人β防御素-2与前列腺癌特异性膜抗原嵌合蛋白真核表达质粒构建及其诱导的小鼠特异性免疫应答  

Construction and Immunological Study of Recombinant hBD-2 /PSMA Chimeric Protein Eukaryotic Expressive Plasmid

作　　者：李明[1] 孙艳[1] 冯云[1] 吴琦[1] 黄宁[1] 王伯瑶[1] 

机构地区：[1]四川大学华西医学中心感染免疫研究室,成都610041

出　　处：《生物医学工程学杂志》2005年第2期283-287,共5页Journal of Biomedical Engineering

基　　金：CMB基金项目资助 (98-681)

摘　　要：探索人β防御素2 (h BD- 2 )和前列腺特异性膜抗原(PSMA)共表达重组核酸疫苗针对前列腺癌的免疫治疗。研究以pc DNA3.1为载体,构建重组质粒pc DNA3.1/PSMA和pc DNA3.1/h BD- 2 - PSMA,通过RT- PCR和免疫组化检测其表达。免疫小鼠后,进行血清中抗体检测,CD4 +、CD8+ T淋巴细胞数目测定及CTL 特异性杀伤作用检测。结果显示构建的质粒转染COS- 7细胞后能表达目的基因,免疫小鼠后能在体内持久表达,可以诱导产生特异性抗体,能有效的刺激T细胞增生,诱导特异性CTL 反应。当以h BD- 2作为免疫佐剂时,CTL 活性更强。本研究成功的构建了含PSMA的表达质粒,免疫小鼠可以诱导出有效的体液和细胞免疫。The recombinant PSMA DNA vaccine for active immunotherapy of prostate cancer was investigated. Two DNA vaccine recombinant plasmids, pcDNA3.1/PSMA and pcDNA3./hBD-2-PSMA, were constructed by inserting the hBD-2 gene and PSMA gene into an eukarytoic expression vector pcDNA3.1. Expression of the two recombinants was detected in transfected COS-7 cells immunohistochemical method. When immunized with immunized BALB/c mice acquired specific antibody and and inoculated mouse muscular cells by RT-PCR and pcDNA3.1/PSMA and pcDNA3./hBD-2-PSMA, the T cell response to PSMA. The quantity of the spleen lymphocytes and their CTL activity against PSMA gene transfected-BALB/3T3 cells significantly increased in the immunized mice, and the CTL activity of lymphocytes from pcDNA3.1/hBD-2-PSMA immunized mice was significantly higher than that of pcDNA3.1/PSMA immunized mice. This result suggests that pcDNA3.1/hBD-2-PSMA would probably be developed as a DNA vaccine for the immunotherapy of prostate cancer.

关 键 词：人Β防御素2 前列腺特异性膜抗原 核酸疫苗 免疫小鼠 前列腺癌 质粒构建 特异性免疫应答 防御素 真核表达 嵌合蛋白 

分 类 号：R737.25[医药卫生—肿瘤] R373[医药卫生—临床医学]