The metabolic charts memorized in early biochemistry courses, and then later forgotten, have come back to haunt many immunologists with new recognition of the importance of these pathways. Metabolites and the activity of metabolic pathways drive energy production, macromolecule synthesis, intracellular signalling, post-translational modifications and cell survival. Immunologists who identify a metabolic phenotype in their system are often left wondering where to begin and what does it mean? Here, we provide a framework for navigating and selecting the appropriate biochemical techniques to explore immunometabolism. We offer recommendations for initial approaches to develop and test metabolic hypotheses and how to avoid common mistakes. We then discuss how to take things to the next level with metabolomic approaches, such as isotope tracing and genetic approaches. By proposing strategies and evaluating the strengths and weaknesses of different methodologies, we aim to provide insight, note important considerations and discuss ways to avoid common misconceptions. Furthermore, we highlight recent studies demonstrating the power of these metabolic approaches to uncover the role of metabolism in immunology. By following the framework in this Review, neophytes and seasoned investigators alike can venture into the emerging realm of cellular metabolism and immunity with confidence and rigour.

早期生物化学课程中记住的代谢图表，后来又被遗忘，却再次困扰着许多免疫学家，让他们对这些途径的重要性有了新的认识。代谢物和代谢途径的活性驱动能量产生、大分子合成、细胞内信号传导、翻译后修饰和细胞存活。免疫学家在识别系统中的代谢表型时常常想知道从哪里开始以及它意味着什么？在这里，我们提供了一个框架，用于导航和选择适当的生化技术来探索免疫代谢。我们为开发和测试代谢假设的初步方法以及如何避免常见错误提供建议。然后我们讨论如何通过代谢组学方法（例如同位素追踪和遗传方法）将事情提升到一个新的水平。通过提出策略并评估不同方法的优缺点，我们旨在提供见解、指出重要的考虑因素并讨论避免常见误解的方法。此外，我们重点介绍了最近的研究，这些研究证明了这些代谢方法在揭示代谢在免疫学中的作用方面的力量。通过遵循本综述中的框架，新手和经验丰富的研究人员都可以充满信心和严谨地进入细胞代谢和免疫的新兴领域。