单纯CIK细胞组的杀伤活性要高(P<0

3、.05)。结论　DC与CIK共培养细胞是增殖活性和细胞毒活性均高于CIK细胞的免疫活性细胞。【关键词】　树突状细胞;细胞因子诱导的杀伤细胞;白血病;免疫治疗;K562细胞株;HL260细胞株中图分类号:R733.7　　文献标识码:A　　文章编号:100125930(2005)0520465204StudyoftheCytotoxicityEffectofCo2culturedDendriticCellsWithCytokineInducedKillerCellsonLeukemiaCellsLIMan,WANGChun,CHENBao2an,etal.TheAffili

4、atedGulouHospitalofNanjingUniversity,Nanjing,210008【Abstract】ObjectiveTostudythecytotoxicityeffectofco2cultureddendriticcells(DCs)withcytokineinducedkillercells(CIKs)onK562、HL260leukemiacelllines.MethodsPeripheralbloodmononuclearcells(PBMC)wereisolatedfromheathyadultdonors.After2hoursinc

5、ubationofPBMC,DCswereinducedfromadherentcellsbysomecytokinesandCIKsweregeneratedfromnon2adherentcells.MatureDCswereharvestedafter9daysincubation,andthenco2culturedwithCIKsataratioof1to5.Thecyto2toxicityactivityofco2culturedDC2CIKsagainstK562celllineandHL260celllinewasinvestigatedbyMTTass

6、ays.ResultsCom2paredwithCIKs,theco2culturedDC2CIKspresentedamarkedlyhigherproliferation(P<0.05).On14thofincubation,thepositive++++ratesofCD3CD8、CD3CD56cellswere58.6%±7.3and26.5%±6.2inCIKs,and72.5%±4.2and38.4%±6.1inco2cul2turedDC2CIKs,showingasignificantdifferencebetweenthetwogroups(P<0.0

7、5).CIKswereabletolyseK562andHL260celllinesatloweffectortotargetratios.Thiseffectwassignificantlyenhancedbyco2culturingwithDCs(P<0.05).ConclusionCo2cul2turedDC2CIKsarehighlyeffectiveimmunecellswhichhasahigherproliferationandcytotoxityagainstK562andHL260celllinesthanCIKs.【K