Institute of Molecular Life Sciences, University of Zürich, Winterthurerstrasse 190, 8057, Zürich, Switzerland.
Department of Biology, Institute of Molecular System Biology, Swiss Federal Institute of Technology (ETH Zürich), Otto-Stern-Weg 3, Zürich, Switzerland.
Sci Rep. 2019 Feb 13;9(1):1925. doi: 10.1038/s41598-018-38127-9.
Phagocytosis is a process in which target cells or particles are engulfed and taken up by other cells, typically professional phagocytes; this process is crucial in many physiological processes and disease states. The detection of targets for phagocytosis is directed by a complex repertoire of cell surface receptors. Pattern recognition receptors directly detect targets for binding and uptake, while opsonic and complement receptors detect objects coated by soluble factors. However, the importance of single and combinatorial surface marker expression across different phenotypes of professional phagocytes is not known. Here we developed a novel mass cytometry-based phagocytosis assay that enables the simultaneous detection of phagocytic events in combination with up to 40 other protein markers. We applied this assay to distinct monocyte derived macrophage (MDM) populations and found that prototypic M2-like MDMs phagocytose more E. coli than M1-like MDMs. Surface markers such as CD14, CD206, and CD163 rendered macrophages phagocytosis competent, but only CD209 directly correlated with the amount of particle uptake. Similarly, M2-like MDMs also phagocytosed more cancer cells than M1-like MDMs but, unlike M1-like MDMs, were insensitive to anti-CD47 opsonization. Our approach facilitates the simultaneous study of single-cell phenotypes, phagocytic activity, signaling and transcriptional events in complex cell mixtures.
吞噬作用是一种将靶细胞或颗粒被其他细胞吞噬并摄取的过程，通常是专业的吞噬细胞；这个过程在许多生理过程和疾病状态中都至关重要。吞噬作用的靶标的检测是由细胞表面受体的复杂组合来指导的。模式识别受体直接检测结合和摄取的靶标，而调理素和补体受体检测被可溶性因子包裹的物体。然而，不同表型的专业吞噬细胞中单个和组合的表面标记表达的重要性尚不清楚。在这里，我们开发了一种基于液质联用技术的新的吞噬作用测定法，该方法能够同时检测吞噬作用事件，并与多达 40 种其他蛋白质标记物结合检测。我们将该测定法应用于不同的单核细胞衍生的巨噬细胞（MDM）群体，并发现典型的 M2 样 MDM 比 M1 样 MDM 吞噬更多的大肠杆菌。表面标记物如 CD14、CD206 和 CD163 使巨噬细胞具有吞噬能力，但只有 CD209 与颗粒摄取量直接相关。同样，M2 样 MDM 也比 M1 样 MDM 吞噬更多的癌细胞，但与 M1 样 MDM 不同，它们对抗 CD47 调理素作用不敏感。我们的方法促进了在复杂细胞混合物中同时研究单细胞表型、吞噬活性、信号和转录事件。
