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前列腺特异性膜抗原、粒细胞-巨噬细胞集落刺激因子双顺反子DNA疫苗的构建及其免疫活性的测定
黄云腾,叶传忠,陈方,齐隽
0
(上海交通大学医学院附属新华医院泌尿外科,上海,200092)
摘要:
目的:构建前列腺特异性膜抗原(PSMA)、粒细胞-巨噬细胞集落刺激因子(GM-CSF)核酸疫苗并测定其免疫活性.方法:应用DNA疫苗载体pIRES构建pIRES-PSMA-mGM-CSF、pIRES-PSMA、pIRES-mGM-CSF重组质粒,酶切鉴定正确后与pIRES空质粒分别免疫C56BL/6小鼠(每组15只),LDH释放试验测定各自免疫后小鼠的特异性细胞毒性T细胞(CTL)杀伤活性.结果:成功构建上述重组质粒;pIRES-PSMA-mGM-CSF免疫后小鼠特异杀伤率最高,pIRES-PSMA、pIRES-mGM-CSF次之,pIRES空质粒最差(P<0.05),各组杀伤效果以效靶比为101时最高.结论:PSMA及mGM-CSF双顺反子DNA疫苗有望在前列腺癌的基因治疗中发挥积极的作用.
关键词:  前列腺特异性膜抗原、粒细胞-巨噬细胞集落刺激因子、疫苗,DNA
DOI:10.3724/SP.J.1008.2006.00837
投稿时间:2006-07-04修订日期:2006-07-21
基金项目:
Construction of bicistronic DNA vaccine expressing prostate-specific membrane antigen and granulocyte-macrophage colony-stimulating factor and determination of its activity
HUANG Yun-teng,YE Chuan-zhong,CHEN Fang,QI Juan
(上海交通大学医学院附属新华医院泌尿外科,上海,200092)
Abstract:
Objective: To construct DNA vaccines expressing prostate-specific membrane antigen (PSMA) and/or granulocytemacrophage colony-stimulating factor (GM-CSF) and to determine their immunoactivity. Methods: Recombinant plasmids pIRES-PSMA-mGM-CSF , pIRES-PSMA, and pIRES-mGM-CSF were constructed with DNA vaccine vector plRES. Alter identified by endonuclease digestion, the above 3 plasmids and blank pIRES vector were used to immunize C56BL/6 mice (n= 15). LDH release assay was used to exam the cytotoxicity of cytolytic T lyrnphocytes in each group. Results: We successfully constructed the above mentioned recombinant plasmids. Mice in pIRES-PSMA-mGM-CSF immunized group had the highest specific cytotoxicity, followed by pIRES-PSMA and pIRES-mGM-CSF immunized groups. The blank pIRES group had the lowest cytotoxicity (P〈0.05). The cytotoxicity was the highest in all 4 groups at an effector/target ratio of 10/1. Condusion: The bicistronic DNA vaccine expressing PSMA and mGM-CSF may have a promising therapeutic value in gene therapy of prostate cancer.
Key words:  prostate specific membrane antigen  granulocyte-macrophage colony-stimulating factor  vaccine,DNA