Characterization of human blood dendritic cell subsets
Characterization of human blood dendritic cell subsets
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DOI：
10.1182/blood-2001-11-0097
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发表时间：
2002-12-15
期刊：
影响因子：
20.3
通讯作者：
Hart, DNJ
中科院分区：
文献类型：
--
作者：
MacDonald, KPA;Munster, DJ;Hart, DNJ
树突状细胞（DC）是刺激免疫应答的关键抗原呈递细胞，目前正在临床环境中进行研究。尽管定义为谱系阴性（Lin（-））HLA-DR+细胞，但这些制剂中的显著异质性是明显的，特别是在包含或排除CD 14（+）、CD 16（+）和CD 2（+）细胞方面。本研究使用流式细胞术和一组单克隆抗体（mAb）（包括第7届白细胞分化抗原研讨会的试剂）来确定2种标准化外周血单核细胞（PBMC）来源的Lin（-）HLA-DR+制剂的细胞组成。通过用CD 3、CD 14、CD 19、CD 11b和CD 16或CD 56 mAb去除PBMC制备Lin（-）细胞。对CD 16-充满的制备物的分析将Lin-HLA-DR+群体分成5个不重叠的亚群（平均值± 1 SD）：CD 123（平均值= 18.3% ± 9.7%）、CD 1b/c（18.6% ± 7.6%）、CD 16（49.6% ± 8.5%）、BDCA-3（2.7% ± 1.4%）和CD 34（5.0% ± 2.4%）。这5个亚群在相互比较、单核细胞和单核细胞衍生的DC（MoDC）时具有不同的表型。CD 85家族、C型凝集素、共刺激分子和分化/活化分子也在5个Lin-HLA-DR+亚群、单核细胞和MoDC上差异表达。CD 123（+）DC在体外的存活率较低，而CD 16（+）CD 11 c（+）DC在体外的存活率也较低。最后，在同种异体混合白细胞反应中用作刺激剂的个体亚群根据其同种异体刺激能力排序为CD 1b/c > CD 16> BDCA-3 > CD 123> CD 34。这些数据提供了一个机会，以标准化用于未来的分子，功能，甚至可能治疗研究的DC群体。
Dendritic cells (DCs) are key antigen-presenting cells for stimulating immune responses and they are now being investigated in clinical settings. Although defined as lineage-negative (Lin(-)) HLA-DR+ cells, significant heterogeneity in these preparations is apparent, particularly in regard to the inclusion or exclusion of CD14(+), CD16(+), and CD2(+) cells. This study used flow cytometry and a panel of monoclonal antibodies (mAbs), including reagents from the 7th Leukocyte Differentiation Antigen Workshop, to define the cellular composition of 2 standardized peripheral blood mononuclear cell (PBMCs)-derived Lin(-) HLA-DR+ preparations. Lin(-) cells were prepared from PBMCs by depletion with CD3, CD14, CD19, CD11b, and either CD16 or CD56 mAbs. Analysis of the CD16-replete preparations divided the Lin- HLA-DR+ population into 5 nonoverlapping subsets (mean +/- 1 SD): CD123 (mean = 18.3% +/-9.7%), CD1b/c (18.6% +/- 7.6%), CD16 (49.6% +/- 8.5%), BDCA-3 (2.7% +/- 1.4%), and CD34 (5.0% +/- 2.4%). The 5 subsets had distinct phenotypes when compared with each other, monocytes, and monocyte-derived DCs (MoDCs). The CD85 family, C-type lectins, costimulatory molecules, and differentiation/activation molecules were also expressed differentially on the 5 Lin- HLA-DR+ subsets, monocytes, and MoDCs. The poor viability of CD123(+) DCs in vitro was confirmed, but the CD16(+) CD11c(+) DC subset also survived poorly. Finally, the individual subsets used as stimulators in allogeneic mixed leukocyte reactions were ranked by their allostimulatory capacity as CD1b/c > CD16 > BDCA-3 > CD123 > CD34. These data provide an opportunity to standardize the DC populations used for future molecular, functional and possibly even therapeutic studies.