Establisment of tumor vaccine using autologous dendritic cells and its application for addaptive immunotherapy

自体树突状细胞肿瘤疫苗的研制及其在适应性免疫治疗中的应用

• 批准号: 09671321
• 负责人: MORISAKI Takashi
• 金额: $ 2.11万
• 依托单位: KYUSHU UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09671321/
• 关键词: dendritic cells; tumor vaccine; colon cancer; 癌特異的免疫療法

Dendritic cells (DC) possess the most potent antigen-prersenting activity among various antigen presenting cells. This study aims preclinical application of tumor vaccine using autologous dendritic cells. We tried to establish autologous dendritic cell vaccine using autologous colon cancer cells and dendritic cells, and focused on investigation of the methods for effective induction of autolpogpus tumor spesific T cells and culture system for tumor-spesific dendritic cells. We first established colon carcinoma cell line, CE-1, from the 38 years. male patient with advanced colon cancer. These tumor cells revealed HLA-type Class I A31, and A11. We tried to establish tumor-spesific adendritic cells using autologouis cells and HLA-matched healthy volunteer. 1-2 x 10^6 dendritic cells were obtained from 100 ml of heparinized peripheral blood and culture media containing IL-4 and GM-CSF.When irradiated or freezing-thawing tumor cells were added to the dendritic cells and furtherly cultured in the presence of IL-4 and GM-CS F, tumor-spesific dendritic cells were obtained. These cells were positive for expression of IL-12 and IFN-gamma mRNA and the expression of HLA-DR, CD80, and ICAM-1.When autologous T cells were added to the tumor-pulsed dendritic cells and cultured in the presence of IL-2 and IL-4 for subsequent 2 weeks, autologous tumor-reactive CTL were obtained. Because theses T cell possess cytotoxic activity against autologous tumor cells releasing IFN-gamma. However we failed in culture of dendritic cells in a large number using gellatin-beads, which was one of aims of this study. Further studies are needed to establish and verify effective systems for culture of autologpus-tumor pulsed dendritic cell vaccine.

树突状细胞（DC）是目前发现的具有最强抗原提呈活性的细胞。本研究旨在利用自体树突状细胞进行肿瘤疫苗的临床前应用。本研究尝试利用自体结肠癌细胞和树突状细胞构建自体树突状细胞疫苗，并着重探讨自体肿瘤特异性T细胞的有效诱导方法和肿瘤特异性树突状细胞的培养体系。38年来，我们首次建立了结肠癌细胞系CE-1。晚期结肠癌的男性患者。这些肿瘤细胞显示HLA型I类A31和A11。我们尝试用自体细胞和HLA匹配的健康志愿者建立肿瘤特异性的腺样细胞。1-2从100 ml肝素化的外周血和含IL-4和GM-CSF的培养液中获得10 × 10^6个树突状细胞，将照射或冻融的肿瘤细胞加入树突状细胞中，在IL-4和GM-CSF存在下进一步培养，获得肿瘤特异性树突状细胞。这些细胞表达IL-12和IFN-γ mRNA，表达HLA-DR、CD 80和ICAM-1。将自体T细胞加入肿瘤致敏的树突状细胞中，在IL-2和IL-4存在下培养2周，可获得自体肿瘤反应性CTL。因为这些T细胞对释放IFN-γ的自体肿瘤细胞具有细胞毒性活性。本研究的目的之一是利用明胶微球进行树突状细胞的大量培养，但未能成功。建立和验证有效的自体肿瘤致敏树突状细胞疫苗培养体系尚需进一步研究。