mB7-1-GPI融合蛋白锚定Lewis细胞疫苗的制备及抗肿瘤作用

Preparation of mB7-1 -GPI anchored Lewis tumor cell vaccine and its anti-tumor effect

下载PDF阅读器目的 将mB7-1-GPI融合蛋白锚定于小鼠肺癌细胞株（Lewis）膜上,制备肿瘤细胞疫苗,研究该瘤苗的抗肿瘤作用.方法 将mB7-1-GPI融合蛋白锚定于小鼠肺癌细胞（Lewis）膜上,制备肿瘤细胞疫苗.采用流式细胞术检测该蛋白的细胞膜锚定作用.建立C57BL小鼠的肺癌模型,观察用mB7-1-GPI融合蛋白制备的肿瘤疫苗对荷瘤小鼠的免疫治疗作用.结果 mB7-1-GPI与Lewis瘤细胞共同孵育4 h后4℃放置0、4 h和8 h,样本的荧光强度分别为11.2、10.6和9.8,阳性细胞数百分率分别为95.8%、93.6%、91.1%.脾细胞＋Lewis组IL-2和IFN-γ分泌量分别为（25.9±1.4）pg/ml、（56.0±3.5）pg/ml,脾细胞＋Lewis/mB7-1-GPI组IL-2和IFN-γ分泌量分别为（871. 3±10.4）pg/ml和（1329.0±11.9）pg/ml,25 d时,Lewis/mB7-1-GPI瘤苗组小鼠的肿瘤直径（1.4±0.21）cm较Lewis瘤苗治疗组小（2.5±0.27）cm,差异有统计学意义（P＜0.05）.Lewis/mB7-1-GPI瘤苗组小鼠寿命为（75.2±2.0）d,与Lewis瘤苗组（40.2±1.3）d相比,生存期延长,差异有统计学意义（P＜0.05）.结论 mB7-1-GPI融合蛋白制备的肿瘤疫苗具有较强的抗肿瘤作用,有可能作为一种有效的新型疫苗用于肿瘤的治疗和预防.

Objective To prepare the mB7-1 -GPl-anchored Lewis vaccine and investigate its antitumor effects. Methods mB7-1 -GPI was incorporated on Lewis tumor cells and mB7-1 -GPI-anchoring tumor vaccine was prepared. The anti-tumor immunity induced by the prepared mB7-1 -GPI-anchored Lewis tumor cell vaccine in tumor-bearing mice was observed. Results Flow cytometric analysis showed that mB7-1 -GPI were positively expressed on the surface of Lewis tumor cells. After Lewis tumor cells incubated with mB7-1- GPI, the positive rate （PR） of mB7-1 antigen was 95.8% （Oh）, 93.6% （4h）, 91.1% （8h） and the fluorescence intensity （FI） was 11.2（0h） , 10.6（4h） , 9.8（8h）. The IL-2 and IFN-γ production of splenic lymphocytes ＋ lewis cells was （25.9 ± 1.4 ） pg/ml, （ 56. 0 ± 3.5 ） pg/ml. The IL-2 and IFN-γ production of splenic lymphocytes ＋ lewis/mB7-1-GPI was （ 871.3 ± 10. 4） pg/ml, （ 1329. 0 ± 11.9 ） pg/ml. In 25 days, the mean diameter of tumor of Lewis/mB7-1 -GPI was shorter than Lewis （ 1.4 ± 0. 21 ） cm ＆ （ 2. 5 ± 0. 27 ）em , P 〈 0. 05 ）. Lewis tumor cell-bearing C57BL/6 mice treated with Lewis/roBT.1 -GPI vaccine survived much longer than mice treated with Lewis vaccine （ 75.2± 2. 0 ） d ＆ （ 40. 2 ± 2. 0 ） d （ P 〈 0. 05 ）. Conclusion The Lewis tumor vaccine prepared with mB7-1 -GPI fusion protein significantly inhibited the tumor growth in Lewis beating mice. It represented an useful new strategy for attaching immunological factor onto tumor cell surfaces without genetic manipulation.