Gene therapy for gastrointestinal tumors using antigen presenting cells activated with gene transfer

使用通过基因转移激活的抗原呈递细胞进行胃肠道肿瘤的基因治疗

• 批准号: 13671367
• 负责人: TAGAWA Masatoshi
• 金额: $ 2.62万
• 依托单位: Chiba Cancer Center Research Institute
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13671367/
• 关键词: Gene therapy; CD40 ligand; CD40; dendritic cells; protective immunity; IL-12; CD86; Mig

We examined whether professional antigen presenting cells (dendritic cells) could be activated by the expression of CD40 ligand (CD40L) gene on tumors and antitumor immunity was subsequently induced. We retrovirally transduced tumors with the CD40L gene and established CD40L-expressed tumors whose MHC class I expression remained the same as that of parent tumors. Inoculation of the transduced cells into syngeneic mice revealed that growth of CD40L-expressed tumors was significantly retarded compared with that of parent tumors. Some of the mice completely rejected the CD40L-expressed tumors and the mice developed antigen-specific protective immunity. When nude mice were inoculated with the CD40L-expressed tumors, the growth was not different from that of parent tumors. In vitro culture of the CD40L-expressed tumors and bone marrow-derived dendritic cells showed that the cluster formation between dendritic cells and the CD40L-expressed but not parent tumors. The expression of MHC class II and activation marker CD86 on dendritic cells was upregulated after the coculture with CD40L-expressed but not parent tumors. The activated dendritic cells secreted IL-12, IL-18, IL-23 and Mig. These data collectively suggest that CD40L on tumors can activate dendritic cells through CD40/CD40L interaction and induce the expression of cytokines and chemokines, which play a crucial role in the generation of T cell-mediated antitumor effects.

我们检测了肿瘤表面CD40配体(CD40L)基因的表达能否激活专业抗原提呈细胞(树突状细胞)，从而诱导抗肿瘤免疫。我们逆转录病毒转导了CD40L基因的肿瘤，建立了CD40L表达的肿瘤，其MHC-I类表达与亲本肿瘤相同。将转导细胞接种到同基因小鼠体内，发现CD40L表达的肿瘤与亲本肿瘤相比生长明显减慢。部分小鼠完全排斥CD40L表达的肿瘤，并产生抗原特异性保护性免疫。将CD40L表达的肿瘤接种于裸鼠，其生长情况与亲本肿瘤无明显差异。体外培养的CD40L表达的肿瘤和骨髓来源的树突状细胞显示，树突状细胞和CD40L表达的肿瘤之间形成簇，但不表达亲本肿瘤。与CD40L-表达的肿瘤共培养后，树突状细胞表面MHC-II类分子和活化标记CD86的表达上调，但不表达亲本肿瘤。活化的树突状细胞分泌IL-12、IL-18、IL-23和Mig。提示肿瘤表面的CD40L可通过CD40/CD40L相互作用激活树突状细胞，诱导细胞因子和趋化因子的表达，在T细胞介导的抗肿瘤效应的产生中起关键作用。

项目成果

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Tomizawa M, tagawa M, et al.: "Decreased expression of the CCAAT/enhancer binding protein α gene involved in hepatocyte proliferation in human hepatocellular carcinoma"Int.J.Mol.Med.. 9. 597-600 (2002)

Tomizawa M、takawa M等人：“人肝细胞癌中参与肝细胞增殖的CCAAT/增强子结合蛋白α基因的表达降低”Int.J.Mol.Med..9.597-600(2002)

Tada, Y., O-Wang, J., Takenaga, K., Takiguchi, Y., Tatsumi, K., Kuriyama, T. and Tagawa, M.: "Expression of the TNF-α gene on mouse lung carcinoma cells suppresses spontaneous lung metastasis without affecting tumorigenicity"Oncol. Rep.. 9. 585-588 (2002)

Tada, Y.、O-Wang, J.、Takenaga, K.、Takiguchi, Y.、Tatsumi, K.、Kuriyama, T. 和 Takawa, M.：“TNF-α 基因在小鼠肺癌细胞上的表达抑制自发性肺转移而不影响致瘤性“Oncol. Rep.. 9. 585-588 (2002)