负载CD133+肝癌细胞RNA树突状细胞疫苗的免疫活性  

Immunological Competence of Dendritic Cell Vaccine Loaded with CD133+ Hepatocellular Carcinoma Cell RNA

作　　者：易平[1] 张志明[1] 林家耀 任远[1] 欧阳高雄 刘春辉[1] 冯钟煦[1] 吕庆杰[1] 刘剑勇[1] 

机构地区：[1]广西医科大学附属肿瘤医院肝胆外科,南宁530021

出　　处：《肿瘤防治研究》2017年第3期184-188,共5页Cancer Research on Prevention and Treatment

基　　金：广西科学研究与技术开发计划攻关项目(桂科攻1355005-3-3)

摘　　要：目的探讨负载人肝细胞性肝癌(HCC)组织来源的CD133+肝癌细胞RNA树突状细胞(CD133+HCC RNA-DC)疫苗的免疫活性。方法采用酶消化法从人HCC组织中分离出肝癌细胞,利用流式细胞术分选出CD133+肝癌细胞,制备负载CD133+肝癌细胞RNA树突状细胞疫苗,最后用流式细胞术检测DC的表型,ELISA法测定DC分泌IL-12水平,采用混合淋巴细胞反应法检测DC在体外刺激自体淋巴细胞增殖的能力。结果 CD133+肝癌细胞RNA树突状细胞的HLA-ABC、HLA-DR、CD86、CD80、CD83表达水平分别是(96.52±2.02)%、(92.17±3.04)%、(94.25±3.28)%、(55.14±1.67)%、(40.53±2.31)%,与肝癌细胞RNA树突状细胞和成熟DC比较,差异均无统计学意义(P>0.05)。CD133+肝癌细胞RNA-DC、肝癌细胞RNA-DC、成熟DC和未成熟DC分泌IL-12的量分别为(421.50±3.12)、(418.20±1.10)、(324.20±2.19)和(102.47±4.60)pg/ml,前两者之间差异无统计学意义(P=0.14),前两者均高于后两者,差异有统计学意义(P<0.05)。CD133+肝癌细胞RNA-DC与肝癌细胞RNA-DC刺激自体T淋巴细胞增殖能力分别均强于成熟DC和无DC刺激的自体T淋巴细胞,差异均有统计学意义(P<0.05)。结论CD133+肝癌细胞RNA树突状细胞疫苗具有成熟表型,能够在体外有效刺激自体T淋巴细胞增殖。Objective To investigate the immunological competence of dendritic cell loaded with CD133＋ hepatocellular carcinoma cell RNA（CD133＋ HCC RNA-DC） vaccine.Methods HCC cells were separated from hepatocellular carcinoma tissues through Enzyme Digestion,and then CD133＋ HCC cells were sorted by flow cytometry.CD133＋ HCC RNA-DC vaccine was obtained.Flow cytometry was used to detect the phenotype of DC,and ELISA was applied to determine the level of DC＇s secretion of IL-12.Mixed lymphocyte reaction was applied to test the ability of DC to stimulate the proliferation of autologous T lymphocytes in vitro.Results The expression levels of HLA-ABC,HLA-DR,CD86,CD80 and CD83 in CD133＋ HCC RNA-DC group were（96.52±2.02）%,（92.17±3.04）%,（94.25±3.28）%,（55.14±1.67）% and（40.53±2.31）%,respectively.Phenotypes expression levels were not significantly different among CD133＋ HCC RNA-DC,HCC-DC and mature DC.The secretion of IL-12 in CD133＋ HCC RNA-DC group,HCCDC group,mature DC group and immature DC group were（421.50±3.12）,（418.20±1.10）,（324.20±2.19） and（102.47±4.60）pg/ml,respectively;the difference of the former two had no statistical significance（P=0.14）,and the amounts of the former two were higher than that of the latter two（P〈0.05）.The proliferation of autologous T lymphocytes stimulated by CD133＋ HCC RNA-DC and HCC-DC were stronger than those by mature DC and no DC（P〈0.05）.Conclusion The dendritic cell vaccine loaded with CD133＋ hepatocellular carcinoma cell RNA（CD133＋ HCC RNA-DC） has mature phenotype and can effectively stimulate the proliferation of autologous T lymphocytes in vitro.

关 键 词：CD133 肝细胞性肝癌 RNA 树突状细胞疫苗 

分 类 号：R735.7[医药卫生—肿瘤]