rAAV/CEA转染树突状细胞制备肺癌疫苗  

Development of a lung cancer vaccine by transfecting dendritic cells with rAAV/CEA

作　　者：尤长宣[1] 钱晓涛[1] 何圆[1] Yong Liu Paul L Hermonat 

机构地区：[1]南方医科大学南方医院肿瘤科,广东广州510515 [2]美国阿肯色大学医学院基因治疗中心,小石城72205

出　　处：《南方医科大学学报》2014年第4期487-491,共5页Journal of Southern Medical University

基　　金：国家自然科学基金(81071847);广东省科技计划项目(2012B031800394);广东省自然科学基金(S2011010003881);吴阶平医学基金会临床科研项(320.6799.1118;320.6750.13288);南方医院高层次课题匹配项目~~

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(Dendritic cell,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]-TdR掺入法检测DC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-γ、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:⑧疫苗具备较强的刺激淋巴细胞增殖的能力,⑧诱导的CTL较高表达CD8、CD69和IFN-γ,⑧对CEA阳性的肺癌靶细胞产生较强杀伤活性,且此活性具有抗原特异性和MHC-I类限制性。结论以AAV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。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.

关 键 词：癌胚抗原 腺相关病毒 肺癌 疫苗 

分 类 号：R392[医药卫生—免疫学]