负载gp96-ki67复合物的DC疫苗对骨肉瘤的抗肿瘤效应体外实验研究  

An in vitro study of the anti - tumor effects of DC vaccine loaded with gp96 -ki67 complexes on osteosarcoma cells

作　　者：刘林 丁利伟 严专 李慧忠 

机构地区：[1]徐州医科大学附属医院急救中心,江苏徐州221002 [2]江苏省肿瘤生物治疗研究所,江苏徐州221002

出　　处：《徐州医学院学报》2016年第11期701-704,共4页Acta Academiae Medicinae Xuzhou

摘　　要：目的负载gp96-ki67复合物的树突状细胞（DC）疫苗对骨肉瘤细胞系U2—OS的杀伤作用及其机制。方法用抗原肽复合物gp96-Ki67、gp96、Ki67、PBS分别负载DC，然后与同源的T淋巴细胞共培养7天，流式细胞仪检测各组T淋巴细胞分泌穿孔素情况；ELISA法检测各组释放γ-干扰素（INF-γ）浓度；LDH释放实验及CFSE／7AAD双标法检测体外对骨肉瘤细胞系U2—OS的杀伤效率。结果gp96-Ki67组T淋巴细胞分泌穿孔素、IFN-γ浓度最强，与gp96组、Ki67组、PBS组比较，差异均有统计学意义（P〈0．05）。各组T淋巴细胞对U2-OS细胞的杀伤率随着效靶比的增加而提高，在效靶比为40：1时，各组的杀伤率最高；gp96-Ki67组杀伤作用最强，与Ki67组、gp96组、PBS组比较，差异有统计学意义（P〈0．05）。结论gp96-Ki67复合物能很好地修饰DC，使DC有效地提呈抗原，提高T淋巴细胞分泌穿孔素和IFN-γ的浓度，在体外对骨肉瘤有很强的抗肿瘤作用。Objective To investigate the cytotoxicity of dendritic cells （DC） loaded with gp96 - ki67 peptide complex against osteosarcoma cell line U2 - OS and the mechanism involved. Methods Dendritic cells were loaded with gp96 - ki67 peptide complex, gp96, Ki67 and PBS respectively. Then, the cells were co - cultured with T lymphocytes for 7 days. The level of perforin secreted by T cells in each group was detected by flow cytometry. The amount of interfer- on -γ （INF- γ） released in each group was detected by ELISA. The cytotoxicity of Ki67 on U2 -OS cells was examined by LDH release assay and CFSE/7AAD double - labeled assay in vitro. Results The gp96 - ki67 group produced higher amounts of perforin and IFN -γ, compared with the gp96, Ki67 and PBS groups （P 〈 0.05 ）. As the ratio of ef- fector to target increased, the cytotoxicity of T cells against U2 - OS was enhanced and reached the peak when the ratio of effector to target was 40 ： 1. The gp96 - ki67 group presented the significantly strongest cyctotoxicity compared with the Ki67, gp96 and PBS groups. Conclusion Loaded with gp96 - ki67 complex, DC can effectively present antigen, increase the amounts of perforin and IFN - γ, and produce strong effects against osteosarcoma in vitro.

关 键 词：树突状细胞 Ki67抗原肽复合物 细胞毒性T淋巴细胞 骨肉瘤 

分 类 号：R738.7[医药卫生—肿瘤]