Efficient presentation of tumor idiotype to autologous T cells by CD83+ dendritic cells derived from highly purified circulating CD14+ monocytes in multiple myeloma patients
Efficient presentation of tumor idiotype to autologous T cells by CD83+ dendritic cells derived from highly purified circulating CD14+ monocytes in multiple myeloma patients
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DOI：
10.1016/s0301-472x(00)00486-0
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发表时间：
2000-08-01
影响因子：
2.6
通讯作者：
Lemoli, RM
中科院分区：
文献类型：
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作者：
Ratta, M;Curti, A;Lemoli, RM
目标.为了产生源自循环CD 14（+）细胞的成熟和全功能的CD 83（+）树突状细胞，所述循环CD 14（+）细胞高度纯化自多发性骨髓瘤患者的白细胞去除术产物，材料和方法。用高梯度磁分离法分离CD 14（+）单核细胞，用粒细胞-巨噬细胞集落刺激因子（GM-CSF）和白细胞介素-4（IL-4）诱导分化为未成熟的树突状细胞（DC）6-7天，再用肿瘤坏死因子-α（TNF-α）诱导至终末成熟。或用CD 40配体刺激，树突状细胞通过免疫表型、可溶性抗原摄取、细胞因子分泌、刺激同种异体T细胞的能力以及向自体T淋巴细胞呈递名义抗原（包括肿瘤独特型）的能力来表征。表型分析显示，在粒细胞-巨噬细胞集落刺激因子和白细胞介素-γ治疗后，90% +/- 6%的细胞恢复。4刺激表达未成熟树突状细胞典型的所有表面标志物，并证明摄取可溶性抗原的高能力，如FITC-葡聚糖测定所示。随后暴露于成熟刺激诱导CD 1a下调和CD 83，HLA-DR，共刺激分子并诱导大量白细胞介素-12的分泌成熟的CD 83（+）细胞显示抗原摄取能力降低，而它们在混合淋巴细胞反应中被证明是同种异体T细胞的有效刺激物，单核细胞来源的树突状细胞，在加入成熟刺激物之前脉冲，能够将可溶性蛋白如钥孔血蓝蛋白和破伤风类毒素呈递给自体T细胞，分别用于初次和二次免疫应答。相反，成熟的（CD 83（+））树突状细胞的脉冲诱导T细胞增殖的效率较低。更重要的是，CD 14（+）细胞来源的树突状细胞刺激自体T细胞增殖，以响应肿瘤抗原，如患者特异性独特型。此外，独特型脉冲的树突状细胞诱导纯化的CD 4（+）细胞分泌白细胞介素-2和γ-干扰素，当使用Fab免疫球蛋白片段时比使用完整蛋白时更好地实现T细胞活化。当分析来自健康志愿者的CD 14（+）细胞衍生的树突状细胞时，我们没有发现与来自骨髓瘤患者的样品在细胞产量、表型特征和功能特征方面有任何差异。这些研究表明，动员的纯化CD 14（+）细胞代表了生产适用于多发性骨髓瘤临床试验的成熟CD 83（+）树突状细胞同质细胞群的最佳来源，（C）2000国际实验血液学学会。出版社：Elsevier Science Inc.出版社：Elsevier Science Inc.
Objectives. To generate mature and fully functional CD83(+) dendritic cells derived from circulating CD14(+) cells highly purified from the leukapheresis products of multiple myeloma patients,Materials and Methods. CD14(+) monocytes mere selected by high-gradient magnetic separation and differentiated to immature dendritic cells with granulocyte-macrophage colony-stimulating factor and interleukin-4 For 6-7 days and then induced to terminal maturation by the addition of tumor necrosis factor-alpha. or stimulation with CD40 ligand, Dendritic cells were characterized by immunophenotyping, evaluation of soluble antigens uptake, cytokine secretion, capacity of stimulating allogeneic T cells, and ability of presenting nominal antigens, including tumor idiotype, to autologous T lymphocytes.Results, Phenotypic analysis showed that 90% +/- 6% of cells recovered after granulocyte-macrophage colony-stimulating factor and interleukin-4 stimulation expressed all surface markers typical of immature dendritic cells and demonstrated a high capacity of uptaking soluble antigens as shown by the FITC-dextran assay, Subsequent exposure to maturation stimuli induced the downregulation of CD1a and upregulation of CD83, HLA-DR, costimulatory molecules and induced the secretion of large amounts of interleukin-12 Mature CD83(+) cells showed a diminished ability of antigen uptake whereas they proved to be potent stimulators of allogeneic T cells in a mixed lymphocyte reaction, Monocyte-derived dendritic cells, pulsed before the addition of maturation stimuli, were capable of presenting soluble proteins such as keyhole limpet hemocyanin and tetanus toxoid to autologous T cells for primary and secondary Immune response, respectively. Conversely, pulsing of mature (CD83(+)) dendritic cells was less efficient for the induction of T-cell proliferation. More importantly, CD14(+) cells-derived dendritic cells stimulated autologous T-cell proliferation in response to a tumor antigen such as the patient-specific idiotype, Moreover, idiotype-pulsed dendritic cells induced the secretion of interleukin-2 and gamma-interferon by purified CD4(+) cells, T-cell activation was better achieved when Fab immunoglobulin fragments were used as compared with the whole protein. When dendritic cells derived from CD14(+) cells from healthy volunteers were analyzed, we did not find any difference with samples from myeloma patients as for cell yield, phenotypic profile, and functional characteristics.Conclusion. These studies demonstrate that mobilized purified CD14(+) cells represent the optimal source for the production of a homogeneous cell population of mature CD83(+) dendritic cells suitable for clinical trials in multiple myeloma, (C) 2000 International Society for Experimental Hematology. Published by Elsevier Science Inc. Published by Elsevier Science Inc.