基因修饰树突状细胞诱导前列腺癌患者外周血T细胞亚群重排的研究

目的探讨以腺相关病毒（AAV）为载体,前列腺特异性抗原（PSA）基因转染树突状细胞（DC）诱导前列腺癌患者外周血T细胞亚群变化特点及临床意义。方法抽取30例前列腺癌患者外周血,采用密度梯度离心法分离外周血单个核细胞,以rAAV/PSA感染DC前体细胞,采用系列细胞因子诱导DC前体细胞成熟。第6天收集成熟DC并与T细胞按比例混合培养,诱导细胞毒性T淋巴细胞（CTL）。分别于DC与T细胞混合培养前后应用流式细胞术分析外周血T细胞亚群及调节性T细胞（CD4^＋CD25^＋FoxP3^＋Treg）的表达水平。结果 PSA基因转染DC刺激T淋巴细胞爆发增殖,与培养前比较,混合培养6d后CD8^＋、CD8^＋CD69^＋、CD8^＋CD28^＋T细胞的比例和CD8^＋/CD4^＋比值均明显增高,差异有统计学意义（P〈0.01）;而CD8＋CD28-T细胞和Treg细胞的比例均显著降低,差异有统计学意义（P〈0.01）。CD4^＋T细胞比例较前略有升高,但差异无统计学意义（P〉0.05）。结论 PSA基因转染DC能够有效地激活CD8＋抗原特异性CTL,下调免疫抑制性T细胞,提高患者的细胞免疫功能,为前列腺癌的免疫治疗提供新的有效策略。

Gene-transducted dendritic cells induce T lymphocyte subsets repopulation in peripheral blood of patients with prostate cancer

Objective To investigate the characteristics of rAAV/PSA-transducted dendritic cells（DCs） to induce T lymphocyte subsets repopulation in peripheral blood of patients with prostate cancer,and its clinical significance.Methods 30 patients with prostate cancer were included in this study.Peripheral blood mononuclear cells（PBMC,the DC precursor cells and lymphocytes） were isolated from prostate cancer patients by density gradient centrifugation and infected with rAAV/PSA virus.Maturation of the DC precursor was induced by series of cytokine.On day 6,the DCs were collected and mixed with T cells to induce cytotoxic T lymphocytes（CTL）.The proportion of T lymphocyte subsets and regulatory T cells（CD4^＋ CD25^＋ FoxP3^＋ Treg） in peripheral blood were analyzed by flow cytometry before and after mixed culture.Results The DCs transducted with PSA gene stimulated high proliferation of T cell populations.After mixed culture for 6 days,the percentage of CD8^＋,CD8^＋CD69^＋,CD8^＋ CD28^＋ T cells,the CD8^＋：CD4^＋ ratio were significantly increased（P〈0.01）,while the percentages of CD8＋CD28-T cells and Treg cells were markedly decreased（P〈0.01）.The percentages of CD4^＋ T cells of these two groups showed no significant difference（P〈0.05）.Conclusion The rAAV/PSA loading of DCs can effectively activate the CD8^＋ antigen-specific CTL,decrease immunosuppressive T cell and improve the cellular immunological function in patients.These findings may provide a potent immune therapeutic strategy for prostate cancer patients.