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携带突变型K-ras基因的重组腺病毒转染树突状细胞诱导的体外抗肿瘤免疫反应

In vitro anti-tumor immune response induced by dendritic cells transfected with recombinant adenovirus carrying mutant K-ras genes.

作者信息

Zhao Feng, Zhou Qinghua, Lu Yanrong, Qin Yang, Zhang Jie, Li Jinsong, Wang Jianjun

机构信息

Department of Thoracic Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China.

出版信息

J Huazhong Univ Sci Technolog Med Sci. 2005;25(4):378-81. doi: 10.1007/BF02828201.

Abstract

The specific anti-tumor immune response induced by mouse bone marrow dendritic cells (DCs) transfected with recombinant adenovirus carrying mutant k-ras genes was investigated. DCs were generated from mouse bone marrow in the presence of rmGM-CSF (3.3 ng/mL) and rmIL-4 (1.3 ng/mL) and detected by FACS, and then transfected with the recombinant adenovirus encoding mutant k-ras gene. The efficacy of transfection and T cell stimulating activity of DCs were detected. CTL activity of the mice vaccinated with DCs was observed. The results showed that DCs had dendritic veiled morphology. BmDCs highly expressed B7-1 (80%), B7-2 (77%), MHC II (70%), CD11c (65%), CD40 (70%) and CD54 (96%) with FACS, and no significant difference in the expression was observed before and after the transfection (P > 0.05). The DCs transfected by mutant k-ras gene could significantly stimulate lymphocytes proliferation as compared with those transfected by Ad-c or non-modified DCs (P < 0.05). DC vaccine transfected by mutant k-ras gene could induce CTL activity against Lewis lung cancer, but not against B16. The specific cytotoxicity against Lewis lung cancer in Ad-k-ras/12-transduced DC group was significantly higher than those in the control, vector and non-transfected DCs groups (P < 0.05). It was concluded that special antitumor response could be induced by DCs transfected with recombinant adenovirus carrying mutant k-ras genes.

摘要

研究了用携带突变型k-ras基因的重组腺病毒转染的小鼠骨髓树突状细胞(DCs)诱导的特异性抗肿瘤免疫反应。在重组人粒细胞巨噬细胞集落刺激因子(rmGM-CSF,3.3 ng/mL)和重组人白细胞介素-4(rmIL-4,1.3 ng/mL)存在的情况下从小鼠骨髓中生成DCs,并通过流式细胞术进行检测,然后用编码突变型k-ras基因的重组腺病毒进行转染。检测了DCs的转染效率和T细胞刺激活性。观察了接种DCs的小鼠的细胞毒性T淋巴细胞(CTL)活性。结果显示DCs具有树突状面纱样形态。骨髓来源的DCs(BmDCs)通过流式细胞术检测显示高表达B7-1(80%)、B7-2(77%)、主要组织相容性复合体II类分子(MHC II,70%)、CD11c(65%)、CD40(70%)和CD54(96%),转染前后表达无显著差异(P>0.05)。与用腺病毒-c(Ad-c)转染的DCs或未修饰的DCs相比,用突变型k-ras基因转染的DCs能显著刺激淋巴细胞增殖(P<0.05)。用突变型k-ras基因转染的DC疫苗可诱导针对Lewis肺癌的CTL活性,但对B16无效。Ad-k-ras/12转导的DC组对Lewis肺癌的特异性细胞毒性显著高于对照组、载体组和未转染DCs组(P<0.05)。结论是携带突变型k-ras基因的重组腺病毒转染的DCs可诱导特异性抗肿瘤反应。

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