源自脐带血和动员的成人外周血塑料贴壁单核细胞的体外扩增树突状细胞的比较：脐带血树突状细胞的同种异体反应性降低。

A comparison of ex vivo expanded DCs derived from cord blood and mobilized adult peripheral blood plastic-adherent mononuclear cells: decreased alloreactivity of cord blood DCs.

作者信息

Bracho F, van de Ven C, Areman E, Hughes R M, Davenport V, Bradley M B, Cai J-W, Cairo M S

机构信息

Department of Pediatrics and Lombardi Cancer Center, Georgetown University Hospital, Washington, DC, USA.

出版信息

Cytotherapy. 2003;5(5):349-61. doi: 10.1080/14653240310003017.

DOI:10.1080/14653240310003017

PMID:14578097

Abstract

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.

摘要

背景

脐血（CB）已被用作多种恶性和非恶性疾病可移植异基因干细胞的替代来源。然而，我们已经证明，无关脐血移植（UCBT）后T细胞和B细胞功能以及T细胞亚群的恢复延迟，并且脐血单个核细胞（MNC）在产生包括G-CSF、GM-CSF、M-CSF、IL-12和IL-15在内的细胞因子方面存在缺陷。在本研究中，我们研究了从脐血与动员的成人外周血（APB）体外生成树突状细胞（DC），以供日后用作过继性细胞免疫治疗。

方法

将脐血和APB贴壁单个核细胞在含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7天。对其形态、表型、免疫组织化学、集落形成活性和混合淋巴细胞反应（MLR）中的同种异体反应性进行评估。

结果

脐血和APB单核细胞来源的体外扩增DC表达相似的DC标志物CD83（31.27% + 11.7%对34.0% + 5.2%，脐血对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CD1a但不表达CD14。FLT-3配体和TGF-β均未增强DC的扩增。向脐血培养物中添加1OK的自体血浆可促进更高的细胞存活率，并使CD1a + /CD80 + 细胞回收率提高150%。在同种异体MLR中，脐血DC作为成人同种异体T细胞刺激物的效力是APB DC的62%（p < 0.05）。