Ⅲ。．勰骂訾磊慧z⋯吼t叩，：170～174[文章编号]1007-385X(2003)03-0170-05肿瘤抗原负载的树突状细胞对肝癌肿瘤浸润淋巴细胞体外抗瘤作用的影响汪晓莺1，梁志强1，张一心2，孙伟红2，石佑琴1(I．南通医学院免疫学教研室，江苏南通226001；2．南通医学院附属医院外科，江苏南通226001)[摘要】目的：研究肿瘤抗原负载的树突状细胞(DCs)对原发性肝癌肿瘤浸润淋巴细胞(TILs)体外增殖和抗瘤作用的影响。方法：将自体肿瘤细胞匀浆粗提物(Tuly)与原发性肝癌患者外周血来源的DCs共培养以制备肿瘤抗原负载的DC(DC—Tuly)，将DC．Tuly与自体TILs在低浓度IL-2(100U／rnl)中共培养，用FCM测定DC—Tuly的表型以及与DC．Tuly混合培养前后的TIk表型，并计数TILs。用LDH法测定TILs的细胞毒作用。ELISA法检测ⅡLs培养上清中IFN一1和TNF—oL的含量。结果：DC。Tuly表面CDIa，CD83，CD80，CD86，CD40和HLA—DR分子表达水平增高(P<0．05)；DC—Tuly明显诱导TILs的增殖(P<0．01)及对自体肝癌细胞的细胞毒作用(P<0．01)，CD3+TILs增多(P<0．05)，CD4+TILs比例较混合前升高(P<0．01)，但仍以CD8+TILs为主。与DC—Tuly混合培养后第1，2天，7nk分泌IFN一1和TNF—d的量明显提高(P<0．01)。结论：负载肿瘤抗原的DCs可促进TILs的增殖，增强TILs的特异性抗肿瘤活性。[关键词]树突状细胞；肿瘤浸润淋巴细胞；细胞毒作用；原发性肝癌[中图分类号]R392．1；R735．7[文献标识码]AEffectofDendriticCellsLoadedwithTumorAntigensonAntitumorActivityofTumor-InfiltratingLymphocytesfromHepatocellularCarcinomainvitroWANGXiao—rin91，LIANGZhi—qian91，ZHANGK．xin2，SUNWei—hon91，SHIYou—qinl(J．DepartmentofIm—munologyofNantongMedicalCollege，Nantong226001；2．DepartmentofSurgeryofAffiliatedHospitalofNantongMedicalCollege，Nantong226001)[Abstract]0bjective：Toinvestigatetheeffectsofdendriticcells(DCs)loadedwithtumorantigensontheproliferationandantitumoractivityoftumor—infiltratinglymphocytes(TILs)fromhepatocellularcarcinoma(HCC)invitro．Methods：DCsderivedfromtheperipheralbloodofpatientswithHCCwereloadedwithautologoustumorlysate(Tuly)togenerateDC-TulyandthentheDC—TulywasCO—culturedwithautologousTILsinlOWconcentrationofIL．2．TILswerecounted．andthephenotypesofDCsandTILswereassayedbyFCMandthecytotoxicityofTILswasdeterminedwithLDHmethod．TheconcentrationofcytokinesinTILsculturesupernatantsuchasIFN一1andTNF．dwasdetectedbyELISA．Results：TheexpressionlevelsofCDla，CD80，CD83，CD86，CD40，HLA．DRmoleculesontheDC．Tulyarenotablyincreased(P<0．05)．TheproliferationofTILswhichwereinducedbyDC—Tulywasmarkedlyenhanced(P<0．01)andthecytotoxicity0ftheTILsagainstautologoustumorcellswasremarkablyincreased(P<0．01)．AfterCO．culturedwithDC．Tuly．CD3+TILswereincreased(P<0．05)andthepercentageofCD4+TILswasenhanced(P<0．01)，however，thepercentageofCD8+TILswasstillhigherthanthatofCD4+TILs．BothconcentrationsofIFN—YandTNF．0【intheculturesupernatantoftheTILsdistinctlyincreasedonthe1stand2nddaysofCO．culturing(尸<0．01)．Conclusion：DCsloadedwithtumorantigenscanobviouslypromotetheproliferationandthespecificantitumoractivityofTILs．[Keywords]dendriticcell；tumor—infiltratinglymphocyte；cytotoxicity；hepatocellularcarcinoma肿瘤浸润淋巴细胞(TILs)作为过继免疫治疗的效应细胞具有很高的抗瘤活性，以往人们用高浓度IL-2(1000～2000U／m1)体外诱导可使TIL明显扩增，但[基金项目]江苏省卫生厅资助课题(H9923)[作者简介】汪晓莺(1960一)，女，南通人。副教授，硕士，主要从事抗肿瘤、抗感染免疫研究万方数据中国肿瘤生物治疗杂志2003Sep；10(3)‘171’TIL对自体肿瘤的杀伤的特异性下降，而用低浓度IL-2(100U／m1)诱导，TIL却生长迟缓或停滞⋯。近年来发现，经肿瘤抗原致敏的树突...
