IMMUNE THERAPY FOR MALIGNANT GLIOMA USING GENETICALLY-MODIFIED TUMOR CELLS AND T-LYMPHOCYTES

使用基因修饰肿瘤细胞和 T 淋巴细胞对恶性神经胶质瘤进行免疫治疗

• 批准号: 07457306
• 负责人: HIRAKAWA Kimiyoshi
• 金额: $ 4.22万
• 依托单位: TOKYO MEDICAL AND DENTAL UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07457306/
• 关键词: MALIGNANT GLIOMA; TUMOR VACCINE; CYTOKINE; RETROVIRUS VECTOR; サイトカイン; レトロウイルス・ベクター

To realize a novel vaccination treatment for malignant gliomas using tumor cells genetically modified to express certain cytokines, it is essential to achieve an efficient gene transduction into primary cultured cells. We investigated the feasibility of preparing a glioma vaccine with retrovirus-mediated gene transduction. Glioma cells were cultured primarily from surgically resected tumor tissues of six patients. We obtained more than 1000-fold proliferation of cultures in all six cases. In vitro infection with a recombinant retrovirus GKlacZ marker gene resulted in over 65& gene transfer to the primary cultured glioma cells. Further enrichment of transduced cells was possible by employing repeated infections or using vector with neo^r marker gene. Two cytokine genes, GM-CSF and IL-4, were introduced into glioma cells by sequential transduction with two single-expression GK vectors. The transduced glioma cells produced high levels of both cytokines. Our data indicate the feasibility of retrovirus-mediated preparation of gene-modified tumor vaccines from clinical glioma materials, which could be useful potentiating antitumor immunity in glioma patients.

为了实现使用经基因修饰以表达某些细胞因子的肿瘤细胞的恶性胶质瘤的新型疫苗治疗，必须实现有效的基因转导到原代培养细胞中。我们探讨了逆转录病毒介导的基因转导制备脑胶质瘤疫苗的可行性。神经胶质瘤细胞培养主要从手术切除的肿瘤组织的6名患者。我们在所有六种情况下都获得了超过1000倍的培养增殖。用重组逆转录病毒GKlacZ标记基因体外感染原代培养的胶质瘤细胞，导致超过65个基因转移。通过重复感染或使用带有neo^r标记基因的载体，可以进一步富集转导的细胞。将两种细胞因子基因GM-CSF和IL-4通过两种单表达GK载体的顺序转导导入胶质瘤细胞。转导的胶质瘤细胞产生高水平的两种细胞因子。我们的数据表明，逆转录病毒介导的基因修饰的肿瘤疫苗的制备从临床胶质瘤材料的可行性，这可能是有用的增强抗肿瘤免疫在胶质瘤患者。

项目成果

Hiroaki Wakimoto, Yoko Yoshida, Masaru Aoyagi, Kimiyoshi Hirakawa, Hirofumi Hamada: "Efficient Retrovirus-mediated Cytokine-gene Transduction of Primary-cultured Human Glioma Cells for Tumor Vaccination Therapy" Japanese Journal of Cancer Research. (in pr

Hiroaki Wakimoto、Yoko Yoshida、Masaru Aoyagi、Kimiyoshi Hirakawa、Hirofumi Hamada：“原代培养的人胶质瘤细胞的高效逆转录病毒介导的细胞因子基因转导用于肿瘤疫苗接种治疗”日本癌症研究杂志。

Hiroaki Wakimoto et.al.: "Intensified Autitumor Immunity by a Cancer Vacccine That Produces Granlocyte-Macrophage Colony-stimulating Factor plus Interleukin 4" Cancer Research. 56. 1828-1833 (1996)

Hiroaki Wakimoto 等人：“通过产生粒细胞巨噬细胞集落刺激因子加白细胞介素 4 的癌症疫苗增强自身肿瘤免疫”癌症研究。

Hiroaki Wakimoto et.al.: "Efficient Retrovirus-mediated Cytokine-gene Transduction of Primary-cultured Human Glioma Cells for Tumor Vaccination Therapy" Japanese Journal of Cancer Research. (1997)

Hiroaki Wakimoto 等人：“原代培养的人胶质瘤细胞的高效逆转录病毒介导的细胞因子基因转导用于肿瘤疫苗接种治疗”日本癌症研究杂志。

Hiroaki Wakimoto,Junko Abe,Rikiya Tsunoda,Masaru Aoyagi,Kimiyoshi Hirakawa,Hirofumi Hamada: "Intensified Autitumor Immunity by Cancer Vaccine Producing GM・CSF plus IL-4" Cancer Research. (in press). (1996)

Hiroaki Wakimoto、Junko Abe、Rikiya Tsunoda、Masaru Aoyagi、Kimiyoshi Hirakawa、Hirofumi Hamada：“通过产生 GM·CSF 和 IL-4 的癌症疫苗增强自身肿瘤免疫”（出版中）。

Hiroaki Wakimoto: "Intensified Antitumo Immunity by Cancer Vaccine Producing GM-CSF plus IL-4" Cancer Research. (in press). (1996)

Hiroaki Wakimoto：“通过产生 GM-CSF 和 IL-4 的癌症疫苗增强抗肿瘤免疫”癌症研究。