Monitoring dendritic cells in clinical practice using a new whole blood single-platform TruCOUNT assay
Monitoring dendritic cells in clinical practice using a new whole blood single-platform TruCOUNT assay
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DOI：
10.1016/j.jim.2003.10.006
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发表时间：
2004-01-01
影响因子：
2.2
通讯作者：
Hart, DNJ
中科院分区：
文献类型：
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作者：
Vuckovic, S;Gardiner, D;Hart, DNJ
来自不同 DC 亚群的树突状细胞 (DC) 是正常人类免疫反应的重要贡献者。尽管如此，能够精确计数 DC 的可靠测定方法的发展进展缓慢。我们现已开发出一种基于 TruCOUN™ 微珠和全血“Lyse/No-Wash”方案的新型单平台流式细胞术检测，可对 CD14(-) 血液 DC 亚群进行精确计数：CD11c(+)CD16(-) DC、CD11c(+)CD16(+) DC、CD123(hi) DC、CD1c(+) DC 和 BDCA-3(+) DC。该测定需要50μl全血；不依赖血液分析仪来获得绝对 DC 计数；允许在收集后对 EDTA 样品 24 It 进行 DC 计数；适用于脐带血和外周血。数据具有高度可重复性，测定内和测定间变异系数分别小于 3% 和 11%。该测定不会产生 DC-T 淋巴细胞结合物，而这种结合物会导致传统梯度密度分离程序中的 DC 计数异常。使用 TruCOUNT 检测，我们确定健康个体的绝对血液 DC 计数随着年龄的增长而减少。在初步研究中，我们发现与年龄匹配的对照相比，III/IV期乳腺癌患者的绝对血液CD11c(+)CD16(+) DC计数显着低于I/II期乳腺癌患者，并且多发性骨髓瘤患者的绝对血液CD123(hi) DC计数较低。这些数据表明 DC 计数技术的科学进步将导致 DC 计数的全球标准化，并获得有临床意义的数据。 (C) 2003 Elsevier B.V. 保留所有权利。
Dendritic cells (DC) from distinct DC subsets are essential contributors to normal human immune responses. Despite this, reliable assays that enable DC to be counted precisely have been slow to evolve. We have now developed a new single-platform flow cytometric assay based on TruCOUN(TM) beads and the whole blood "Lyse/No-Wash" protocol that allows precise counting of the CD14(-) blood DC subsets: CD11c(+)CD16(-) DC, CD11c(+)CD16(+) DC, CD123(hi) DC, CD1c(+) DC and BDCA-3(+) DC. This assay requires 50 mul of whole blood; does not rely on a hematology blood analyser for the absolute DC counts; allows DC counting in EDTA samples 24 It after collection; and is suitable for cord blood and peripheral blood. The data is highly reproducible with intra-assay and inter-assay coefficients of variation less than 3% and 11%, respectively. This assay does not produce the DC-T lymphocyte conjugates that result in DC counting abnormalities in conventional gradient-density separation procedures. Using the TruCOUNT assay, we established that absolute blood DC counts reduce with age in healthy individuals. In preliminary studies, we found a significantly lower absolute blood CD11c(+)CD16(+) DC count in stage III/IV versus stage I/II breast carcinoma patients and a lower absolute blood CD123(hi) DC count in multiple myeloma patients, compared to age-matched controls. These data indicate that scientific progress in DC counting technology will lead to the global standardization of DC counting and allow clinically meaningful data to be obtained. (C) 2003 Elsevier B.V. All rights reserved.