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

为了开发一种侵袭性较小的晚期泌尿系癌症的治疗策略，我们。研究了利用亲属外周血单个核细胞产生的树突状细胞(DC)进行细胞免疫治疗的可能性。2002年，我们合成了针对前列腺癌的人类白细胞抗原A2和A24限制性表位多肽和针对膀胱癌的黑色素瘤抗原3表位。负载各主要组织相容性抗原1类限制性表位的DC在体外均能诱导相应的抗原特异性细胞毒T淋巴细胞对靶细胞产生杀伤作用。自体单核细胞来源的树突状细胞负载这些表位多肽的免疫治疗，在前一年至少被引入到至少HLA-A2或-A24阳性的膀胱癌和前列腺癌患者中，一直持续到2003第二财年。有些患者的转移瘤大小或严重…缩小了50%。更多的肿瘤标志物。所有接受该疗法的患者均未见严重不良反应。这些结果表明，DC为基础的免疫治疗对常规治疗无效的癌症患者是安全有效的。我们还利用重组表达克隆的血清学鉴定方法鉴定了新的膀胱癌肿瘤特异性抗原，并合成了这些抗原的人类白细胞抗原-A24限制性表位多肽。负载这些表位多肽的DC在体外可诱导人类白细胞抗原A24阳性的膀胱癌细胞裂解。对于本研究的主要目的，异基因外周血树突状细胞的诱导，我们利用磁性细胞筛选系统从外周血中分离出CD34阳性祖细胞，纯度为85.3%。这使得这一系统的应用使细胞调理变得更有效。此外，为了检测同种异体CD34祖细胞产生的DC对供者淋巴细胞的影响，我们用双胞胎之一的DC、双胞胎之一的淋巴细胞、双亲和非亲属的淋巴细胞以相同的混合淋巴细胞培养方法进行了混合培养实验。非亲属组淋巴细胞对DC的排斥反应显示，刺激指数为3.6，与亲缘关系组相当。这一结果提示，可能没有必要将DC的来源限制为来自亲属的CD34祖细胞。较少

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