糖基化修饰的DC疫苗特异性抗骨髓瘤活性的体外研究  

In vitro investigation on specific anti-myeloma activity of the glycoengineering modified dendritic cell vaccine

作　　者：熊红[1] 吴秋业[2] 廖洪利[2] 赵庆杰[2] 侯健[1] 

机构地区：[1]第二军医大学长征医院血液科,上海200003 [2]第二军医大学药学院,上海200003

出　　处：《中国免疫学杂志》2007年第7期629-633,共5页Chinese Journal of Immunology

基　　金：上海市博士后科研资助计划项目(04R214134)

摘　　要：目的:探讨经过糖基化修饰改造过的肿瘤相关糖抗原冲击树突状细胞(DC)制备的DC疫苗特异性抗骨髓瘤作用。方法:采用化学方法及肿瘤细胞的生物工程法使骨髓瘤细胞表达新肿瘤相关抗原N-丙酰多聚唾液酸(NPrPSA);在无血清培养条件下,采用GM-CSF/IFN-α及TNF-α从MM患者外周血单核细胞诱导培养DC,然后用表达新抗原的肿瘤细胞冲击DC制备DC疫苗,将其刺激T细胞后,通过LDH释放法观察其对骨髓瘤细胞的杀伤效力,并采用ELISA法测定了T细胞分泌细胞因子IFN-γ的能力。结果:糖基化修饰的CD138+Npr-DC组LDH释放率较CD138+未修饰DC组明显增高(P<0.01)。CD138+Npr-DC组T细胞分泌IFN-γ与CD138+未修饰DC组相比亦明显增加(P<0.05),而CD138-Npr-DC组与CD138-未修饰DC组之间无统计学意义(P>0.05)。结论:糖基化修饰的DC疫苗可以有效地激活Th1细胞免疫应答,分泌高水平的细胞因子IFN-γ;可诱导出较普通DC疫苗更明显的抗骨髓瘤作用,并具有一定的安全性。Objective：To explore the specific anti-myeloma effect of the dendritic cell vaccine pulsed with tumor associated carbohydrate antigen modified by glycoengineering. Methods：With chemical and biological engineering methods, the new antigen, N- propionyl polysialic acid（ NprPSA）, was expressed on myeloma cells. Monocytes were cultured with serum-free culture medium containing GM-CSF/IFN-α and TNF-α to develop DC, which was pulsed with myeloma cells expressing the novel antigen to develop DC vaccine. After the autologous T cells were stimulated with the vaccine, the capability for killing the myelomt cells was observed by LDH release. And the IFN-γsecreted from T cells was detected by ELISA. Results：The LDH release in the modified CD138^＋ Npr-DC group was significantly higher than that in the CD138^＋ unmodified DC group（P 〈 0. 01 ）. The secretion of IFN-γ by the primed T cells was enhanced remarkably in CD138^＋ Npr-DC group in comparison with CD138^＋ unmodified DC （ P 〈 0.05 ）, but the difference between CD138^- Npr-DC and CD138^- unmodified DC group was no statistical significance（P 〉 0. 05 ）. Conclusion：The modified DC vaccine can activate efficiently Thl immunological response with high secretion of cytokine IFN-γ; and can induce stronger anti-myeloma effect than common DC vaccine, and has a certain safety.

关 键 词：多发性骨髓瘤 树突状细胞 疫苗 肿瘤相关糖抗原 

分 类 号：R733.3[医药卫生—肿瘤]