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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 a composite particle vaccine based on tumor antigen marker TEM-8 (tumor endothelial marker-8) and study its cytotoxic activity against CTL in vitro. Methods The cationic peptides and DNA were prepared into composite granular structures under specific conditions. The vaccine was prepared and identified by transmission electron microscopy, DNA blockade assay, DNa-seI protection assay and Western blot. Meanwhile, CTLs Active standard 51Cr release assay. Results Under the condition of NaCl concentration of 87.5mmol / L and charge ratio of 2, the immunofluorescence showed that the vaccine could form granules. Western blotting could effectively induce the expression of TEM-8 protein. The vaccine had obvious cytotoxicity, The killing rate was 59.7%, compared with the control group, the difference was statistically significant (P <0.05). Conclusion The composite particle vaccine based on the tumor antigen TEM-8 was successfully prepared, which can effectively stimulate the specific CTL response in vitro.