摘要
目的制备一种能高效激活免疫细胞的肿瘤疫苗,提高免疫细胞杀伤肿瘤细胞的功能。方法同源重组表达含人免疫球蛋白（Ig G）Fc片段的MUC1蛋白作为抗原,并利用DOTAP阳离子脂质体包被抗原呈递给体外诱导培养的树突状细胞（DC）,然后用DC细胞激活产生特异性免疫应答的T细胞,检测T细胞对肿瘤细胞的杀伤能力。结果成功构建含人免疫球蛋白（Ig G）Fc片段的MUC1蛋白表达载体,经大肠杆菌成功表达目的蛋白并纯化,经聚丙烯酰胺凝胶电泳验证得到目的蛋白条带。抗原蛋白经DOTAP阳离子脂质体包被,包封率达到97%。包被后的抗原刺激体外培养的免疫细胞,流式细胞术检测发现1~40μg/ml抗原制剂均能有效激活产生免疫应答,并显著增加功能性肿瘤杀伤细胞亚群比例,联合对肿瘤细胞（MCF-7）杀伤功能分析实验,10μg/ml疫苗制剂效果最佳,能提高免疫细胞对肿瘤细胞杀伤率45%以上。结论制备的疫苗制剂能有效被DC细胞捕获,并激活产生特异性应答的功能性肿瘤杀伤细胞,显著提高了免疫细胞对肿瘤细胞的杀伤功能。
The study aimed design a tumor therapeutic vaccine targeting MUC1 which can efficiently activate immune cells and improve the function of immune cells against tumor cells. Through homologous recombination,MUC1 protein with human immunoglobulin（Ig G） Fc fragment as an antigen was expressed, and coated with DOTAP cationic liposomes, then presented to dendritic cells（DC）. The DC cells were used to activate specific immune response of T cells, then the ability of kill tumor cells of the T cells was tested by CCK-8 assay. Date showed that MUC1-Fc protein expression vector was constructed successfully, and the protein was expressed by E. coli and characterized and confirmed by polyacrylamide gel electrophoresis. The antigen protein was successfully coated by DOTAP cationic liposomes and the entrapment rate was 97%. In vitro, the coated antigen can effectively activate immune response and significantly increase the subsets of the anti-tumor functionality cells. With 10 μg/ml vaccine, the anti-tumor（against MCF-7） rate was enhanced more than 45%. In conclusion, a vaccine that significantly improves the function of T cells against tumor is successfully constructed.
出处
《免疫学杂志》
CAS
CSCD
北大核心
2015年第11期964-968,共5页
Immunological Journal
基金
深圳市海外高层次人才创新创业专项资金(KQCX20120806161031208)
