T cells are integral players in the adaptive immune system that readily adapt their metabolism to meet their energetic and biosynthetic needs. A major hurdle to understand physiologic T-cell metabolism has been the differences between in vitro cell culture conditions and the complex in vivo milieu. To address this, we have developed a protocol that merges traditional immunology infection models with whole-body metabolite infusion and mass-spectrometry-based metabolomic profiling to assess T-cell metabolism in vivo. In this protocol, pathogen-infected mice are infused via the tail vein with an isotopically labeled metabolite (2–6 h), followed by rapid magnetic bead isolation to purify T-cell populations (<1 h) and then stable isotope labeling analysis conducted by mass spectrometry (~1–2 d). This procedure enables researchers to evaluate metabolic substrate utilization into central carbon metabolic pathways (i.e., glycolysis and the tricarboxylic acid cycle) by specific T-cell subpopulations in the context of physiological immune responses in vivo.

T 细胞是适应性免疫系统中不可或缺的参与者，它们很容易适应其新陈代谢以满足其能量和生物合成的需求。了解生理性 T 细胞代谢的一个主要障碍是体外细胞培养条件和复杂的体内环境之间的差异。为了解决这个问题，我们开发了一种方案，将传统的免疫学感染模型与全身代谢物输注和基于质谱的代谢组学分析相结合，以评估体内 T 细胞代谢。在该协议中，病原体感染的小鼠通过尾静脉注入同位素标记的代谢物（2-6 小时），然后快速磁珠分离以纯化 T 细胞群（<1 小时），然后进行稳定同位素标记分析通过质谱（~1-2 d）。