NEW DEVELOPMENT OF DC IMMUNOTHERAPY TO UNIDENTIFIED CANCER ANTIGEN

DC免疫治疗未知癌抗原的新进展

• 批准号: 15591330
• 负责人: TAKAYAMA Takuya
• 金额: $ 2.11万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15591330/
• 关键词: denderitic cell; in vivo electroporation; Flt3L; IL-18; chemokine; SLC; in vitro electroporation

1)IN VIVO MOBILIZATION AND MATURATION OF DENDRITIC CELLS USING FLT3L GENE THERAPYThe clinical application of DCs induced from peripheral monocytes in vitro has been initiated as a promising immunological therapy against cancer. If the same type of immuno-stimulator could be achieved without in vitro manipulation, it might be very convenient in clinical settings. In this study, we performed systemic gene transfer of Flt3L using in vivo electroporation of Flt3L plasmid DNA (Flt3L-IVE) in pretibial muscles in order to determine the effects on DCs in situ. The number of DCs was significantly increased and showed highly co-stimulatory molecules expressions both in spleen and bone marrow after Flt3L-IVE compared to those of control groups. Immunohistochemical evaluation revealed that not only DCs but also CD8 and CD4 positive cells were significantly infiltrated into the local tumor site compared with those of control and remained in the tumor 21 days after a single Flt3L-IVE. However, anti- … More tumor effects of Flt3L-IVE were not significant in MCA205 established tumor. These results suggest that Flt3L gene transfer using in vivo electroporation could mobilize DCs into tumor site. Additional means to induce maturation of these DCs could have positive impact on anti-tumor effects of this strategy.2)INDUCTION OF POTENT AND SYSTEMIC ANTITUMOR IMMUNITY USING INTRA-TUMORAL GENE TRANSFER OF IL-18 IN COMBINATION WITH FLT3L THERAPYInterleukin-18 (IL-18), which induces IFN-γ production and enhances the cytolytic activity of NK cells, is considered to be a good candidate to be used for cancer immunotherapy. However, we have already found that treatment with systemic or local administration of IL-18 alone was not enough to induce a potent systemic anti-tumor immunity. On the other hand, we also have reported that dendritic cells (DCs) can capture tumor antigens from tumor cells killed by NK cells activated with IL-18 and efficiently induce tumor-specific CTL in vitro. In order to enhance the systemic anti-tumor response induced by local administration of IL-18 in vivo, we examined the effects o」 the combination therapy with IL-18 and Flt3 ligand (Flt3L). The mice received intra-dermal inoculation of MCA205 fibrosarcoma in the bilateral flanks. On day 5 and 12, mice were treated with in vivo electroporation (IVE) with DNA plasmids carrying cDNA of human Flt3L or EGFP to bilateral hind legs. As the combination therapy, some of the mice were also treated with intra-tumoral injection of adenoviral vector carrying IL-18 gene (Ad.IL-18) or EGFP gene (Ad.EGFP). In the treated tumors, significant anti-tumor effect was observed in mice treated with Ad.IL-18 alone and the ones treated with combination therapy of Flt3L-IVE and Ad.IL-18 when compared to those mice with control (p<0.01). The combination treatment with Ad.IL-18 resulted in the more potent anti-tumor response when compared to Flt3L-IVE treatment alone (p<0.01), and the complete eradication was observed more frequently (100% vs 33% : p<0.05) in mice treated with the combination therapy when compared to ones with Ad.IL-18 treatment alone. In the un-injected tumors, only the combination therapy showed significant anti-tumor. Lymphoid cells in regional lymph nodes of the mice treated with the combination therapy showed a significant cytolytic activity against MCA205. Moreover, cytolytic activity of the combination therapy against YAC-1 (NK target) was significantly higher than that of Ad.IL-18 treatment alone (p<0.05). These results suggested that local gene transfer of IL-18 combined with DCs mobilization in situ with Flt3L may enhance the anti-tumor effect and induce a potent systemic anti-tumor immunity. Less

