Alculumbre Solana, Pattarini Lucia
INSERM U932 Immunity and Cancer, 26 Rue D'Ulm, 75005, Paris, France.
Institut Curie, Centre de recherche, 26 Rue D'Ulm, 75005, Paris, France.
Methods Mol Biol. 2016;1423:153-67. doi: 10.1007/978-1-4939-3606-9_11.
Blood represents the most accessible source of human dendritic cells (DCs). We present here a method to isolate three DC subtypes, as identified until now, from peripheral blood: plasmacytoid dendritic cells (pDCs), CD141(+) myeloid DCs, and CD1c(+) myeloid DCs. The method is based on the sequential depletion of non-DCs. First, depletion of granulocytes, erythrocytes, and platelets is obtained by blood centrifugation over a Ficoll gradient. Then, antibodies recognizing non-DCs, combined with magnetic beads, allow enrichment of DCs from peripheral blood mononuclear cells (PBMCs). Finally, enriched DCs are purified and separated into the different subtypes by immunolabeling and fluorescence-activated cell sorting (FACS) using DC-specific surface markers.DC studies might contribute to the comprehension of human immune processes in physiological and pathological conditions. Human blood DCs targeting might be a useful tool to ameliorate inflammatory diseases and improve vaccination strategies.
血液是人类树突状细胞（DCs）最容易获取的来源。我们在此介绍一种从外周血中分离目前已确定的三种DC亚型的方法：浆细胞样树突状细胞（pDCs）、CD141(+)髓样DCs和CD1c(+)髓样DCs。该方法基于对非DCs的顺序去除。首先，通过在Ficoll梯度上进行血液离心来去除粒细胞、红细胞和血小板。然后，与磁珠结合的识别非DCs的抗体可从外周血单核细胞（PBMCs）中富集DCs。最后，通过使用DC特异性表面标志物进行免疫标记和荧光激活细胞分选（FACS），对富集的DCs进行纯化并分离为不同的亚型。DC研究可能有助于理解生理和病理条件下的人类免疫过程。靶向人类血液DCs可能是改善炎症性疾病和改进疫苗接种策略的有用工具。
