基于TEM-8新型颗粒性疫苗的制备、鉴定和体外激发CTL杀伤活性的研究

Preparation and identification of a novel particle vaccine based on pTEM-8 and its ability to stimulate CTL cytotoxic activity

目的制备、鉴定基于肿瘤抗原TEM-8(tumor endothelial marker-8)的复合颗粒性疫苗,研究其在体外激发CTL杀伤活性。方法在特定的条件下将阳离子肽和DNA制备成复合颗粒性的结构,以透射电镜、DNA阻滞试验、DNa-seI保护试验、Westernblot等对该疫苗进行制备和鉴定,同时进行体外激发CTL杀伤活性的标准51Cr释放实验。结果在NaCl浓度为87.5mmol/L、电荷比为2的制备条件下,电镜扫描结果显示该疫苗能形成颗粒;Westernblot鉴定其可有效介导TEM-8蛋白表达;该疫苗有明显的杀伤毒性,杀伤率为59.7%,与对照组性比较,差异有统计学意义(P<0.05)。结论成功制备了基于肿瘤抗原TEM-8的复合颗粒性疫苗,其在体外能够有效激发出特异性CTL应答。

Objective To prepare and identify the particles tumor vaccine based on tumor endothelial marker-8 （TEM-8） and stimulated cytotoxic activity of CTL in vitro. Methods Under specific condition, peptide and DNA were condensed into particle, and the preparing parameters were optimized by assays of DNA gel retardation, DNase I protection, and identified by Western blotting and 51Cr release assay. Results The vaccine was condensed into homogeneous particles at charge ratio of 2 in 87.5 mmol/L NaCl concentration, which effectively medicated the expression of the genes carried by vaccine particles. Conclusion The tumor vaccine based on pTEM-8 is prepared successfully, and induces specific CTL with specific cytotoxic effect. These results provide a good foundation for the design of new tumor vaccine.