Anfray Clément, Ummarino Aldo, Calvo Alfonso, Allavena Paola, Torres Andón Fernando
Department of Immunology, Humanitas Clinical and Research Center-IRCCS, Rozzano-Milano, Italy.
Humanitas University, Rozzano-Milano, Italy.
Methods Mol Biol. 2023;2614:93-108. doi: 10.1007/978-1-0716-2914-7_7.
In most solid cancers, tumor-associated macrophages (TAMs) infiltrating the tumor microenvironment (TME) represent a major population of immunosuppressive cells. This correlates with poor prognosis and resistance to antitumoral therapies, including immune checkpoint inhibitors. Although initial preclinical studies were primarily meant to deplete macrophages in the TME or prevent their recruitment at tumor sites, recent evidence has indicated that the reprogramming of macrophages into cytotoxic effectors might be more beneficial in eliciting an effective antitumor immune response. Taking this into consideration, the comprehensive analysis of the phenotype and function of macrophages in the TME, and their interaction with cancer cells or other immune cells, has become of paramount importance in oncological research. Accordingly, here we explain the experimental procedures for the in vivo evaluation of tumor progression and response to therapy, with a particular focus on the detailed analysis of TAMs and related immune cells in the TME by flow cytometry, RNA analysis, and multiplex immunophenotyping. The output generated through these experiments allow researchers to test the efficacy of new therapeutic strategies on targeting.
在大多数实体癌中，浸润肿瘤微环境（TME）的肿瘤相关巨噬细胞（TAM）是免疫抑制细胞的主要群体。这与预后不良以及对抗肿瘤疗法（包括免疫检查点抑制剂）的耐药性相关。尽管最初的临床前研究主要旨在耗尽TME中的巨噬细胞或阻止它们在肿瘤部位的募集，但最近的证据表明，将巨噬细胞重编程为细胞毒性效应细胞可能在引发有效的抗肿瘤免疫反应方面更有益。考虑到这一点，对TME中巨噬细胞的表型和功能及其与癌细胞或其他免疫细胞的相互作用进行全面分析，在肿瘤学研究中变得至关重要。因此，在这里我们解释了体内评估肿瘤进展和对治疗反应的实验程序，特别关注通过流式细胞术、RNA分析和多重免疫表型分析对TME中的TAM和相关免疫细胞进行详细分析。通过这些实验产生的结果使研究人员能够测试新治疗策略在靶向方面的疗效。
