Standardized Single-Platform Assay for Human Monocyte Subpopulations: Lower CD14+CD16++ Monocytes in Females
Standardized Single-Platform Assay for Human Monocyte Subpopulations: Lower CD14+CD16++ Monocytes in Females
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DOI：
10.1002/cyto.a.20942
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发表时间：
2010-09-01
期刊：
影响因子：
3.7
通讯作者：
Zieger-Heitbrock, Loems
中科院分区：
文献类型：
--
作者：
Heimbeck, Irene;Hofer, Thomas P. J.;Zieger-Heitbrock, Loems
我们提出了一种新的单平台测定法，用于测定人血液单核细胞亚群的绝对数量，即，CD 14（++）CD 16（-）和CD 14（+）CD 16（++）单核细胞。针对CD 14、CD 16、CD 45和HLA-DR的四色抗体组合减少了自然杀伤细胞和粒细胞向CD 14（+）CD 16（++）单核细胞门的溢出。对于这些CD 14（+）CD 16（++）单核细胞，试验内变异系数（CV）为4.1%，试验间CV为8.5%。观察40名年龄在18-60岁之间的供体S，我们发现没有年龄依赖性。然而，性别的影响在于，与男性（59.1 +/- 20.3细胞/μ l）相比，女性具有较低的CD 14（+）CD 16（++）单核细胞（45.4 +/- 13.5细胞/μ l）（P < 0.02）。使用这种新的方法，我们可以证实运动会导致CD 14（+）CD 16（++）单核细胞增加三倍以上。此外，我们表明，低剂量的糖皮质激素治疗将耗尽这些细胞。这种稳健的单平台测定可能是监测健康和疾病中单核细胞亚群绝对数量的有用工具。(C)2010年国际细胞计数促进学会
We present a novel single-platform assay for determination of the absolute number of human blood monocyte subpopulations, i.e., the CD14(++)CD16(-) and the CD14(+)CD16(++) monocytes. A four-color combination of antibodies to CD14, CD16, CD45, and HLA-DR reduces the spill-over of natural killer cells and of granulocytes into the CD14(+)CD16(++) monocyte gate. For these CD14(+)CD16(++) monocytes, the intra-assay coefficient of variation (CV) was 4.1% and the inter-assay CV was 8.5%. Looking at a cohort of 40 donor S aged 18-60 years, we found no age dependence. There was however an effect of gender in that females had lower CD14(+)CD16(++) monocytes (45.4 +/- 13.5 cells/mu l) compared with males (59.1 +/- 20.3 cells/mu l) (P < 0.02). Using this novel approach, we can confirm that exercise will lead to more than three-fold increase of the CD14(+)CD16(++) monocytes. Also, we show that therapy with low doses of glucocorticoids will deplete these cells. This robust single-platform assay may be a useful tool for monitoring the absolute number of monocyte subpopulations in health and disease. (C) 2010 International Society for Advancement of Cytometry