TGF-β不敏感前列腺癌特异性CTLs的制备及其抗癌作用

Preparation of TGF-β insensitive cytotoxic T lymphocyte against prostate cancer and its anti-tumor activity

目的:制备TGF-β不敏感前列腺癌特异性CTLs,观察其对前列腺癌的治疗作用。方法:分离前列腺癌患者外周血单个核细胞,体外诱导DCs;制备该患者前列腺癌肿瘤抗原,冲击DCs后诱导肿瘤特异性CTLs;用携带TβRⅡDNglytk基因的慢病毒感染肿瘤特异性CTLs;Western blotting检测TβRⅡDNglytk感染后CTLs对TGF-β的敏感性。分别以TGF-β敏感和不敏感CTLs治疗小鼠前列腺癌移植瘤,比较其疗效。结果:前列腺癌组织抗原冲击后DCs诱导的肿瘤特异性CTLs生长速度快于未冲击DCs组(P<0.01),其活化状态标志CD25的表达显著高于对照CTLs(P<0.01)。TβRⅡDNglytk慢病毒感染使肿瘤特异性CTLs对TGF-β敏感性减弱。TGF-β不敏感的CTLs可显著抑制荷瘤鼠前列腺癌的生长(P<0.01)。结论:成功制备的TGF-β不敏感前列腺癌特异性CTLs对小鼠前列腺移植瘤有明显的治疗作用,为前列腺癌免疫治疗奠定了一定的实验基础。

Objective：To prepare TGF-β insensitive cytotoxic T lymphocytes（CTLs）against prostate cancer and to observe its therapeutic effect against prostate cancer. Methods： Peripheral blood mononuclear cells were obtained from prostate cancer patients, and were induced to differentiate into dendritic cells in vitro. Prostate cancer antigen was prepared from the same patients, and tumor antigen specific CTLs were induced by prostate tumor antigen impulsedDCs. The tumor antigen specific CTLs were infected with lentivirus containing TβRⅡDNglytk gene, and the response of TβRⅡDNglytk transfectedCTLs to TGFβ was examined by Western blotting analysis. Prostate cancer cellinoculated mice were treated with TGF-β sensitive and insensitiveCTLs separately, and the therapeutic results were compared. Results： CTLs induced by prostate tumor antigen impulsedDCs grew significantly faster than those induced by nonimpulsedDCs （P〈001）. TβRⅡDNglytk gene infection reduced the sensitivity of CTLs to TGFβ. The reaction of CTLs infected by lentiviral vector to TGF-β was significantly weaker than that of noninfected CTLs. TGF-β insensitive CTLs significantly inhibited the growth of implanted prostate tumors in mice. Conclusion： TGF-β insensitive prostate tumor antigen specific CTLs have apparent therapeutic effect against prostate cancer, which paves a way for immunotherapy of prostate cancer.