Ensemble quantitation of absolute metabolite concentrations in T cells reveals conserved features of immunometabolism
T cell diversity and differentiation define individual adaptive immunity. Metabolism fuels adaptive immunity by driving proliferation, differentiation, and activation; however, how individual and lineage differences shape T cell metabolism remains unknown. Here we develop an ensemble method for absolute metabolome quantitation and quantify 84 metabolites in human primary T cells. Liquid chromatography-mass spectrometry of metabolites co-extracted from T cells and 13C-labeled reference cells reveals absolute concentrations en masse. Across subtypes and individuals, T cell metabolomes resemble one another. T cells possess thermodynamically forward-driven glycolysis, high adenylate energy charge, large amino acid pools, and favorable redox ratios for anabolism and antioxidant defense. Across metabolism, metabolite concentrations exceed their associated Michaelis constants and inhibitor constants two thirds and half of the time, respectively. The conserved features of T cell metabolomes underlie a design principle: metabolite levels prime T cells for increased energy demand, oxidative stress, and rapid regulation that accompany immune response.