Dendritic Cell Research, ANZAC Research Institute, Sydney, NSW, Australia; Sydney Medical School, University of Sydney, Sydney, NSW, Australia.
Dendritic Cell Research, ANZAC Research Institute , Sydney, NSW, Australia.
Clin Transl Immunology. 2016 Apr 8;5(4):e68. doi: 10.1038/cti.2016.5. eCollection 2016 Apr.
HLDA10 collated a panel of monoclonal antibodies (mAbs) that primarily recognised molecules on human myeloid cell and dendritic cell (DC) populations. As part of the studies, we validated a backbone of mAbs to delineate monocyte and DC populations from peripheral blood. The mAb backbone allowed identification of monocyte and DC subsets using fluorochromes that were compatible with most 'off the shelf' or routine flow cytometers. Three laboratories used this mAb backbone to assess the HLDA10 panel on blood monocytes and DCs. Each laboratory was provided with enough mAbs to perform five repeat experiments. The data were collated and analysed using Spanning-tree Progression Analysis of Density-normalised Events (SPADE). The data were interrogated for inter- and intra-laboratory variability. The results highlight the definition of DC populations using current readily available reagents. This collaborative process provides the broader scientific community with an invaluable data set that validates mAbs to leucocyte surface molecules.
HLDA10 汇集了一组单克隆抗体 (mAbs),这些抗体主要识别人类髓样细胞和树突状细胞 (DC) 群体上的分子。作为研究的一部分,我们验证了一组 mAbs 来区分外周血中的单核细胞和 DC 群体。mAb 骨干可以使用与大多数“现成”或常规流式细胞仪兼容的荧光染料来识别单核细胞和 DC 亚群。三个实验室使用这种 mAb 骨干来评估血液单核细胞和 DC 上的 HLDA10 小组。每个实验室都提供了足够的 mAbs 来进行五次重复实验。使用密度归一化事件的扩展树状进步分析 (SPADE) 对数据进行整理和分析。对实验室间和实验室内的变异性进行了调查。结果强调了使用当前现成试剂定义 DC 群体。这个协作过程为更广泛的科学界提供了一个非常宝贵的数据集,验证了针对白细胞表面分子的 mAbs。