Dynamics of Immune Responses Unit, Institut Pasteur, Université de Paris Cité, Inserm U1223, Paris, France.
Vaccine Research Institute, Creteil, France.
EMBO J. 2022 Oct 4;41(19):e111528. doi: 10.15252/embj.2022111528. Epub 2022 Aug 23.
The regulation of cellular energy metabolism is central to most physiological and pathophysiological processes. However, most current methods have limited ability to functionally probe metabolic pathways in individual cells. Here, we describe SPICE-Met (Single-cell Profiling and Imaging of Cell Energy Metabolism), a method for profiling energy metabolism in single cells using flow cytometry or imaging. We generated a transgenic mouse expressing PercevalHR, a fluorescent reporter for cellular ATP:ADP ratio. Modulation of PercevalHR fluorescence with metabolic inhibitors was used to infer the dependence of energy metabolism on oxidative phosphorylation and glycolysis in defined cell populations identified by flow cytometry. We applied SPICE-Met to analyze T-cell memory development during vaccination. Finally, we used SPICE-Met in combination with real-time imaging to dissect the heterogeneity and plasticity of energy metabolism in single macrophages ex vivo and identify three distinct metabolic patterns. Functional probing of energy metabolism with single-cell resolution should greatly facilitate the study of immunometabolism at a steady state, during disease pathogenesis or in response to therapy.
细胞能量代谢的调节是大多数生理和病理生理过程的核心。然而，大多数现有的方法在功能上探测单个细胞中的代谢途径的能力有限。在这里，我们描述了 SPICE-Met（单细胞能量代谢的分析和成像），这是一种使用流式细胞术或成像技术在单细胞中进行能量代谢分析的方法。我们生成了一种表达 PercevalHR 的转基因小鼠，PercevalHR 是一种用于细胞内 ATP:ADP 比率的荧光报告蛋白。通过代谢抑制剂对 PercevalHR 荧光的调节，来推断在通过流式细胞术鉴定的特定细胞群中，能量代谢对氧化磷酸化和糖酵解的依赖性。我们应用 SPICE-Met 来分析疫苗接种过程中 T 细胞记忆的发展。最后，我们使用 SPICE-Met 与实时成像相结合，在体外分离单个巨噬细胞中能量代谢的异质性和可塑性，并确定三种不同的代谢模式。通过单细胞分辨率进行能量代谢的功能探测，应该会极大地促进稳态、疾病发病机制或治疗反应期间的免疫代谢研究。
