Osmium-Labeled Microspheres for Bead-Based Assays in Mass Cytometry
Osmium-Labeled Microspheres for Bead-Based Assays in Mass Cytometry
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DOI：
10.4049/jimmunol.1801640
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发表时间：
2019-05-15
影响因子：
4.4
通讯作者：
Mei, Henrik E.
中科院分区：
文献类型：
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作者：
Budzinski, Lisa;Schulz, Axel R.;Mei, Henrik E.
聚苯乙烯微球在流式细胞术中有着广泛的应用。在质量细胞术中实施基于头部的测定是需要的，但由于缺乏检测所需的元素标签而受到阻碍。在这项研究中，我们引入了稳定的四氧化锇标记作为一种通用的方法，用于生成易于通过细胞计数术检测的功能化珠子。我们展示了锇标记珠在细胞计数中用于信号溢出补偿的效用，并且，引人注目的是，它们在定量抗体结合能力测定中的应用结合了人类PBMC的高维谱，使得系统评估了大量细胞表型的受体表达谱。该分析证实，活动性系统性红斑狼疮患者单核细胞siglec1表达增加，此外，通过调节性和记忆性CD4 T细胞和髓系树突状细胞的HLA-DR表达，显示CD4表达的相关降低，指出免疫突触的缺陷串音可能限制系统性红斑狼疮的免疫反应。通过将传统的流式细胞仪珠转化为适用于质量细胞仪的珠，我们的方法为生物医学研究中通过质量细胞仪在高维细胞谱研究中广泛实施基于珠的检测铺平了道路。
Polystyrene beads are broadly applied in flow cytometry. Implementing bead-based assays in mass cytometry is desired but hampered by the lack of an elemental label required for their detection. In this study, we introduce stable osmium tetroxide labeling as a universal approach for generating functionalized beads readily detectable by mass cytometry. We demonstrate the utility of osmium-labeled beads for signal spillover compensation in mass cytometry, and, strikingly, their application in quantitative Ab-binding capacity assays combined with high-dimensional profiling of human PBMC enabled the systematic assessment of receptor expression profiles across large numbers of cellular phenotypes. This analysis confirmed increased monocytic Siglec-1 expression in active systemic lupus erythematosus patients and, additionally, revealed interrelated reductions of CD4 expression by regulatory and memory CD4 T cells and HLA-DR expression by myeloid dendritic cells, pointing toward defective cross-talk at the immunological synapse that may limit immune responses in systemic lupus erythematosus. By converting conventional flow cytometry beads into beads suitable for mass cytometry, our approach paves the way toward the broad implementation of bead-based assays in high-dimensional cell profiling studies by mass cytometry in biomedical research.