肝癌患者与正常人外周血树突状细胞的诱导与功能的比较研究  

Functional comparison of induced dendritic cells from peripheral blood of patients with hepatocellular carcinoma and healthy individuals

作　　者：罗小玲[1] 谢裕安[1] 梁安民[1] 匡志鹏[1] 吴继宁[1] 

机构地区：[1]广西医科大学附属肿瘤医院实验研究部,南宁530021

出　　处：《中国癌症防治杂志》2012年第2期131-136,共6页CHINESE JOURNAL OF ONCOLOGY PREVENTION AND TREATMENT

基　　金：国家自然科学基金资助项目(30360114);广西自然科学基金资助项目(0448069)

摘　　要：目的对比研究肝癌患者和正常人外周血树突状细胞(DC)的诱导及其功能异同,探讨能否从肝癌患者外周血诱导出功能正常、具有抗肿瘤作用的DC疫苗。方法取肝癌患者和正常人外周血分离DC前体细胞,应用rhGM-CSF、rhIL-4及肝癌细胞抗原诱导DC。观察DC生长过程的形态学变化,以流式细胞术检测DC表面分子;ELISA法检测DC培养上清中IL-12水平;3H-TdR掺入法检测肝癌细胞抗原致敏DC刺激T细胞增殖的能力;乳酸脱氢酶4小时释放法检测肝癌细胞抗原致敏DC诱导的细胞毒性T淋巴细胞(CTL)的杀瘤活性。结果从肝癌患者和正常人外周血DC前体细胞诱导的DC具有相似的生长过程和细胞形态及功能,其表面分子HLA-DR、CD1a、CD83、CD80、CD86、ICAM-1的表达水平、DC培养上清中IL-12水平、DC刺激T细胞增殖的能力以及所诱生的CTL杀瘤活性等方面比较,差异均无统计学意义(P>0.05)。结论可从肝癌患者外周血诱导出功能正常并具有抗肿瘤作用的DC瘤苗,为以DC基础的肝癌的免疫治疗提供实验依据。Objective To investigate the potent anti-tumor activity of induced dendritic cells （DC）derived from the peripheral blood of patients with hepatocellular carcinoma（HCC）,and to lay the foundation for a DC vaccine against HCC. Methods DC were isolated from the peripheral blood of HCC patients and healthy individuals and induced in vitro using rhGM-CSF,rhlL-4 and HCC antigen. The morphology of DC was observed by phase contrast microscopy, and surface molecule expression was analyzed by fluorescence-activated cell sorting.IL-12 released by DC into the culture medium was detected by ELISA.T cell proliferation was quantitated by 3H- TdR uptake,and cytotoxic of T-lymphocyte （CTL）activity was assessed using the LDH release assay. Results Comparison of induced DC from HCC patients and induced DC from healthy individuals revealed no significant differences in growth characteristics ;expres- sion levels of surface molecules such as HLA-DR,CD1a,CD83,CD80,CD86,ICAM-1,and IL-12;and ability to stimulate T lymphocytes and induce CTL activity（P〉0.05 ）. Conclusions Induced DC from HCC patients can exert anti-tumor effects similar to those observed with induced DC from healthy individuals.These results provide an experimental basis for developing HCC immunotherapies involving DC.

关 键 词：肝肿瘤 树突状细胞 诱导 抗肿瘤作用 

分 类 号：R735.7[医药卫生—肿瘤]