Center for Biological Sequence Analysis, Department of Systems Biology, Technical University of Denmark, Søltofts plads, Building 224, 2800 Lyngby, Denmark.
J Immunol Methods. 2012 Dec 14;386(1-2):112-6. doi: 10.1016/j.jim.2012.09.005. Epub 2012 Sep 25.
Diverse methodologies ranging from experimental immunological studies to immunotherapy involve the application of human monocyte-derived dendritic cells (moDCs). Considerable donor-dependent variations in the moDC production of IL-12p70 affect the outcome of these methodologies. It has been shown that moDCs generated under standard conditions develop into two subsets based on CD1a-expression with the CD1a+ moDCs being the main IL-12p70 producers. This has however not been generally accepted, which we show here because the subset described as CD1a-negative does express CD1a, but at a lower level than the other subset. We further characterize the phenotype of these two subsets, showing that the CD1a-hi subset has a greater immunogenic phenotype, making this subset more suitable for immunotherapy. The two subsets have previously been separated by cell sorting, but as this technique is not available to many laboratories and has incompatibility with clinical settings, a more widely useable technique is warranted. Therefore we tested if magnetic-activated cell sorting is useful for the purpose, and show that it is possible to isolate IL-12p70-competent CD1a-hi moDCs to a <92% purity, irrespective of the starting purity.
从实验免疫学研究到免疫疗法，各种方法都涉及到人类单核细胞衍生的树突状细胞（moDC）的应用。IL-12p70 的 moDC 产生中相当大的供体依赖性变化会影响这些方法的结果。已经表明，在标准条件下产生的 moDC 根据 CD1a 表达分为两个亚群，其中 CD1a+ moDC 是主要的 IL-12p70 产生者。然而，这并没有被普遍接受，我们在这里展示的原因是，被描述为 CD1a-阴性的亚群确实表达 CD1a，但水平低于另一个亚群。我们进一步描述了这两个亚群的表型，表明 CD1a-hi 亚群具有更强的免疫原性表型，因此更适合免疫治疗。这两个亚群以前已经通过细胞分选进行了分离，但由于这种技术不是许多实验室都可用的，并且与临床环境不兼容，因此需要更广泛使用的技术。因此，我们测试了磁激活细胞分选是否适用于该目的，并表明无论起始纯度如何，都可以将具有 IL-12p70 能力的 CD1a-hi moDC 分离到 <92%的纯度。
