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1、·244·中国免疫学杂志2004年第20卷肺癌细胞总RNA转染的DC疫苗体外诱导肺癌患者①特异性抗肿瘤免疫的实验研究任秀宝 李 于津浦 刘 虹 郝希山 (天津医科大学附属肿瘤医院免疫室,天津300060) 中国图书分类号 R39219 文献标识码 A 文章编号 10002484X(2004)0420244205[摘 要] 目的:为了探讨肺癌细胞总RNA转染的DC疫苗体外诱导特异性抗肿瘤免疫的能力。方法:采用分离肺癌患者外周血单核细胞体外诱导DC细胞,Trizol法提取肺癌细胞系Calu2
2、6总RNA,用脂质体包裹总RNA转染DC并诱导特异性CTL的扩增,LDH法和ELISA法检测CTL的杀伤活性和IFN2γ分泌。结果:经肺癌细胞总RNA转染的DC特异性表面标志及功能相关分子表达均上调,转染后的DC可显著刺激异体P自体T淋巴细胞增殖,诱导的特异性CTL对携带Calu26抗原的靶细胞的杀伤率显著高于LAK细胞,再次接触相同抗原时其IFN2γ分泌量显著增高。结论:肺癌细胞总RNA转染的DC疫苗可在体外诱导出特异性抗肿瘤免疫。[关键词]树突状细胞;肺癌;RNA;肿瘤免疫;疫苗Experim
3、entalstudyofspecificantitumorimmunityinducedbydendriticcellvaccinetransfectedwithtumorcelltotalRNAinlungcancerpatientsRENXiu2Bao,LIZheng,YUJin2Pu,LIUHong,HAOXi2Shan.ImmunologyDepartment,TianjinCancerInstituteandHospital,Tianjin300060,China[Abstract]Ob
4、jective:ToobserveabilityofDCvaccinetransfectedwithtumorcelltotalRNAtoinducespecificantitumorimmunityinlungcancerpatientsinvitro.Methods:DCsweregeneratedfromlungcancerpatients’peripheralbloodmononuclearcells(PBMC).TotalRNAwasisolatedfromlungcancertumor
5、celllineCalu26byTrizol.AutologousDCstransfectedwithCalu26totalRNAbyliposomewereusedtoin2ducespecificCTLproliferation.SpecificcytotoxicityandIFN2γsecretionweremeasuredbyLDHassayandELISAmethod.Results:TransfectedDCsexhibiteddramaticallyincreasedexpressi
6、onofspecificmembranemarkersandfunction2associatedmolecules,andweremorepotentinstim2ulatingallogenousorautologousTcellproliferationthanthatofcontrolDCs.SpecificCTLsinducedbytransfectedDCsshowedhighercytotoxici2tythanLAKagainstCalu26antigenspositivetarg
7、etcells.WhensensitizedlymphocyteswererestimulatedbytransfectedDCsagain,IFN2γsecre2tionenhancedsignificantly.Conclusion:Lungcancerpatient’sautologousDCstransfectedwithtumortotalRNAareeffectivevaccinesinstimu2latingspecificantitumorT2cellimmunityinvitro
8、.[Keywords]Dendriticcell;Lungcancer;RNA;Antitumorimmunity;Vaccine 肺癌,尤其是非小细胞肺癌术后对于放疗和化1 材料与方法疗的敏感性较差,应用生物治疗方法是近几年来异军突起的重要辅助治疗手段。树突状细胞(Dendritic111 试剂和细胞系 RPMI1640完全培养基、DM2cells,DC)是人体内功能最强大的抗原呈递细胞,对RIE2C脂质体、TricolReagent均购自GIBCOBRL,鼠启动初次免疫应答有