Identification and enumeration of dendritic cells in peripheral blood and bone marrow of healthy individuals / 中国实验血液学杂志

ABSTRACT

The study was aimed to analyse and enumerate the dendritic cells (DC) subsets in peripheral blood and bone marrow (BM) of healthy individuals in China by using 2 panel 4-color flow cytometry (FCM). The percentage and absolute number of Lin-HLA-DR+CD11chiBDCA1+ myeloid DC (mDC) and Lin-HLA-DR+CD123hiBDCA2+ plasmacytoid DC (pDC) were detected in 35 normal BM (NBM) and 29 normal peripheral blood (NPB), the results were compared with the Lin-HLA-DR+CD11chimDC and Lin-HLA-DR+CD123hi pDC obtained by 3-color FCM. The results indicated that both absolute count of DC subset and relative count of pDC in BM were decreased along with increase of age, the absolute count of DC subset in male BM was higher than that in femoral BM (p<0.05). The DC subsets in NBM and NPB were different whatever by 3 or 4-color cytometric analysis, there were more mDCs than pDCs in PB and the ratio of mDC to pDC was 2.70 and 2.31 respectively. In contrast, pDCs predominated in BM, the ratio of mDC to pDC in BM was 0.90 and 0.71 respectively. The quantity of DC subsets significantly correlated to both frequency (mDC r=0.86; pDC r=0.96, p<0.05) and absolute number (mDC r=0.95; pDC r=0.98, p<0.05) between 3 and 4-color cytometric analysis. The quantity of DC subsets in PB and BM were significantly different, counted by 3 and 4-color cytometric analysis except pDC in PB (p<0.001). The quantity of DC subsets were much higher by 3-color than that by 4-color analysis. Since some Lin-HLA-DR+CD11chimDC and Lin-HLA-DR+CD123hi pDC were BDCA1- and BDCA-2dim/- respectively, that more were in BM than in PB. It is concluded that the DC absolute enumeration is correlated with sample type, gender, age and total nucleated cells (p<0.05). 4-color antibody combination may help to identify the real DC subsets in BM. DC subsets in NBM and NPB are different, that more mDC are in PB whereas more pDC in BM.