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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,为前列腺癌的免疫治疗奠定了一定的实验基础
To explore the co-expression of human beta-defensin 2 (h BD-2) and prostate-specific membrane antigen (PSMA) recombinant nucleic acid vaccines for the immunotherapy of prostate cancer. The pc DNA3.1/PSMA and pc DNA3.1/h BD-2 - PSMA recombinant plasmids were constructed using pc DNA3.1 as a vector, and their expression was detected by RT-PCR and immunohistochemistry. After immunization of mice, serum antibodies, CD4 +, CD8 + T-lymphocyte counts and CTL-specific killing were detected. The results showed that the constructed plasmid could express the target gene after transfected into COS-7 cells. After being immunized, the plasmid could be expressed in vivo for a long time. It can induce the production of specific antibodies, which can effectively stimulate the proliferation of T cells and induce specific CTL responses. When hBD-2 was used as an immunoadjuvant, CTL activity was stronger. This study successfully constructed an expression plasmid containing PSMA. The immunized mice can induce effective humoral and cellular immunity, laying a certain experimental basis for the immunotherapy of prostate cancer.