Gene therapy for cancer using intratumoral injection of dendritic cells genetically modified to express interleukin 12

使用经过基因改造表达白细胞介素 12 的树突状细胞瘤内注射进行癌症基因治疗

• 批准号: 12671145
• 负责人: ICHIKAWA Naoya
• 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: 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-12671145/
• 关键词: gene therapy; viral vector; dendritic cell; OK-432; interleukin-12; インターロイキン12

Dendritic cells (DCs) are potent professional antigen presenting cells and play a key role in cellular immunity. We have previously demonstrated that specific antitumor immune response can be induced with intratumoral (I.t.) injection of bone marrow-derived DCs genetically engineered with interleukin (IL)-12 genes. In this study, we examined whether DCs stimulated with OK-432 could be used in the same manner. In vivo administration of recombinant IL-12 causes NK cells and T cells to secrete IFN-g and enhances the cytolytic functions against tumor cells. These cytokines, however, induce abundant secretion of nitric oxide (NO) from activated macrophages, and appear to suppress cellular immunity in murine system. OK-432 has been clinically used as a potent biological modifiers for treating cancer patients and is known to induce multiple cytokines including IFN-g and IL-12 in vitro. In intradermal tumor modeles using MCA205 into C57BL/6 mice, I.t. injection bone marrow-derived DCs with or without OK-432 showed marginal antitumor effects on day 7 established tumors. However, potent antitumor effects were observed when they are treated with the combination of DCs, OK-432, and N-nitro-L-arginine methyl ester (L-NAME), which inhibits inducible nitric oxide synthase (iNOS)-mediated NO production. Furthermore, tumor specific and potent cytotoxic T lymphocytes (CTLs) could be obtained from the splenocytes of the mice treated with this combination. These results suggested that tumor specific cellular immunity in mice could be strongly suppressed with NO, which is abundantly secreted by macrophages and DCs stimulated with OK-432. Thus, I.t. injection of DCs and OK-432 could be a promisingly cancer immunotherapy if NO production is properly regulated.

树突状细胞（Dendritic cells，DC）是一种功能强大的专职抗原提呈细胞，在细胞免疫中起着重要作用。我们先前已经证明，特异性抗肿瘤免疫应答可以用肿瘤内（I.t.）注射用白细胞介素（IL）-12基因遗传工程化的骨髓来源的DC。在这项研究中，我们研究了是否可以以相同的方式使用OK-432刺激的DC。重组IL-12的体内给药引起NK细胞和T细胞分泌IFN-γ并增强对肿瘤细胞的细胞溶解功能。然而，这些细胞因子诱导激活的巨噬细胞分泌大量的一氧化氮（NO），并似乎抑制小鼠系统的细胞免疫。OK-432已在临床上用作治疗癌症患者的有效生物调节剂，已知在体外诱导多种细胞因子，包括IFN-g和IL-12。在使用MCA 205进入C57 BL/6小鼠的皮内肿瘤模型中，I.t.注射有或没有OK-432的骨髓来源的DC对第7天建立的肿瘤显示出边缘抗肿瘤作用。然而，当用DC、OK-432和N-硝基-L-精氨酸甲酯（L-NAME）的组合处理时，观察到有效的抗肿瘤作用，所述组合抑制诱导型一氧化氮合酶（iNOS）介导的NO产生。此外，可以从用该组合处理的小鼠的脾细胞获得肿瘤特异性和有效的细胞毒性T淋巴细胞（CTL）。这些结果表明，小鼠中的肿瘤特异性细胞免疫可以被NO强烈抑制，所述NO由用OK-432刺激的巨噬细胞和DC大量分泌。因此，I.t.如果适当调节NO的产生，DC和OK-432的注射可能是有希望的癌症免疫疗法。

项目成果

市川直哉, 田原秀晃: "樹状細胞とサイトカインを用いた癌免疫療法"血液・腫瘍科. 43・6. 453-457 (2001)

Naoya Ichikawa，Hideaki Tahara：“使用树突状细胞和细胞因子的癌症免疫治疗”血液学和肿瘤学43・6（2001）。

Naoya Ichikawa: "Cancer immunotherapy using dendritic cells with cytokines"Hematology and oncology. 43(6). 453-457 (2001)

Naoya Ichikawa：“使用树突状细胞和细胞因子进行癌症免疫治疗”血液学和肿瘤学。