中文摘要：

目的观察CpG寡聚脱氧核苷酸（CpG-ODN）对截短的人前列腺特异性膜抗原（tPSMA）基因修饰的树突状细胞（DCs）诱导的抗前列腺癌效应。方法利用复制缺陷性腺病毒AdEasy-1系统，通过基因重组技术构建Ad—tPSMA及Ad—eGFP。将Ad-tPSMA和Ad．eGFP感染鼠源性DCs，用含10μg／L粒细胞-巨噬细胞集落刺激因子（GM-CSF）、白细胞介素（IL）-4和含10％胎牛血清（FCS）的RPMI1640培养基诱导培养6d后，然后再添加1mg／L的CpG—ODN体外培养1d，最后添加1mg／L的脂多糖（LPS）继续培养1d，使其成熟。流式细胞仪检测DCs细胞表型，CCK-8法检测混合淋巴细胞反应T细胞增殖能力，酶联免疫吸附试验（ELISA）试剂盒检测T细胞分泌细胞因子[IL-2、干扰素（INF）-γ]的影响，CCK-8试剂检测T细胞特性杀伤靶细胞活性。结果CpG．ODN促进Ad—tPSMA感染的DCs（CpG／DCs-Ad—tPSMA）高表达MHCII（83．8±3．7）％、CD80（79．8±5．6）％和CD86（78．3±2．8）％，且刺激同种异体T细胞增殖能力明显高于DCs—Ad—tPSMA组、DCs—Ad—eGFP组和未转染的DCs组（P〈0．05）；培养上清中细胞因子IL-2（179．64±2．72）ng／L和INF．1（1581．75±28．61）ng／L，表达水平均明显高于DCs—Ad—tPSMA组、DCs．Ad．eGFP组和未转染的DCs组（P〈0．05）；CpG／DCs—Ad—tPSMA组诱导RM-1-tPSMA细胞特异性杀伤率为（55．3±1．2）％，明显高于DCs—Ad—tPSMA组（40．7±1．4）％、DCs—Ad—eGFP组（12．7±1．2）％和未转染的DCs组（10．8±1．7）％（P〈0．05）。结论CpG—ODN能增强PSMA基因修饰的DCs疫苗诱导的特异性抗前列腺癌效应。

英文摘要：

Objective To observe cytosine-phosphorothioate-guanine oligodeoxynucleotides （CpG- ODN） for the anti-prostate cancer effect induced by dendritic cells （DCs） transduced with recombinant adenovirus vector bearing truncated human prostate specific membrane antigen （tPSMA） gene. Methods Replication deficient adenovirus AdEasy-1 system was used to construct recombination adenovirus Ad-tPSMA and Ad-eGFP. Mouse derived DCs were transduced with Ad-tPSMA and Ad-eGFP, and cultured for 6 Jays in the presence of 10 p.g/L GM-CSF and IL-4 in RPMI 1640 containing 10% FCS. After culture with 1 mg/L CpG- ODN for 1 day, DCs were further matured with lipopolysaccharide （LPS） at a concentration of 1 p.g/L for 1 day. The phenotype of DCs was analyzed by using flow cytometry. T cells proliferation stimulated by DCs in allogeneie mixed lymphocyte reactions and the level of interleukin （IL）-2, interferon （IFN）-γ,/were detected by ELISA kit, and cytotoxic CTL activity induced by DCs was tested by CCK-8 assay. Results CpG-ODN up-regulated the expression of MHCH （83. 8 ±3.7）%, CDSO （79. 8 ±5.6）% and CD86 （78. 3 ±2. 8）% in Ad-tPSMA-trandueed DCs （CpG/DCs-Ad-tPSMA）. T cells proliferation stimulated by CpG/DCs-Ad-tPSMA was significantly higher than that in DCs control group, DCs-Ad-tPSMA group and DCs-Ad-eGFP group （P 〈0. 05）. CpG/DCs-Ad-tPSMA induced increased IL-2 （179. 64 ±2. 72） ng/L, and INF-γ （ 1581.75 ± 28. 61 ） ng/L expression levels as compared to DCs control group, DCs-Ad-tPSMA group, and DCs-Ad-eGFP group （P 〈 0. 05 ）, and induced more outstanding RM-1-tPSMA cell specific cytotoxic rate （40. 7 ± 1.4 ）% than DCs control group （10. 8 ± 1.7）%, DCs-Ad-tPSMA group （40. 7 ± 1.4）%, and DCs-Ad-eGFP group （12. 7 ± 1.2）% （P 〈 0. 05）. Conclusion CpG-ODN can promote specific anti-prostate cancer induced by DCs modified with recombinant adenovirus vector bearing tPSMA gene.