An experimental study on cancer vaccine therapy using dendritic cells genetically transduced with the tumor antigen gene and the ubiquitin gene

使用转染肿瘤抗原基因和泛素基因的树突状细胞进行癌症疫苗治疗的实验研究

• 批准号: 17591342
• 负责人: NAKAMURA Masaki
• 金额: $ 2.48万
• 依托单位: Wakayama Medical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-17591342/
• 关键词: Dendritic cells (DCs); CEA; Adnovirus vector; Ubiquitin; アデノウイルベクター

Cancer immunotherapy using dendritic cells (DCs) adenovirally transduced with the whole tumor-associated antigen (TAA) gene is an effective approach. In this study, we established the carcinoembryonic antigen (CEA) -specific cytotoxic T lymphocytes (CTLs) using an in vitro stimulation with adenovirally modified human DCs that express CEA. Moreover, DCs were transduced with the fusion gene of the CEA gene and the ubiquitin gene, and we showed the more potent cytotoxic activity against CEA-expressing targets (A24). Streptococcal preparation OK-432 is useful for stimulating DCs in terms of the maturation. To augment therapeutic antitumor effect, we investigated whether the OK-432 stimulation would be more effective on inducing CEA-specific CTLs compared with the other typical stimuli. DCs were cultured under various conditions, tumor necrosis factor (TNF) -a, lipopolysaccharide (LPS) or OK-432. A cytotoxic assay using peripheral blood mononuclear cells (PBMCs) -derived CTLs was performed in a 4h^<-51>Cr release assay. OK-432 stimulated immature DCs to acquire mature phenotype and to produce significant amounts of T-helper 1 cytokines. In all groups (immature DCs, TNF-a/DCs, LPS/DCs, OK-432/DCs), CEA-specific CTLs were generated. OK-432-stimulated DCs (HLA-A24) induced the most potent cytotoxic activity against CEA-expressing targets (A24), and not against controls. OK-432/DCs could induce markedly potent CTLs specific to the target cells pulsed with CEA652 peptide (HLA-A24-restricted peptide), although others failed to induce potent CTLs. In conclusion, the CTLs induction protocol using adenovirally modified DCs transduced with the fusion gene of the CEA gene and the ubiquitin gene or after maturation with OK-432 showed a potent antitumor activity against CEA-expressing target cells, and is therefore promising for clinical applications as a cancer vaccine therapy.

利用树突状细胞腺病毒转导全肿瘤相关抗原（TAA）基因进行肿瘤免疫治疗是一种有效的方法。在这项研究中，我们利用腺病毒修饰的表达CEA的人树突状细胞体外刺激建立了癌胚抗原（CEA）特异性细胞毒性T淋巴细胞（ctl）。此外，我们用CEA基因和泛素基因的融合基因转导DCs，并对表达CEA的靶标（A24）显示出更强的细胞毒活性。链球菌制剂OK-432在刺激dc成熟方面是有用的。为了增强抗肿瘤治疗效果，我们研究了与其他典型刺激相比，OK-432刺激是否更有效地诱导cea特异性ctl。在肿瘤坏死因子(TNF) -a、脂多糖（LPS）或OK-432等不同条件下培养DCs。使用外周血单个核细胞（PBMCs）衍生的ctl进行细胞毒性试验，在4h^<-51>Cr释放试验中进行。OK-432刺激未成熟的dc获得成熟表型，并产生大量的t -辅助性1细胞因子。在所有组（未成熟的DCs、TNF-a/DCs、LPS/DCs、OK-432/DCs）中，都产生了cea特异性的ctl。ok -432刺激的树突状细胞（HLA-A24）对表达cea的靶点（A24）具有最有效的细胞毒活性，而对对照没有作用。OK-432/ dc可以诱导具有CEA652肽（hla - a24限制性肽）脉冲的靶细胞特异性的明显有效的ctl，尽管其他细胞不能诱导有效的ctl。综上所述，用CEA基因和泛素基因融合基因转导的腺病毒修饰的dc或经OK-432成熟后诱导的ctl诱导方案对表达CEA的靶细胞具有有效的抗肿瘤活性，因此有望作为癌症疫苗治疗的临床应用。

项目成果

Tumor vaccine therapy against recrudescent tumor using dendritic cells simultaneously transfected with tumor RNA and granulocyte macrophage colony-stimulating factor RNA

• DOI: 10.1111/j.1349-7006.2007.00698.x
• 发表时间: 2008-02-01
• 期刊: CANCER SCIENCE
• 影响因子: 5.7
• 作者: Naka, Teiji;Iwahashi, Makoto;Yamaue, Hiroki
• 通讯作者: Yamaue, Hiroki

Streptococcal preparation OK-432 promotes the capacity of dendritic cells(DCs)to prime carcinoembryonic antigen(CEA)-specific cytotoxic T lymphocyte responses induced with genetically modified DCs that express CEA.

链球菌制剂 OK-432 可促进树突状细胞 (DC) 引发癌胚抗原 (CEA) 特异性细胞毒性 T 淋巴细胞反应的能力，这些反应由表达 CEA 的转基因 DC 诱导。

• 发表时间: 2008
• 期刊: Int J Oncol 32(2)
• 作者: Ojima T;et. al.;Koga K.et al.;Naka Teiji;Ojima Toshiyasu
• 通讯作者: Ojima Toshiyasu

Evaluation of chemosensitivity testing with highly purified tumor cells in 435 patients with gasytric cancer using an MTT assay.

使用 MTT 测定对 435 名胃癌患者进行高纯度肿瘤细胞的化疗敏感性测试进行评估。

• 发表时间: 2005
• 期刊: Anticancer Res 25(2A)
• 作者: Nakamura M;et. al.;Nakamura M et al.;Iwahashi M et al.;Noguchi K et al.
• 通讯作者: Noguchi K et al.

Successful cancer vaccine therapy for carcinoembryonic antigen(CEA)-expressing colon cancer using genetically modified dendritic cells that express CEA and Thelper-type1 cytokines in CEA transgenic mice

使用表达 CEA 和 Thelper-1 型细胞因子的转基因树突状细胞在 CEA 转基因小鼠中成功治疗表达癌胚抗原 (CEA) 的结肠癌

• 发表时间: 2007
• 期刊: Int J Cancer 120(3)
• 作者: Ojima T;et. al.;Koga K.et al.;Naka Teiji;Ojima Toshiyasu;Ojima Toshiyasu;Ojima Toshiyasu;Ojima Toshiyasu
• 通讯作者: Ojima Toshiyasu

OK-432 promotes the capacity of DCs to prime TAA-specific CTLs induced with genetically modified DCs expressing TAA

OK-432 促进 DC 启动表达 TAA 的转基因 DC 诱导的 TAA 特异性 CTL 的能力

• 发表时间: 2007
• 作者: Nakamura M;et. al.;Nakamura M et al.;Iwahashi M et al.;Noguchi K et al.;宮澤 基樹;宮澤 基樹;宮澤 基樹;Toshiyasu Ojima
• 通讯作者: Toshiyasu Ojima