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Establishment of dendritic cell immunotherapy based on the chemokine expression profile of cancer cells

基于癌细胞趋化因子表达谱建立树突状细胞免疫疗法

基本信息

批准号：
14370357
负责人：
TSUJITANI Shunichi
金额：
$ 7.87万
依托单位：
National University Corporation Tottori University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2004
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14370357/
关键词：
Cancer Dendritic cell Electrofusion Tumor antigen Vaccine ケモカイン遊走能消化器癌

项目摘要

Some previous reports indicated that fusion cells of DCs and tumor cells could function as antigen-presenting cells(APCs) containing tumor specific antigens. Several clinical trials with DC/Tumor fusion cells demonstrated an obvious tumor regression in patients with advanced stage of disease. In our study, the DCs and irradiated tumor cells were mixed at a ratio of 1:1 and incubated in serum-free RPMI 1640 medium containing 50% polyethylene glycol(PEG) for 1 min to induce cell fusion. Then, we used electrofusion techniques for producing the fusion cells. In a preliminary study, the optimal cell numbers (1.0x10^6 of DCs : 1.0x 10^6 of tumor cells/mL) and alignment voltage (100V for 10sec) were determined. Cell fusion was performed using ECM2001 electrofusion apparatus and confirmed by fluorescent microscopy and flow cytometry. The DCs were successfully fused with the allogeneic gastric cancer cells resulting in hybrid cells that functioned as antigen-presenting cells, because they induced allogeneic CD4+ T cell proliferation. A gastric cancer cell line, MKN-45, and DC hybrid cell induced autologous CD4+ and CD8+ T cell proliferation, indicating that MKN-45-DC hybrids present some antigens from the MKN-45 cells to the CD4+ and CD8+ T cells. CD8+ T cells stimulated by the MKN-45-DC hybrid cells were able to kill MKN-45 when used for immunization. The CTLs killed another gastric cancer cell line, MKN-1, as well as a melanoma cell line, 888 mel, suggesting the recognition of a shared tumor antigen. MKN-45 specific CTLs can recognize carcinoembryonic antigen(CEA), indicating that the killing is due to tumor antigens as well as alloantigens. Further studies are requested for practical use of DC/tumor fusion cells in cancer therapy. Moreover, the mechanisms of counterattack by tumor cells against immune cells should be clarified to establish an effective DC treatment for cancer.

之前的一些报道表明，DC和肿瘤细胞的融合细胞可以充当含有肿瘤特异性抗原的抗原呈递细胞（APC）。 DC/肿瘤融合细胞的多项临床试验表明，晚期疾病患者的肿瘤明显消退。在我们的研究中，DC和照射后的肿瘤细胞以1:1的比例混合，并在含有50%聚乙二醇（PEG）的无血清RPMI 1640培养基中孵育1分钟以诱导细胞融合。然后，我们使用电融合技术来生产融合细胞。在初步研究中，确定了最佳细胞数量（1.0x10^6 的 DC：1.0x 10^6 肿瘤细胞/mL）和对准电压（100V，10秒）。使用ECM2001电融合仪进行细胞融合，并通过荧光显微镜和流式细胞术确认。 DCs 与同种异体胃癌细胞成功融合，产生了充当抗原呈递细胞的杂交细胞，因为它们诱导同种异体 CD4+ T 细胞增殖。胃癌细胞系 MKN-45 和 DC 杂交细胞诱导自体 CD4+ 和 CD8+ T 细胞增殖，表明 MKN-45-DC 杂交细胞将 MKN-45 细胞的一些抗原呈递给 CD4+ 和 CD8+ T 细胞。 MKN-45-DC 杂交细胞刺激的 CD8+ T 细胞在用于免疫时能够杀死 MKN-45。 CTL 杀死了另一种胃癌细胞系 MKN-1 以及黑色素瘤细胞系 888 mel，这表明它们识别了共同的肿瘤抗原。 MKN-45特异性CTL可以识别癌胚抗原(CEA)，表明杀伤作用是由于肿瘤抗原以及同种异体抗原。 DC/肿瘤融合细胞在癌症治疗中的实际应用需要进一步的研究。此外，应阐明肿瘤细胞对免疫细胞的反击机制，以建立有效的 DC 癌症治疗方法。