肝癌树突状细胞瘤苗诱导抗肿瘤作用的研究  

Antitumor effects induced by dendritic cell vaccine loaded with hepatocellular carcinoma antigen

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

机构地区：[1]广西医科大学附属肿瘤医院实验研究部,广西南宁530021

出　　处：《中华肿瘤防治杂志》2007年第11期805-808,共4页Chinese Journal of Cancer Prevention and Treatment

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

摘　　要：目的:研究负载肝癌抗原的肝癌树突状细胞(dendritic cell,DC)瘤苗诱导的抗肿瘤作用。方法:于体外用rhGM-CSF和rhIL-4从肝癌患者外周血诱导DC,并用肝癌细胞抗原冲击致敏,制成负载肝癌抗原的肝癌DC瘤苗。肝癌DC瘤苗活化自体T淋巴细胞分化为细胞毒性T淋巴细胞(cytotoxic of T-lymphocytes,CTL),采用流式细胞术检测CTL分型;乳酸脱氢酸(LDH)4 h释放法检测CTL对肝癌细胞的特异性杀伤作用。MTT法检测肝癌DC瘤苗及其活化CTL培养上清对肿瘤细胞的抑制作用;ELISA法检测肝癌DC瘤苗活化的CTL培养上清中IL-12、IFN-γ和TNF-α水平。结果:经肝癌DC瘤苗活化后的T淋巴细胞群中,CD56+细胞数量显著减少,CD4+T和CD8+T细胞数量增加,其中以CD8+T细胞增加较明显;细胞毒实验显示,该CTL对自体肝癌细胞具有较强的杀伤作用,而对与致敏DC的肝癌抗原无关的CT26结肠腺癌细胞无明显杀伤作用;单纯肝癌DC瘤苗或其激活的CTL培养上清也表现出较强的抑瘤作用,而且这种抑瘤作用具有广谱性,可抑制多种肿瘤细胞的生长;在CTL培养上清中可检测到IL-12、TNF-α和IFN-γ的含量。结论:肝癌DC瘤苗不仅诱导了对肝癌细胞的特异性CTL杀伤作用,而且可激活CD4+T和CD8+T细胞分泌IL-12、TNF-α和IFN-γ等细胞因子,以非杀伤方式直接或间接地抑制肿瘤细胞生长,发挥非特异性的抗肿瘤作用。OBJECTIVE： To estimate the antitumor responses induced by dendritic cell （DC） vaccine loaded with hepatocellular carcinoma（HCC） antigen. METHODS： DC isolated from the peripheral blood of HCC patients was induced with rhGM-CSF and rhIL-4 in vitro, then pulsed with HCC antigen to prepare DC vaccine loaded with HCC antigen. The subsets of tumor specific cytotoxic of T-lymphocytes （CTL） activated by DC vaccine loaded with HCC antigen were assayed by flow cytometry; the killing activity of CTL to HCC cells was tested by the lactate dehydrogenase （LDH） release assay in vitro, and the nonspecific inhibition of DC vaccine loaded with HCC antigen or the CTL culture supernatant against tumor cells was also detected. The levels of IL-12, TNF-α and IFN-γ in CTL culture supernatant were detected also by ELISA. RESULTS： After being stimulating by DC vaccine loaded with HCC antigen, the number of CD56^＋ cells decreased, however, the numbers of CD4^＋ T lymphocytes and CD8^＋ T lymphocytes both increased significantly, especially, the number of CD8^＋ T lymphocytes increased more significant. CTL could selectively kill HCC cells, but slightly kill CT26 tumor cells . The DC vaccine loaded with HCC antigen and the CTL culture supernatant could significantly inhibit the growth of tumor cells. The CTL culture supernatant had high levels of IL-12, TNF-α, IFN-γ. CONCLUSION.. The DC vaccine loaded with HCC antigen not only induce special CTL antitumor effects, but also induce non-special anti-tumor effects through activating the CD4^＋ T and CD8^＋ T cells to secrete IL-12 ,TNF-α and IFN-γ.

关 键 词：癌 肝细胞/免疫学 癌症疫苗 肝肿瘤 实验性/免疫学 树突细胞 

分 类 号：R735.7[医药卫生—肿瘤]