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目的构建表达前列腺特异性膜抗原(PSMA)的DNA疫苗,观察其在体外对肿瘤细胞的免疫攻击和在体内对肿瘤细胞攻击的免疫保护作用。方法通过稳定转染构建表达PSMA的小鼠黑色素瘤细胞系B16-PSMA,将DNA疫苗pCDNA3.1-PSMA通过肌肉注射导入C57BL/6小鼠体内,分离小鼠脾细胞,检测细胞毒性T淋巴细胞(cytotoxicTlymphocytes,CTL)反应。以B16-PSMA细胞攻击免疫后小鼠,观察免疫动物的无瘤生存期和肿瘤体积增长情况,评价DNA疫苗的抗肿瘤作用。结果DNA疫苗可诱导小鼠脾淋巴细胞CTL活性,经过免疫后的小鼠成瘤率降低,无瘤生存期延长,肿瘤生长缓慢,肿瘤组织内有较多淋巴细胞浸润,表明产生较强的抗肿瘤反应。结论表达PSMA的DNA疫苗能够诱导小鼠产生特异性免疫反应,对表达PSMA的肿瘤细胞的攻击产生免疫保护作用,为前列腺癌的预防和免疫治疗提供了新的思路。
Objective To construct a DNA vaccine expressing prostate-specific membrane antigen (PSMA) and observe its immunological protection against tumor cells in vitro and the immune protection against tumor cells in vivo. Methods The murine melanoma cell line B16-PSMA expressing PSMA was constructed by stable transfection. The DNA vaccine pCDNA3.1-PSMA was intramuscularly injected into C57BL / 6 mice, and the splenocytes of mice were isolated and detected for cytotoxic T lymphocytes (cytotoxicTlymphocytes, CTL) reaction. The immunized mice were challenged with B16-PSMA cells to observe the tumor-free survival and tumor volume growth of the immunized animals, and to evaluate the anti-tumor effect of the DNA vaccine. Results DNA vaccine could induce CTL activity in splenic lymphocytes of mice. The immunized mice showed reduced tumor formation rate, prolonged tumor-free survival, slow tumor growth and more infiltration of lymphocytes in the tumor tissue, indicating stronger anti-tumor effect Tumor response. Conclusions PSMA-expressing DNA vaccine can induce specific immune responses in mice and protect the PSMA-expressing tumor cells from challenge. It provides a new idea for prophylaxis and immunotherapy of prostate cancer.