使用流式细胞术分析小鼠结肠癌肿瘤相关髓系来源抑制细胞中精氨酸酶 I 表达的方案。

Protocol for analyzing arginase I expression in tumor-associated myeloid-derived suppressor cells from murine colon cancer using flow cytometry.

机构信息

Department of Translational Hematology and Oncology Research, Cleveland Clinic Foundation, 9500 Euclid Avenue, Cleveland, OH, USA.

Department of Translational Hematology and Oncology Research, Cleveland Clinic Foundation, 9500 Euclid Avenue, Cleveland, OH, USA; Case Comprehensive Cancer Center, Cleveland, OH, USA.

出版信息

STAR Protoc. 2024 Sep 20;5(3):103222. doi: 10.1016/j.xpro.2024.103222. Epub 2024 Jul 31.

DOI:10.1016/j.xpro.2024.103222

PMID:39088325

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11342165/

Abstract

Arginase1 (ARG1) is a metabolic enzyme that is highly expressed in tumor-associated myeloid-derived suppressor cells (MDSCs) and causes the dysfunction of tumor-reactive T cells. Here, we present a protocol for detecting ARG1 expression in tumor MDSCs from a murine model of colon cancer using flow cytometry. We describe steps for tumor tissue processing, antibody staining, and data acquisition. We then detail procedures for identifying MDSC subsets and detecting ARG1 expression using a precise gating strategy. For complete details on the use and execution of this protocol, please refer to Zhang et al..

摘要

精氨酸酶 1（ARG1）是一种代谢酶，在肿瘤相关髓系来源的抑制性细胞（MDSCs）中高度表达，并导致肿瘤反应性 T 细胞功能障碍。在这里，我们提供了一种使用流式细胞术检测结肠癌小鼠模型中肿瘤 MDSC 中 ARG1 表达的方案。我们描述了肿瘤组织处理、抗体染色和数据采集的步骤。然后，我们详细介绍了使用精确门控策略鉴定 MDSC 亚群和检测 ARG1 表达的步骤。有关此方案的使用和执行的完整详细信息，请参阅 Zhang 等人的研究。