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中国癌症防治杂志

中国癌症防治杂志 ›› 2012, Vol. 4 ›› Issue (2): 131-135.doi: 10.3969/j.issn.1674-5671.2012.02.09

• 肝癌专栏 • 上一篇    下一篇

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

  

  1. 广西医科大学附属肿瘤医院实验研究部
  • 出版日期:2012-06-25 发布日期:2012-07-03
  • 通讯作者: 罗小玲 E-mail:lxllc001@sina.com.cn
  • 基金资助:

    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）

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

  • Online:2012-06-25 Published:2012-07-03

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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。

关键词: 肝肿瘤, 树突状细胞, 诱导, 抗肿瘤作用

Abstract: 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，rhIL-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；expression 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.

Key words: Liver neoplasms, Dendritic cell, Induction, Anti-tumor effect