Method for the Analysis of the Tumor Microenvironment by Mass Cytometry: Application to Chronic Lymphocytic Leukemia.
Method for the Analysis of the Tumor Microenvironment by Mass Cytometry: Application to Chronic Lymphocytic Leukemia.
复制标题
肿瘤微环境的细胞计数分析方法在慢性淋巴细胞白血病中的应用。
DOI：
10.3389/fimmu.2020.578176
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发表时间：
2020
影响因子：
7.3
通讯作者：
Largeot A
中科院分区：
文献类型：
--
作者：
Gonder S;Fernandez Botana I;Wierz M;Pagano G;Gargiulo E;Cosma A;Moussay E;Paggetti J;Largeot A
在过去的20年里，对肿瘤微环境（TME）的兴趣呈指数级增长。事实上，现在普遍承认TME在癌症发展、维持、免疫逃逸和对治疗的抵抗中起着至关重要的作用。这也适用于血液恶性肿瘤。相当多的新开发的疗法是针对支持癌症的TME，而不是针对肿瘤细胞本身。然而，TME往往没有明确的定义。此外，每种肿瘤的独特表型和患者之间的变异性限制了此类疗法的成功。最近，我们的团队利用质谱技术在小鼠慢性淋巴细胞白血病（CLL）的背景下揭示了特异性TME。我们发现了两个免疫检查点LAG3和PD1的富集。我们测试了一种基于抗体的免疫疗法，靶向这两种分子。这种抗体组合在治疗鼠CLL中是成功的。在这篇方法文章中，我们提供了一个详细的协议，CLL TME细胞的染色，目的是他们的表征使用质谱细胞术。我们包括面板设计和验证、样品制备和采集、机器设置、质量控制和分析。此外，我们还讨论了这种技术的不同优点和缺陷。
In the past 20 years, the interest for the tumor microenvironment (TME) has exponentially increased. Indeed, it is now commonly admitted that the TME plays a crucial role in cancer development, maintenance, immune escape and resistance to therapy. This stands true for hematological malignancies as well. A considerable amount of newly developed therapies are directed against the cancer-supporting TME instead of targeting tumor cells themselves. However, the TME is often not clearly defined. In addition, the unique phenotype of each tumor and the variability among patients limit the success of such therapies. Recently, our group took advantage of the mass cytometry technology to unveil the specific TME in the context of chronic lymphocytic leukemia (CLL) in mice. We found the enrichment of LAG3 and PD1, two immune checkpoints. We tested an antibody-based immunotherapy, targeting these two molecules. This combination of antibodies was successful in the treatment of murine CLL. In this methods article, we provide a detailed protocol for the staining of CLL TME cells aiming at their characterization using mass cytometry. We include panel design and validation, sample preparation and acquisition, machine set-up, quality control, and analysis. Additionally, we discuss different advantages and pitfalls of this technique.