中文摘要：

目的探讨以改构腺相关病毒2型（AAV-2）为载体，将rAAV/CEA转染树突状细胞（Dendriticcell，DC）制备肺癌治疗性疫苗的可行性。方法取健康人外周血，分离获得贴壁的单个核细胞（Mo），以rAAV/CEA病毒转染，对照组以CEA多肽刺激，两组细胞均以重组人粒细胞巨噬细胞集落刺激因子（GM-CSF）、白细胞介素4（IL-4）、肿瘤坏死因子-α（TNF-α）诱导DC成熟。第7天收集DC，取DC与原始T细胞混合，诱导细胞毒性T细胞（CTL），[^3H]-T诹掺入法检测刺激自体T细胞增殖能力；流式细胞仪分析CTL细胞中IL-4、IFN-y、CD8、CD4、CD69、CD25的表达情况；取CEA阳性的肺癌细胞株A549为靶细胞，采用51Cr释放法检测CTL对该靶细胞的杀伤效率。结果rAAV/CEA转染的DC疫苗可诱导特异性细胞免疫，具体表现在：（陂苗具备较强的刺激淋巴细胞增殖的能力，②诱导的CTL较高表达CD8、CD69和IFN-γ，③对CEA阳性的肺癌靶细胞产生较强杀伤潘眭，且此活性具有抗原特异性和MHC-I类限制性。结论以AAAV为载体，rAAV/CEA基因转染DC可成功制备针对肺癌的治疗性疫苗。

英文摘要：

Objective To study the feasibility of preparing a therapeutic lung cancer vaccine by transfecting dendritic cells （DCs） with adeno-associated virus vector carrying carcino-embryonic antigen gene （rAAV/CEA）. Methods Adherent cells （monocytes） isolated from the peripheral blood of a healthy donor were infected with rAAV/CEA virus stock or pulsed with CEA peptide （control）. The monocytes in both groups were induced into mature DCs with recombinant human GM-CSF, IL-4 and TNF-α. At day 7 of induction, the mature DCs were harvested and mixed with T lymphocytes. T cell proliferation stimulated by the DCs was assessed with 3H-thymidine uptake, and the expression of IL-4, IFN-7, CD8, CD4, CD25 and CD69 in cytotoxic T lymphocytes （CTL） was analyzed with flow cytometry. The cytotoxicity of the CTL against the target CEA-positive lung cancer A549 cells was tested by 51Cr releasing assay. Results The DCs transfected with rAAV/CEA strongly stimulated the proliferation of the T cell populations, and the induced CTL showed high expressions of CD8, CD69 and IFN-γ. The transfected DCs exhibited a high killing ability of CEA-positive lung cancer cells, and the killing showed a CEA antigen specificity and was limited by MHC I. These results suggested the ability of rAAV/CEA-transfected DCs in generating specific cellular immunity in vitro. Conclusion It is feasible to prepare therapeutic lung cancer vaccines by transfecting DCs with rAAV/ CEA.