Ueda Yuji, Itoh Tsuyoshi, Fuji Nobuaki, Harada Sachio, Fujiki Hiroshi, Shimizu Keiji, Shiozaki Atsushi, Iwamoto Arihiro, Shimizu Takeshi, Mazda Osam, Kimura Takafumi, Sonoda Yoshiaki, Taniwaki Masafumi, Yamagishi Hisakazu
Department of Surgery, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, 465 Kajii-cho, Kawaramachi-Hirokoji, Kamigyo-ku, Kyoto 602-8566, Japan.
Cancer Immunol Immunother. 2007 Mar;56(3):381-9. doi: 10.1007/s00262-006-0197-8. Epub 2006 Jul 8.
Recent studies have suggested that dendritic cell (DC)-based immunotherapy is one promising approach for the treatment of cancer. We previously studied the clinical toxicity, feasibility, and efficacy of cancer vaccine therapy with peptide-pulsed DCs. In that study, we used granulocyte colony-stimulating factor (G-CSF)-mobilized peripheral blood monocytes as a cell source of DCs. However, previous investigations have suggested that G-CSF-mobilized peripheral blood monocytes produce reduced levels of proinflammatory cytokines such as interleukin (IL)-12 and tumor necrosis factor (TNF)-alpha. These T helper (Th)-1-type cytokines are thought to promote antitumor immune response. In this study, we assessed the functional abilities of DCs generated from G-CSF-mobilized monocytes obtained from 13 patients with CEA-positive advanced solid cancers. Peripheral blood mononuclear cells were obtained from leukapheresis products collected before and after systemic administration of G-CSF (subcutaneous administration of high-dose [5-10 microg/kg] human recombinant G-CSF for five consecutive days). In vitro cytokine production profiles after stimulation with lipopolysaccharide (LPS) were compared between monocytes with and without G-CSF mobilization. DCs generated from monocytes were also examined with respect to cytokine production and the capacity to induce peptide-specific T cell responses. Administration of G-CSF was found to efficiently mobilize peripheral blood monocytes. Although G-CSF-mobilized monocytes (G/Mo) less effectively produced Th-1-type cytokines than control monocytes (C/Mo), DCs generated from G/Mo restored the same level of IL-12 production as that seen in DCs generated from C/Mo. T cell induction assay using recall antigen peptide and phenotypic analyses also demonstrated that DCs generated from G/Mo retained characteristics identical to those generated from C/Mo. Our results suggest that G-CSF mobilization can be used to collect monocytes as a cell source for the generation of DCs for cancer immunotherapy. DCs generated in this fashion were pulsed with HLA-A24-restricted CEA epitope peptide and administered to patients safely; immunological responses were induced in some patients.
最近的研究表明,基于树突状细胞(DC)的免疫疗法是一种很有前景的癌症治疗方法。我们之前研究了肽脉冲DCs癌症疫苗疗法的临床毒性、可行性和疗效。在该研究中,我们使用粒细胞集落刺激因子(G-CSF)动员的外周血单核细胞作为DCs的细胞来源。然而,之前的研究表明,G-CSF动员的外周血单核细胞产生的促炎细胞因子水平降低,如白细胞介素(IL)-12和肿瘤坏死因子(TNF)-α。这些辅助性T细胞(Th)1型细胞因子被认为可促进抗肿瘤免疫反应。在本研究中,我们评估了从13例癌胚抗原(CEA)阳性晚期实体癌患者获取的G-CSF动员单核细胞产生的DCs的功能能力。外周血单个核细胞取自全身给予G-CSF(皮下连续5天给予高剂量[5-10μg/kg]人重组G-CSF)前后采集的白细胞分离产物。比较了有和没有G-CSF动员的单核细胞经脂多糖(LPS)刺激后的体外细胞因子产生谱。还检测了单核细胞产生的DCs的细胞因子产生情况以及诱导肽特异性T细胞反应的能力。发现给予G-CSF可有效动员外周血单核细胞。虽然G-CSF动员的单核细胞(G/Mo)产生Th-1型细胞因子的效率低于对照单核细胞(C/Mo),但G/Mo产生的DCs恢复了与C/Mo产生的DCs相同水平的IL-12产生。使用回忆抗原肽的T细胞诱导试验和表型分析也表明,G/Mo产生的DCs保留了与C/Mo产生的DCs相同的特征。我们的结果表明,G-CSF动员可用于收集单核细胞作为癌症免疫治疗中DCs产生的细胞来源。以这种方式产生的DCs用HLA-A24限制性CEA表位肽脉冲,并安全地给予患者;在一些患者中诱导了免疫反应。