1）FLT 3L基因治疗对树突状细胞的体内动员和成熟外周血单核细胞体外诱导的树突状细胞作为一种有前途的抗肿瘤免疫治疗方法已开始临床应用。如果不需要体外操作就可以获得相同类型的免疫刺激剂，那么在临床环境中可能会非常方便。在这项研究中，我们进行了系统的Flt 3L基因转移，在体内电穿孔Flt 3L质粒DNA（Flt 3L-IVE）在胫前肌，以确定对原位DC的影响。与对照组相比，Flt 3L-IVE组小鼠脾脏和骨髓中的DC数量明显增加，并显示出高水平的共刺激分子表达。免疫组化结果显示，与对照组相比，Flt 3L-IVE诱导后21 d，肿瘤组织中不仅有DC，而且有CD 8和CD 4阳性细胞浸润，并在肿瘤组织中存在。然而，反- ...更多信息 Flt 3L-IVE的肿瘤效应在MCA 205建立的肿瘤中不显著。这些结果表明，使用体内电穿孔的Flt 3L基因转移可以动员DC进入肿瘤部位。诱导这些DC成熟的其他方法可能对该策略的抗肿瘤效果产生积极影响。2）使用IL-18的肿瘤内基因转移联合FLT 3L治疗诱导有效和免疫性抗肿瘤免疫白细胞介素-18（IL-18），其诱导IFN-γ产生并增强NK细胞的细胞溶解活性，被认为是用于癌症免疫治疗的良好候选者。然而，我们已经发现，单独全身或局部施用IL-18的治疗不足以诱导有效的全身抗肿瘤免疫。另一方面，我们也报道了树突状细胞（DCs）可以从IL-18激活的NK细胞杀伤的肿瘤细胞中捕获肿瘤抗原，并在体外有效地诱导肿瘤特异性CTL。为了增强由体内局部施用IL-18诱导的全身抗肿瘤应答，我们检查了用IL-18和Flt 3配体（Flt 3L）的组合疗法的效果。在小鼠的双侧胁部接受MCA 205纤维肉瘤的皮内接种。在第5天和第12天，用携带人Flt 3L或EGFP的cDNA的DNA质粒对小鼠双侧后腿进行体内电穿孔（IVE）处理。作为联合治疗，一些小鼠还通过瘤内注射携带IL-18基因（Ad.IL-18）或EGFP基因（Ad.EGFP）的腺病毒载体进行治疗。在治疗的肿瘤中，当与具有对照的那些小鼠相比时，在用Ad.IL-18单独治疗的小鼠和用Flt 3L-IVE和Ad.IL-18的组合疗法治疗的那些小鼠中观察到显著的抗肿瘤效果（p<0.01）。当与单独Flt 3L-IVE治疗相比时，与Ad.IL-18的组合治疗导致更有效的抗肿瘤应答（p<0.01），并且当与单独Ad.IL-18治疗相比时，在用组合治疗治疗的小鼠中更频繁地观察到完全根除（100%对33%：p<0.05）。在未注射的肿瘤中，只有联合治疗显示出显着的抗肿瘤作用。用组合疗法处理的小鼠的局部淋巴结中的类淋巴细胞显示出针对MCA 205的显著的细胞溶解活性。此外，针对YAC-1（NK靶标）的联合治疗的细胞溶解活性显著高于单独的Ad.IL-18治疗的细胞溶解活性（p<0.05）。这些结果表明，IL-18局部基因转移联合Flt 3L原位动员DC可增强抗肿瘤效应，并诱导有效的全身抗肿瘤免疫。少

项目成果

Generation of Mature Dendritic Cells Fully Capable of T Helper Type 1 Polarization Using OK-432 Combined with Prostaglandin E2

使用 OK-432 与前列腺素 E2 结合生成完全具有 T 辅助细胞 1 型极化能力的成熟树突状细胞

• 发表时间: 2003
• 期刊: Cancer Science 94
• 作者: Sato M;Takayama T;et al.
• 通讯作者: et al.

Takayama T, Kaneko K, Morelli AE, Li W, Tahara H, Thomson AW: "Retroviral delivery of transforming growth factor-beta1 to myeloid dendritic cells : inhibition of T-cell priming ability and influence on allograft survival."Transplantation. 74巻1号. 112-119 (

Takayama T、Kaneko K、Morelli AE、Li W、Tahara H、Thomson AW：“将转化生长因子-β1 逆转录病毒递送至骨髓树突状细胞：抑制 T 细胞启动能力并影响同种异体移植存活。” 1号。112-119（

Generation of Mature Dendritic Cells Fully Capable of T Helper Type 1 Polarization Using OK-432 Combined with Prostaglandin E2.

使用 OK-432 与前列腺素 E2 结合生成完全具有 1 型 T 辅助细胞极化能力的成熟树突状细胞。

• 发表时间: 2003
• 期刊: Cancer Sci 94
• 作者: Sato M;Takayama T;Tanaka H;Konishi J;Suzuki T;Kaiga T;Tahara H.
• 通讯作者: Tahara H.

Cancer gene therapy using in vivo electroporation of Flt3-ligand.

• DOI: 10.3892/ijo.27.2.457
• 发表时间: 2005-08
• 期刊: International journal of oncology
• 影响因子: 5.2
• 作者: Kazuya Shimao;T. Takayama;K. Enomoto;Tetsuya Saito;S. Nagai;J. Miyazaki;K. Ogawa;H. Tahara