Comparison of human plasmacytoid dendritic cells and monocyte-derived dendritic cells. Comparison of human plasmacytoid dendritic cells and monocyte-derived dendritic cells

Transcriptome analysis was conducted to investigate the gene expression profiles of pDCs using bulk RNA-sequencing (RNA-seq). Peripheral blood mononuclear cells (PBMCs) from healthy donors (n=3) were stained with lineage markers (CD3, CD14, CD16, CD19, and CD56), and pDCs were identified via flow cytometry (fluorescence-activated cell sorting [FACS]) based on the co-expression of IL-3R (CD123) and BDCA-2 (CD303). mDCs were identified using CD11c and sorted from the same PBMC donor as a control. After sorting, mRNA was extracted from the sorted cells, including mDCs and pDCs. The whole transcriptome profile was analyzed via RNA-seq.

本研究通过批量RNA测序（RNA-seq）对浆细胞样树突状细胞（pDCs）的基因表达谱开展转录组分析。选取3名健康志愿者的外周血单个核细胞（PBMCs），采用谱系标志物（CD3、CD14、CD16、CD19及CD56）进行染色，随后通过流式细胞术（荧光激活细胞分选[FACS]），依据IL-3受体（CD123）与BDCA-2（CD303）的共表达特征鉴定pDCs。以CD11c为标志物鉴定髓系树突状细胞（mDCs），并从同一份供体的PBMC样本中分选得到作为对照。分选完成后，从包括mDCs与pDCs在内的分选细胞中提取信使RNA（mRNA），通过RNA-seq分析全转录组表达谱。