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A guide to interrogating immunometabolism

Nature Reviews Immunology volume 21, pages 637–652 (2021)Cite this article

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Abstract

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.

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Fig. 1: A general immunometabolism workflow.
Fig. 2: A guide to using extracellular flux assays for glycolysis.
Fig. 3: A guide to examination of oxidative phosphorylation and mitochondrial function by extracellular flux assays.
Fig. 4: Metabolomics workflow.

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Acknowledgements

The authors thank C. Deeter and K. V. Tormos at Agilent for helpful resources and discussion.

Funding

This work was supported by T32 DK101003 (K.V.), T32 DK094775 (H.S.H), K00 CA234920 (J.E.B.), T32 GM007347 (A.S.), 1R37CA237421, R01CA248160, R01CA244931 (C.A.L.) and R01 CA217987 and R01 DK105550 (J.C.R.).

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Author notes

  1. These authors contributed equally: Kelsey Voss, Hanna S. Hong.

Authors and Affiliations

  1. Vanderbilt Center for Immunobiology, Department of Pathology, Microbiology, and Immunology, Vanderbilt University Medical Center, Nashville, TN, USA

    Kelsey Voss, Jackie E. Bader, Ayaka Sugiura & Jeffrey C. Rathmell

  2. Rogel Cancer Center, University of Michigan, Ann Arbor, MI, USA

    Hanna S. Hong & Costas A. Lyssiotis

  3. Department of Molecular and Integrative Physiology, University of Michigan, Ann Arbor, MI, USA

    Hanna S. Hong & Costas A. Lyssiotis

  4. Department of Internal Medicine, Division of Gastroenterology and Hepatology, University of Michigan, Ann Arbor, MI, USA

    Costas A. Lyssiotis

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Contributions

K.V. conceptualized the review and wrote the first draft. H.S.H. wrote the metabolomics section and contributed considerably to revisions. J.E.B. and A.S. contributed intellectually to the review, wrote some portions of the review and assisted with edits. C.A.L. and J.C.R. supervised and edited the manuscript.

Corresponding author

Correspondence to Jeffrey C. Rathmell.

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Competing interests

J.C.R. holds stock equity in Sitryx and within the past 2 years has received unrelated research support, travel and honoraria from Incyte, Sitryx, Caribou, Nirogy, Kadmon, Calithera, Tempest, Merck, Mitobridge and Pfizer. The other authors declare no competing interests.

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Nature Reviews Immunology thanks N. Chandel and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.

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Glossary

Microbiota–gut–brain axis

The network that enables bidirectional communication between gut bacteria and the brain.

Electrochemical gradient

A gradient of electrochemical potential; in the case of the mitochondrion, to enable protons to move across the inner mitochondrial membrane.

Anaplerotic pathway

Metabolic pathway, the activity of which replenishes pools of intermediates of the tricarboxylic acid cycle, which can also serve as precursors for other anabolic processes.

Fragmentation state

The status of elongated or fused mitochondria versus smaller mitochondria as a result of fission.

Metabolons

Non-covalent complexes of metabolic enzymes in a metabolic pathway, resulting in increased spatiotemporal efficiency.

Metabolome

The complete set of metabolites present in a cell, biological fluid or tissue sample.

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Voss, K., Hong, H.S., Bader, J.E. et al. A guide to interrogating immunometabolism. Nat Rev Immunol 21, 637–652 (2021). https://doi.org/10.1038/s41577-021-00529-8

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