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[目的]探讨CB6F1小鼠脾树突状细胞(DC)的培养及其诱导针对小鼠路易斯肺癌(LLC)的细胞毒性T淋巴细胞(CTL)对肿瘤的杀伤效应。[方法]应用CB6F1小鼠脾细胞在GM-CSF、IL-4等细胞因子作用下培养出DC,反复冻融法制备LLC抗原致敏DC,与淋巴细胞及IL-2混合培养诱导出肿瘤特异性CTL,利用乳酸脱氢酶法检测CTL的杀伤活性。[结果]用GM-CSF、IL-4联合培养小鼠脾细胞第4d,可见细胞形态发生改变,培养第8d,可见典型的刺突样DC,通过流式细胞术检测了DC表型高表达CD80占76.5%,CD86占60.0%,MHCⅡ占67.4%,CD11C占80.6%。[结论]应用GM-CSF、IL-4、LPS等细胞因子培养CB6F1小鼠脾细胞经过肿瘤抗原冲击,可以培养出成熟DC,并且DC可以诱导出具有杀伤活性的肿瘤特异性CTL。
[Objective] To investigate the culture of splenic dendritic cells (DCs) of CB6F1 mice and the cytotoxicity induced by cytotoxic T lymphocytes (CTLs) against mouse Lewis lung carcinoma (LLC). [Method] The CB6F1 mouse spleen cells were cultured under the action of cytokines such as GM-CSF and IL-4 to produce DCs. LLC antigen-primed DCs were prepared by repeated freeze-thaw cycles and mixed with lymphocytes and IL-2 to induce tumor-specific CTL CTL was detected by lactate dehydrogenase assay. [Results] The results showed that the morphology of cells was changed on the 4th day after spleen cells were cultured with GM-CSF and IL-4. On the 8th day after culturing, typical spike-like DCs were observed. The DC phenotypes were highly expressed by flow cytometry CD80 accounted for 76.5%, CD86 accounted for 60.0%, MHC Ⅱ accounted for 67.4%, CD11C accounted for 80.6%. [Conclusion] Mature DCs can be induced from spleen cells of CB6F1 mice after being cultured with cytokines such as GM-CSF, IL-4 and LPS, and tumor-specific CTLs with cytotoxicity can be induced by DCs.