BACKGROUND Cord blood (CB) has been used as an alternative source of transplantable allogeneic stem cells for a variety of malignant and non-malignant diseases. However, we have demonstrated delayed recovery of T- and B-cell function, and T-cell subsets post unrelated CB transplantation (UCBT), and deficiencies of CB mononuclear cells (MNC) in producing cytokines, including G-CSF, GM-CSF, M-CSF, IL-12, and IL-15. In this study we have investigated the ex vivo generation of DC from CB versus mobilized adult peripheral blood (APB) for later use as adoptive cellular immunotherapy. METHODS CB and APB-adherent MNC were cultured in serum-free media with GM-CSF IL-4, FLT-3 ligand, tumor growth factor-beta (TGF-beta), and tumor necrosis factor-alpha (TNF-alpha) for 7 days. Morphology, phenotype, immunohistochemistry, clonogenic activity, and alloreactivity in MLR were evaluated. RESULTS CB and APB monocyte-derived ex vivo expanded DC expressed similar DC markers CD83 (31.27+ 11.7% versus 34.0+ 5.2%, CB versus APB), CD1a (23.4+ 4.2% versus 27.6+ 6.3%), and CD80 (21.97+ 12.01% versus 27.7+ 5.95). Immunohistochemistry showed that cells with DC morphology expressed CDla but not CD14. Neither FLT-3 ligand nor TGF-fl enhanced DC expansion. Addition of 10% autologous plasma to CB cultures promoted greater cell survival and a 150% increase in CDla + /CD80+ cell recovery. CB DC were 62% as effective stimulators of adult allogeneic T-cels as APB DC (p < .05) in allogeneic MLR. DISCUSSION While phenotypically similar, CB and APB DC have differential potency in allogeneic MLR, which may account for the difference in GvHD and infection incidence and severity between UCBT and allogeneic stem cell transplantation, and may require a different approach for adoptive cellular immunotherapy. The mechanism(s) associated with these differences require further elucidation.

中文翻译：

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来自脐带血的体外扩增 DCs 和动员的成人外周血塑料粘附单核细胞的比较：脐带血 DCs 的同种异体反应性降低

背景脐带血(CB)已被用作用于多种恶性和非恶性疾病的可移植同种异体干细胞的替代来源。然而，我们已经证明 T 细胞和 B 细胞功能的恢复延迟，以及无关 CB 移植 (UCBT) 后的 T 细胞亚群，以及 CB 单核细胞 (MNC) 在产生细胞因子方面的缺陷，包括 G-CSF、GM-CSF 、M-CSF、IL-12和IL-15。在这项研究中，我们研究了从 CB 与动员的成人外周血 (APB) 离体生成 DC，以便以后用作过继细胞免疫疗法。方法 将 CB 和 APB 粘附的 MNC 在含有 GM-CSF IL-4、FLT-3 配体、肿瘤生长因子-β（TGF-β）和肿瘤坏死因子-α（TNF-α）的无血清培养基中培养，用于7天。形态学、表型、免疫组织化学、克隆活性、并评估了 MLR 中的同种异体反应性。结果 CB 和 APB 单核细胞衍生的体外扩增 DC 表达相似的 DC 标志物 CD83（31.27+ 11.7% 对 34.0+ 5.2%，CB 对 APB）、CD1a（23.4+ 4.2% 对 27.6+ 6.3%）和 CD80（21.97%） 12.01% 对 27.7+ 5.95）。免疫组织化学显示具有 DC 形态的细胞表达 CDla 但不表达 CD14。FLT-3 配体和 TGF-f1 均不增强 DC 扩张。向 CB 培养物中加入 10% 自体血浆促进了更高的细胞存活率和 150% 的 CDla + /CD80+ 细胞回收率增加。在同种异体 MLR 中，CB DC 是成人同种异体 T 细胞的 62% 有效刺激剂作为 APB DC (p < .05)。讨论 虽然表型相似，但 CB 和 APB DC 在同种异体 MLR 中具有不同的效力，这可能解释了 UCBT 和同种异体干细胞移植之间 GvHD 和感染发生率和严重程度的差异，并且可能需要不同的方法进行过继细胞免疫治疗。与这些差异相关的机制需要进一步阐明。