Tumor specific immunotherapy using dendritic cells generated from allogeneic peripheral stem cell.

为了开发一种微创治疗晚期泌尿系统癌的策略，我们。研究了利用由亲属外周血单核细胞产生的树突状细胞（dc）进行细胞免疫治疗的潜力。2002年，我们合成了用于前列腺癌的HLA-A2和-A24限制性表位肽和用于膀胱癌的黑色素瘤抗原-3表位肽。负载各主要组织相容性抗原1类限制性表位肽的树突状细胞能够诱导相应的抗原特异性细胞毒性t淋巴细胞，能够在体外裂解靶癌细胞。使用装载这些表位肽的自体单核细胞来源的dc进行免疫治疗，在前一年至少引入HLA-A2或-A24阳性膀胱癌和前列腺癌患者，并在2003年第二财政年度进行。有患者显示转移性肿瘤的大小减少了约50%或ser…更多的肿瘤学标志物。所有患者均未观察到严重的不良反应。这些结果表明，基于DC的免疫治疗对常规治疗有耐药性的癌症患者是安全有效的。我们还利用重组cDNA表达克隆的血清学鉴定方法鉴定出新的膀胱癌特异性抗原，并合成了这些抗原的HLA -A24限制性表位肽。负载这些表位肽的dc可以在体外诱导HLA-A24阳性膀胱癌细胞裂解。本研究的主要目的是利用异体外周干细胞诱导dc，我们证明了用磁性细胞选择系统从外周血中分离出CD34阳性祖细胞，纯度为85.3%。这表明，该系统的应用使细胞调节更有效。此外，为了检验同种异体CD34祖细胞生成的dc对供体淋巴细胞的影响，我们采用混合淋巴细胞培养的方法，将双胞胎中一人的dc、另一人的淋巴细胞、父母和非近亲的dc进行混合培养实验。非近亲淋巴细胞对dc的排斥试验显示，刺激指数为3.6，与近亲组相当。这一结果表明，可能没有必要将dc的来源限制为来自近亲的CD34祖细胞。少

In order to develop a less invasive treatment strategy for advanced urological cancer, we. examined the potential of cell-immunotherapy using dendritic cells (DCs) generated from peripheral blood mono nuclear cells of kinsman. 2002 as a, we synthesized HLA-A2 and -A24 restricted epitope peptides prostate specific membrane antigen epitope peptide for prostate cancer and melanoma antigen-3 epitope peptide for bladder cancer. DCs loaded with each major histocompatibility antigen class 1 restricted epitope peptide were able to induce corresponding antigen- specific cytotoxic T-lymphocytes capable to lyse the target cancer cells in vitro. The immunotherapy using autologous monocyte-derived DCs loaded with these epitope peptides, which was introduced to at least either HLA-A2 or -A24 positive patients with bladder cancer and prostate cancer in preceding year, was carried out through second fiscal year 2003. There were patients showed a >50% reduction in the size of metastatic tumor or in ser … More ological tumor marker. No severe adverse effect was observed in all patients treated with this therapy. These results suggested that the DC based immunotherapy was safe and, effective for patients with cancer resistant to conventional treatment. We also identified novel cancer specific antigens of bladder cancer by using serologic identification of recombinant cDNA expression cloning method, and synthesized the HLA -A24 restricted epitope peptides of these antigens. It was possible to elicit HLA-A24 positive bladder cancer cell lysis by DCs loaded with these epitope peptides in vitro. As for the main purpose of this study, the induction of DCs from allogeneic peripheral stem cells, we demonstrated that CD34 positive progenitor cells were separated from peripheral blood with the purity of 85.3% by using magnetic cell selection system. This made it appear that the application of this system enables to make cell conditioning more efficient. Furthermore, to examine the effect of DCs generated from allogeneic CD34 progenitor cells to the donor lymphocytes, we carried out mixed culture experiment using DCs from one of twin, lymphocytes from the other of twin, parents and non-kinsman in the same way of mixed lymphocyte culture. The rejection test of DCs by the lymphocytes of non-kinsman showed that the stimulation index was 3.6 comparable to the results in kin group. This result suggested that it might not be necessary to limit the source of DCs to CD34 progenitor cells from kinsman. Less