Department of Immunology, Duke University, Durham, NC 27710, USA.
Department of Molecular Genetics and Microbiology, Duke University, Durham, NC 27710, USA.
STAR Protoc. 2020 Dec 30;2(1):100244. doi: 10.1016/j.xpro.2020.100244. eCollection 2021 Mar 19.
Characterizing cytokine production is important for properly understanding immunologic responses. Cytokine reporter mice are limited by the need to cross markers into various knockout backgrounds and by availability of reporters of interest. To overcome this, we utilize injection of brefeldin A into mice to enable flow cytometric analysis of cytokine production during a bacterial infection. While we evaluate IFN-γ production during infection, this protocol can be applied to other cytokines and other mouse models. For complete details on the use and execution of this protocol, please refer to Kovacs et al. (2020) and Liu and Whitton (2005).
对细胞因子产生进行特征分析对于正确理解免疫反应非常重要。细胞因子报告小鼠受到需要将标记物交叉引入各种基因敲除背景以及可用的感兴趣报告物的限制。为了克服这一问题，我们利用向小鼠注射布雷菲德菌素 A 来实现对细菌感染期间细胞因子产生的流式细胞分析。虽然我们在感染过程中评估 IFN-γ 的产生，但该方案可应用于其他细胞因子和其他小鼠模型。有关该方案使用和实施的完整详细信息，请参考 Kovacs 等人（2020 年）和 Liu 和 Whitton（2005 年）。
