Clinical trial of specific immuno-therapy against patients with advanced solid cancer including lung cancer using mature dendritic cells

树突状细胞(DC)是最强大的抗原提呈细胞。肿瘤相关抗原肽(TAA)致敏的树突状细胞已用于肿瘤免疫治疗。早期的临床研究证明了这些试验的安全性，但临床效果还不够。大多数研究使用外周血单核细胞生成的未成熟DC与粒细胞-巨噬细胞集落刺激因子(GM-CSF)和白介素4(IL-4)联合使用。在这里，我们用链球菌衍生物OK-432诱导的成熟DC进行了主动免疫治疗的I期临床试验。外周血单核细胞与GM-CSF、IL-4共同培养6天，再与GM-CSF、IL-4、OK-432共同培养2天，获得DC。在注射前，DC以HLA-A^*2402限制性MAGE-3多肽(IMPKAGLLI)冲击DC，以锁孔帽血蓝蛋白(KLH)为对照抗原。我们选择了有MAGE-3mRNA表达的晚期实体瘤患者的HLA-A^*2402阳性患者。DC疫苗皮下注射…每2周1次，共接种4次，剂量递增设计，每队列剂量水平分别为0.1(组1)、0.3(组2)和1(组3)×10^8 DC/注。每组3名患者。采用迟发型超敏反应(DTH)和MHC四聚体进行免疫学监测。这一协议得到了很好的容忍。所有患者均有轻度发热(1~2级)和注射部位局部反应(1级红斑和硬结)。MAGE-3多肽的DTH在高剂量DC作用的早期呈阳性反应。1×10~(-8)DC/次注射能最有效地诱导DTH阳性反应。肿瘤标志物(CEA)下降1例。1名患者病情稳定(SD)达4个月，但其他患者病情进展。这些结果表明，成熟DC接种是安全可行的，但要获得客观的临床反应还需要进一步的研究。为了提高DC疫苗的临床效果，寻找在肺癌中表达的新的肿瘤抗原，更有效地诱导肿瘤特异性免疫是途径之一。我们找到了这些肿瘤抗原的候选抗原HM1.24。此外，我们还鉴定了HM1.24抗原的肿瘤多肽，它可与人类白细胞抗原A24结合，并能诱导细胞毒性T淋巴细胞。使用HM 1.24多肽进行临床试验是意料之中的。较少

Dendritic cells (DCs) are the most potent antigen-presenting cells. DCs pulsed with peptides of tumor-associated antigens (TAA) have been used in cancer immunotherapy. An early clinical study demonstrated the safety of these trials, but the clinical effect was not sufficient. Most studies have used immature DCs generated from peripheral blood monocytes with granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-4 (IL-4). Here, we conducted phase I clinical trial of active immunotherapy using mature DCs induced by a streptococcus derivatives OK-432. DCs were generated from blood monocytes by culturing with GM-CSF and IL-4 for 6 days and then GM-CSF,IL-4 and OK-432 for 2 days. Before injection, DCs were pulsed with MAGE-3 peptide (IMPKAGLLI), which is restricted for HLA-A^*2402, and keyhole limpet hemocyanin (KLH) as a control antigen. We selected HLA-A^*2402-positive patients who had advanced solid tumors expressing MAGE-3 mRNA. DC vaccine was administered subcutaneou … More sly every 2 weeks for a total of four vaccinations in a dose-escalation design at the dose level per cohort of 0.1 (Group 1),0.3 (Group 2) and 1 (Group 3) x 10^8 DCs/injection. Three patients were enrolled in each group. Immunological monitoring with delayed type hypersensitivity (DTH) reaction and MHC tetramer was performed. This protocol was well tolerated. A mild fever (Grade 1 to 2) and local reaction of injection site (erythema and induration : Grade 1) were found in all patients. DTH for MAGE-3 peptide became to be positive in the earlier period when a high dose of DCs was administered. 1 x 10^8 DCs/injection was most effective to induce a positive DTH reaction. The decrease of tumor marker (CEA) was found in one patient. One patient showed the stable disease (SD) for four months, but others were progressive. These results indicated that vaccination with mature DCs was safe and feasible, but further studies were required to obtain the objective clinical responses.To enhance the clinical effects of DC vaccine, one of the approaches was to find the novel tumor antigen expressing in lung cancer which more effectively induce tumor specific immunity. We found the candidate for these tumor antigens, HM1.24 antigen. Furthermore, we identified the tumor peptide of HM1.24 antigen which are restricted to HLA-A24 and can induce cytotoxic T lymphocytes. The clinical trial using HM 1.24 peptide was expected. Less