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为了研制有效的疫苗 ,用于HPV16型重度感染和与其感染相关的宫颈癌晚期病人手术后的免疫治疗 ,用复制型DAN疫苗载体 pSCA1,在其CMVIE启动子之下插入修饰的HPV16型E7基因mE7- 3( 2 4G、2 6G、6 7R) ,构建成 pSCA1mE7- 3复制型DNA疫苗。以重组质粒免疫C5 7BL/ 6小鼠 ,检测诱发的特异性CTL活性 ;将免疫后的小鼠用TC - 1肿瘤细胞攻击 ,观察免疫保护效果。实验结果显示 :pSCA1mE7- 3复制型DNA疫苗能诱导小鼠产生针对TC - 1肿瘤细胞的特异性CTL反应 ;复制型DNA疫苗免疫后的小鼠能耐受 1× 10 4 TC - 1细胞的攻击 ,成瘤时间推迟 ,并且成瘤率明显下降 ,部分小鼠得到保护能免受肿瘤攻击。因此pSCA1mE7- 3复制型DNA疫苗可作为HPV16相关肿瘤的癌前病变及中晚期病人术后免疫治疗的候选疫苗。
In order to develop a potent vaccine for the immunotherapy of post-operative patients with advanced cervical cancer associated with HPV16-type severe infection and its infection, the replicative DNA vaccine vector pSCA1 was used to insert a modified HPV16 E7 gene, mE7, under its CMVIE promoter - 3 (2 4G, 2 6G, 6 7R) to construct pSCA1mE7-3 replicative DNA vaccine. The recombinant plasmid was used to immunize C5 7BL / 6 mice to detect the specific CTL activity. The immunized mice were challenged with TC - 1 tumor cells to observe the protective effect. The experimental results showed that the pSCA1mE7 - 3 replicative DNA vaccine induced specific CTL responses in TC - 1 tumor cells. The mice immunized with the replicative DNA vaccine were able to tolerate 1 × 10 4 TC - 1 cells , Tumor formation time delayed, and tumor formation rate decreased significantly, some mice are protected from tumor attack. Therefore, pSCA1mE7-3 replicative DNA vaccine can be used as a candidate vaccine for pre-cancerous lesion of HPV16-related tumors and post-operative immunotherapy of advanced patients.