【摘要】 目的 :研究树突状细胞 (DC)的体外培养扩增及诱导特异性的抗肿瘤免疫反应。方法 :使用mGM CSF加mIL 4培养诱导骨髓细胞分化 ,采用反复冻融法制备L72 12白血病细胞的冻融抗原 (TAA) ,在DC培养的第3天加入TAA ,将TAA冲击致敏的DC与T淋巴细胞共培养 ,获得肿瘤特异性细胞毒性T淋巴细胞 (CTL) ,采用MTT法检测CTL对L72 12细胞的杀伤作用及其对野生型L72 12细胞再攻击的免疫保护作用。结果 :MTT检测发现CTL对L72 12细胞有特异性杀伤抑制作用 ,L72 12抗原冲击致敏的DC能显著提高和延长野生型L72 12细胞再攻击小鼠的生存率和存活期。结论 :mGM CSF和mIL 4配伍可有效地从小鼠骨髓细胞中诱导出大量的成熟的功能性DC ,L72 12白血病TAA冲击致敏的DC可诱导机体产生较强的抗肿瘤免疫反应更多还原

【Abstract】 Objective:To investigate specific antitumor immune response induced by dendritic cells (DC) from precursor in mice marrow.Method:The bone marrow cells were induced using granulocyte macrophage colony stimulating factor (mGM CSF) and/or interleukin 4 (mIL 4), respectively (①mGM CSF+ mIL 4,②mGM CSF,③ mIL 4,④ control). Morphological changes of DC were observed through electronic microscopy. Cells were harvested after 7 days and detected for phenotypic analysis by FACS. RT PCR method was used to determine the expression of CD80 in mature DC. After culture in 15% FCS RPMI1640 medium containing mGM CSF and mIL 4 for 3 days, DC were pulsed with L7212 tumor cytolysis antigen at the immature stage. The mature DC with special tumor antigen were collected on 7th day. After coculture for 3 days, the mature DC that were pulsed with L7212 tumor cytolysis antigen could activate T cells to become tumor specific cytotoxic T lymphocytes (CTL). Methyl thiazolyl tetrazolium (MTT) assay was employed to evaluate inhibition rate of CTL on L7212 cells and NIH3T3 cells, respectively. The increase of protective effect for the mice vaccinated with DC was observed.Result: DC precursors in mice marrow could be induced to differentiate and mature in medium containing mGM CSF and mIL 4 for 7 days. Mature DC with tumor cytolysis antigen could active naive T cells to become tumor specialized CTL. All the doses of CTL had significant inhibition or killing response on L7212 cells,but DC and inactivated T lymphocyte had no cytotoxicity on L7212 cells. After the DC vaccination, the survival time of the mice challenged with wild type L7212 cells were prolonged.Conclusion:In certain cytokine environment, the precursors in mice marrow can be induced to functional DC, which can activate naive T lymphocyte to CTL. Antitumor immune response could be induced by vaccination with antigen pulsed DC.更多还原