Maby Pauline, Corneau Aurélien, Galon Jérôme
INSERM, Laboratory of Integrative Cancer Immunology, Équipe Labellisée Ligue Contre le Cancer, Sorbonne Université, Sorbonne Paris Cité, Université Paris Descartes, Université Paris Diderot, Centre de Recherche des Cordeliers, Paris, France.
Sorbonne-Université, UMS037 PASS, Plateforme de Cytometrie CyPS, Faculté des Sciences Sorbonne-Université, Paris, France.
Methods Enzymol. 2020;632:339-368. doi: 10.1016/bs.mie.2019.07.025. Epub 2019 Oct 18.
The anti-tumor immune response plays a crucial role in cancer patient outcome as well as in response to the growing family of immunotherapeutic treatments. Improving patient prognostic and therapeutic management requires a better knowledge of the tumor microenvironment, for which a deep characterization of tumor-infiltrating immune populations is instrumental. Mass cytometry represents an excellent tool in tumor Immunology, as it allows the simultaneous analysis of >40 markers on single cells. In this chapter, we review challenging technical aspects of the mass cytometry phenotyping of tumor infiltrating immune cells, focusing on fresh human solid tumor samples. We first explain how to design mass cytometry experiments, then provide detailed protocols to isolate mononuclear immune cells from solid tissues and to stain them for an acquisition on a mass cytometer. We also discuss how to optimize the preparation of single immune cell samples, and how to ensure the reproducibility of data generated from distinct fresh human samples. Finally, we provide troubleshooting suggestions for difficult sample acquisitions on a mass cytometer.
抗肿瘤免疫反应在癌症患者的预后以及对日益增多的免疫治疗方法的反应中起着至关重要的作用。改善患者的预后和治疗管理需要更好地了解肿瘤微环境，而深入表征肿瘤浸润免疫细胞群体对此至关重要。质谱流式细胞术是肿瘤免疫学中的一种出色工具，因为它能够同时分析单个细胞上的40多种标志物。在本章中，我们回顾了肿瘤浸润免疫细胞质谱流式细胞术表型分析中具有挑战性的技术方面，重点是新鲜的人类实体瘤样本。我们首先解释如何设计质谱流式细胞术实验，然后提供从实体组织中分离单核免疫细胞并对其进行染色以便在质谱流式细胞仪上采集的详细方案。我们还讨论了如何优化单个免疫细胞样本的制备，以及如何确保从不同的新鲜人类样本中生成的数据的可重复性。最后，我们针对在质谱流式细胞仪上进行困难样本采集提供故障排除建议。
