Life Science Institute, Department of Microbiology and Immunology, University of British Columbia, Vancouver, BC, Canada.
Methods Mol Biol. 2022;2508:147-168. doi: 10.1007/978-1-0716-2376-3_12.
Myeloid cells are a highly heterogeneous group of innate immune cells which include a diverse collection of cell types and cell states. Distinct subsets can impact tumor progression differently, with conventional type 1 DCs important in protective anti-tumor immune responses, while immunosuppressive tumor-associated macrophages and myeloid-derived suppressive cells (MDSCs) play tumor-promoting roles. Deep phenotyping of myeloid cells using single-cell technologies such as mass cytometry provides the unprecedented opportunity to comprehensively characterize the underlying heterogeneity of myeloid cells. Here we provide a detailed end-to-end workflow including both experimental and computational protocols enabling deep phenotyping of tumor-infiltrating myeloid cells using mass cytometry. A protocol that facilitates interrogation of phosphoproteins in circulating and tumor-infiltrating myeloid cells has been provided together with detailed scripts for Phenograph analysis of tumor-infiltrating myeloid cells.
髓样细胞是一组高度异质性的先天免疫细胞，包括多种不同的细胞类型和细胞状态。不同的亚群可能会以不同的方式影响肿瘤的进展，传统的 1 型树突状细胞（DC）在保护性抗肿瘤免疫反应中很重要，而免疫抑制性肿瘤相关巨噬细胞和髓系来源的抑制性细胞（MDSCs）则发挥促进肿瘤的作用。使用单细胞技术（如质谱流式细胞术）对髓样细胞进行深度表型分析，提供了全面描述髓样细胞潜在异质性的前所未有的机会。在这里，我们提供了一个详细的端到端工作流程，包括实验和计算方案，可使用质谱流式细胞术对肿瘤浸润髓样细胞进行深度表型分析。我们还提供了一种方便检测循环和肿瘤浸润髓样细胞中磷酸化蛋白的方案，并提供了用于分析肿瘤浸润髓样细胞的 Phenograph 详细脚本。
