THERAPEUTIC EFFECT OF DENDRITIC CELLS TRANSDUCED WITH TH1-TYPE-CYTOKINE-GENES BY RECOMBINANT ADENOVIRUS VECTOR ON GASTROINTESTINAL TUMOR GROWTH

重组腺病毒载体转TH1型细胞因子基因的树突状细胞对胃肠道肿瘤生长的治疗作用

• 批准号: 12671198
• 负责人: GUNJI Yoshio
• 金额: $ 2.5万
• 依托单位: CHIBA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12671198/
• 关键词: DENDRITIC CELLS; ADENORICUS VECTOR; TH1 TYPE CYTOKINES; COLON 26; アデノウイルスベクター; Colon 26; Th1サイトカイン; IL-2

We have demonstrated that retrovirally transduced Th-1 type cytokine genes (Interleukin 2, 12, 15, 18 ) not Th-2 type cytokine genes( IL-4, IL-6) into colon.26 tumor cells elicit anti-tumor-effect. Growth of colon 26 /Th-1 type cytokine genes were successfully suppressed in syngeneic mice in our laboratory. In that experimental system, combined s.c. injection of colon 26/GM-CSF and /IL-4 tumor cells also elicit anti-tumor effect using potential activity of dendritic cells (DC)., Stimulation of the anti-tumor immune response by DC is critical to initiate anti-tumor immune machinery. To enhance this effect in tumor-bearing mice, bone marrow (BM) derived DC cells from Ba1b/C mice were genetically engineered to produce IL-2 by ex-vivo infection with a recombinant defective adenovirus (AxCAhIL-2). DC cells were generated from bone marrow cells cultured with mouse IL-4 (1000 u/ml) and GM-CSF(1000 u/ml) for 5 through 7 days and infected with AxCAhIL-2 with MOI 30. Those adenovirally transduced IL-2 gene produced high amount of IL-2 protein from DCs after infection. Intratumoral injection of IL-2 expressing DCs suppressed tumor growth profoundly, and prolonged the survival of the mice treated with those DCs (p<0.001). Multiple injection of DCs/AxCAhIL-2 enhanced the anti-tumor effect and increased the infiltration of macrophage to the peripheral site of tumor. These data suggest the possibility of DC/adenovirus mediated cytokine gene therapy.

我们已经证明，逆转录病毒将 Th-1 型细胞因子基因（白细胞介素 2、12、15、18）而非 Th-2 型细胞因子基因（IL-4、IL-6）转导到结肠 26 肿瘤细胞中，可引发抗肿瘤作用。我们实验室在同基因小鼠中成功抑制了结肠26/Th-1型细胞因子基因的生长。在该实验系统中，结合了 s.c.注射结肠26/GM-CSF和/IL-4肿瘤细胞也利用树突状细胞(DC)的潜在活性引发抗肿瘤作用。DC刺激抗肿瘤免疫反应对于启动抗肿瘤免疫机制至关重要。为了增强荷瘤小鼠的这种作用，对来自 Ba1b/C 小鼠的骨髓 (BM) 来源的 DC 细胞进行了基因改造，通过用重组缺陷腺病毒 (AxCAhIL-2) 进行离体感染来产生 IL-2。 DC细胞由与小鼠IL-4（1000 u/ml）和GM-CSF（1000 u/ml）培养5至7天的骨髓细胞产生，并用MOI 30的AxCAhIL-2感染。那些腺病毒转导的IL-2基因在感染后从DC中产生大量IL-2蛋白。瘤内注射表达 IL-2 的 DC 可显着抑制肿瘤生长，并延长用这些 DC 治疗的小鼠的存活时间 (p<0.001)。多次注射DCs/AxCAhIL-2增强了抗肿瘤作用并增加了巨噬细胞向肿瘤周围部位的浸润。这些数据表明DC/腺病毒介导的细胞因子基因治疗的可能性。

项目成果

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Shiiada, H 等人：“食管癌基因治疗”Res。

Matsubara, H et al: "Electroporation-mediated transfer of cytokine genes into human esophageal tumors produced anti-tumor effects in mice"Anticancer Research. 21. 2501-2504 (2001)

Matsubara, H 等人：“电穿孔介导的细胞因子基因转移到人食管肿瘤中，在小鼠中产生了抗肿瘤作用”抗癌研究。

Matsubara, et al.: "Electroporation-mediated transfer of cytokine genes into human esophageal tumors produced anti-tumor effects in mice"Anticancer Research. 21. 2501-2504 (2001)

Matsubara 等人：“电穿孔介导的细胞因子基因转移到人食管肿瘤中，在小鼠中产生了抗肿瘤作用”抗癌研究。

Matsubara, H, Gunji Y, et al.: "Electorporation-mediated transfer of cytokine genes into human esophageal tumors produced anti-tumor effects in mice"Anticancer Research. 21. 2501-2504 (2001)

Matsubara, H, Gunji Y 等人：“电穿孔介导的细胞因子基因转移到人食管肿瘤中，在小鼠中产生抗肿瘤作用”抗癌研究。

Yoshio Gunji: "Antitumor effect of bone marrow-derived dendritic cells engineered to produce interleukin-2 by adenoviral vector."Molecular therapy. 1. S269-S270 (2000)

Yoshio Gunji：“通过腺病毒载体改造骨髓源性树突状细胞产生白细胞介素 2 的抗肿瘤作用。”分子治疗。