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[目的]研究荷载肿瘤干细胞(CSCs)抗原的树突状细胞疫苗联合白介素-2(IL-2)治疗小鼠恶性黑色素瘤的研究。[方法]利用Aldefluor染料从小鼠恶性黑色素瘤细胞系D5中分选出肿瘤干细胞,将CSCs制备成细胞裂解液加载在DC细胞上制备成CSCs-DC疫苗。将D5细胞接种至C57BL/6小鼠皮下建立小鼠恶性黑色素瘤模型,随机分成磷酸盐缓冲液组(PBS)、白介素-2组(IL-2)、CSCs-DC疫苗组(CSCs-DC)、CSCs-DC疫苗联合白介素-2组(CSCsDC+IL-2),并给予相应的治疗。测量并记录各组肿瘤大小变化和小鼠生存期,实验终点取小鼠脾脏,体外培养激活后利用ELISA法测定B细胞分泌Ig G水平,LDH细胞毒性实验测定细胞毒性淋巴细胞(CTL)活性。[结果]CSCs-DC+IL-2组肿瘤生长明显慢于PBS组,IL-2组及CSCs-DC组(P<0.05)。小鼠生存期CSCs-DC+IL-2组最长为58.33±1.67天,CSCs-DC组为51.67±0.33天,较IL-2组43.33±2.40天长,PBS组平均生存时间最短,为34.00±1.53天。CSCs-DC+IL-2组小鼠脾脏中B细胞分泌Ig G水平和CTL活性最高,CSCs-DC组次之,IL-2组较CSCs-DC组和CSCs-DC+IL-2组低,但较PBS组高,差异有统计学意义(P<0.01)。[结论]CSCs-DC疫苗联合IL-2方案在治疗恶性黑色素瘤的小鼠模型中效果较CSCs-DC疫苗组和IL-2组好,与IL-2提高T/B细胞活性有关。
[Objective] To study the treatment of mouse malignant melanoma with dendritic cell vaccine combined with interleukin-2 (IL-2) loaded with tumor stem cell (CSCs) antigen. [Method] The cancer stem cells were sorted out from mouse malignant melanoma cell line D5 by using Aldefluor dye. The CSCs were prepared into cell lysate and loaded on DC cells to prepare CSCs-DC vaccine. D5 cells were inoculated subcutaneously into C57BL / 6 mice to establish murine malignant melanoma model and randomly divided into PBS group, IL-2 group and CSCs-DC vaccine group (CSCs-DC) , CSCs-DC vaccine combined with interleukin-2 group (CSCsDC + IL-2), and given the appropriate treatment. The changes of tumor size and survival time of mice in each group were measured and recorded. The spleen of the mice was taken at the end of the experiment. The levels of Ig G secreted by B cells were measured by ELISA and the activity of cytotoxic lymphocytes (CTL) was measured by LDH cytotoxicity. [Results] The tumor growth in CSCs-DC + IL-2 group was significantly slower than that in PBS group, IL-2 group and CSCs-DC group (P <0.05). The CSCs-DC + IL-2 group had the longest survival time of 58.33 ± 1.67 days and the CSCs-DC group was 51.67 ± 0.33 days, which was 43.33 ± 2.40 days longer than that of IL-2 group. The shortest survival time was 34.00 ± 1.53 days. The CSCs-DC + IL-2 group had the highest level of Ig G secretion and CTL activity in the spleen of the mice, followed by the CSCs-DC group, the IL-2 group was lower than the CSCs-DC group and the CSCs-DC + But higher than PBS group, the difference was statistically significant (P <0.01). [Conclusion] CSCs-DC vaccine combined with IL-2 regimen is better than CSCs-DC vaccine group and IL-2 group in the treatment of malignant melanoma mouse model, which is related to the increase of T / B cell activity by IL-2.