Agency for Science, Technology and Research (A*STAR), Singapore Immunology Network (SIgN), Singapore.
Immunoscape Pte. Ltd., Immunos Building, Singapore.
Curr Opin Immunol. 2018 Apr;51:187-196. doi: 10.1016/j.coi.2018.03.023. Epub 2018 Apr 11.
Advancement in methodologies for single cell analysis has historically been a major driver of progress in immunology. Currently, high dimensional flow cytometry, mass cytometry and various forms of single cell sequencing-based analysis methods are being widely adopted to expose the staggering heterogeneity of immune cells in many contexts. Here, we focus on mass cytometry, a form of flow cytometry that allows for simultaneous interrogation of more than 40 different marker molecules, including cytokines and transcription factors, without the need for spectral compensation. We argue that mass cytometry occupies an important niche within the landscape of single-cell analysis platforms that enables the efficient and in-depth study of diverse immune cell subsets with an ability to zoom-in on myeloid and lymphoid compartments in various tissues in health and disease. We further discuss the unique features of mass cytometry that are favorable for combining multiplex peptide-MHC multimer technology and phenotypic characterization of antigen specific T cells. By referring to recent studies revealing the complexities of tumor immune infiltrates, we highlight the particular importance of this technology for studying cancer in the context of cancer immunotherapy. Finally, we provide thoughts on current technical limitations and how we imagine these being overcome.
单细胞分析方法的进展一直是免疫学进步的主要驱动力。目前,高维流式细胞术、质谱流式细胞术和各种基于单细胞测序的分析方法正在被广泛采用,以揭示许多情况下免疫细胞令人惊讶的异质性。在这里,我们重点介绍质谱流式细胞术,这是一种流式细胞术,可以在不进行光谱补偿的情况下同时检测超过 40 种不同的标记分子,包括细胞因子和转录因子。我们认为,质谱流式细胞术在单细胞分析平台的领域中占据着重要的位置,能够高效深入地研究各种免疫细胞亚群,并能够在健康和疾病状态下对各种组织中的髓系和淋巴系进行放大观察。我们进一步讨论了质谱流式细胞术的独特特点,这些特点有利于结合多色肽-MHC 三聚体技术和抗原特异性 T 细胞的表型特征分析。通过参考最近揭示肿瘤免疫浸润复杂性的研究,我们强调了该技术在癌症免疫治疗背景下研究癌症的特别重要性。最后,我们对当前的技术限制以及我们如何想象克服这些限制进行了思考。