Abstract

The ketone bodies β-hydroxybutyrate (BHB) and acetoacetate (AcAc) support mammalian survival during states of energy deficit by serving as alternative sources of ATP1. BHB levels are elevated by starvation, caloric restriction, high-intensity exercise, or the low-carbohydrate ketogenic diet2. Prolonged fasting reduces inflammation; however, the impact that ketones and other alternative metabolic fuels produced during energy deficits have on the innate immune response is unknown2,3,4,5,6. We report that BHB, but neither AcAc nor the structurally related short-chain fatty acids butyrate and acetate, suppresses activation of the NLRP3 inflammasome in response to urate crystals, ATP and lipotoxic fatty acids. BHB did not inhibit caspase-1 activation in response to pathogens that activate the NLR family, CARD domain containing 4 (NLRC4) or absent in melanoma 2 (AIM2) inflammasome and did not affect non-canonical caspase-11, inflammasome activation. Mechanistically, BHB inhibits the NLRP3 inflammasome by preventing K+ efflux and reducing ASC oligomerization and speck formation. The inhibitory effects of BHB on NLRP3 are not dependent on chirality or starvation-regulated mechanisms like AMP-activated protein kinase (AMPK), reactive oxygen species (ROS), autophagy or glycolytic inhibition. BHB blocks the NLRP3 inflammasome without undergoing oxidation in the TCA cycle, and independently of uncoupling protein-2 (UCP2), sirtuin-2 (SIRT2), the G protein–coupled receptor GPR109A or hydrocaboxylic acid receptor 2 (HCAR2). BHB reduces NLRP3 inflammasome–mediated interleukin (IL)-1β and IL-18 production in human monocytes. In vivo, BHB or a ketogenic diet attenuates caspase-1 activation and IL-1β secretion in mouse models of NLRP3-mediated diseases such as Muckle–Wells syndrome, familial cold autoinflammatory syndrome and urate crystal–induced peritonitis. Our findings suggest that the anti-inflammatory effects of caloric restriction or ketogenic diets may be linked to BHB-mediated inhibition of the NLRP3 inflammasome.

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Acknowledgements

We thank M. Koch, Y. Kui, P. Chang and D. Albarado for technical assistance, and V. M. Dixit (Genentech) and R. Medzhitov (Yale School of Medicine) for helpful discussions and for providing knockout mice. Salmonella typhimurium and Francisella tularensis (U112) were provided by D. Monack (Stanford University School of Medicine) and J. Teale (University of Texas at San Antonio). M.B. and A.B. were supported by the National Institute on Aging–Intramural Research Program. D.D'A. was supported by the Office of Naval Research (ONR) Grant N000141310062. V.D.D. was supported in part by the grants from National Institutes of Health (AG043608, AG31797, DK090556 and AI105097).

Author information

Author notes

    • Yun-Hee Youm
    •  & Kim Y Nguyen

    These authors contributed equally to this work.

Affiliations

  1. Section of Comparative Medicine and Program on Integrative Cell Signaling and Neurobiology of Metabolism, Yale School of Medicine, New Haven, Connecticut, USA.

    • Yun-Hee Youm
    • , Kim Y Nguyen
    • , Emily L Goldberg
    • , Tamas L Horvath
    •  & Vishwa Deep Dixit
  2. Department of Nutrition Sciences, Purdue University, West Lafayette, Indiana.

    • Ryan W Grant
  3. Laboratory of Molecular Biology and Immunology, National Institute on Aging, National Institutes of Health (NIH), Baltimore, Maryland, USA.

    • Monica Bodogai
    •  & Arya Biragyn
  4. Department of Biomedical Engineering, Yale University, New Haven, Connecticut, USA.

    • Dongin Kim
    •  & Tarek M Fahmy
  5. Department of Molecular Pharmacology and Physiology, University of South Florida, Tampa, Florida, USA.

    • Dominic D'Agostino
  6. Department of Geology and Geophysics, Yale University, New Haven, Connecticut, USA.

    • Noah Planavsky
  7. Department of Immunology, St. Jude Children's Hospital, Memphis, Tennessee, USA.

    • Christopher Lupfer
    •  & Thirumala D Kanneganti
  8. Department of Biochemistry and Molecular Biology, Thomas Jefferson University, Philadelphia, Pennsylvania, USA.

    • Seokwon Kang
    •  & Emad Alnemri
  9. Diabetes and Obesity Research Center, Sanford-Burnham Medical Research Institute, Orlando, Florida, USA.

    • Peter A Crawford
  10. Department of Immunobiology, Yale School of Medicine, New Haven, Connecticut, USA.

    • Vishwa Deep Dixit

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Contributions

Y. H.Y. and K.Y.N. designed and conducted the majority of in vitro and all in vivo experiments, analyzed and interpreted the data, and participated in writing the manuscript. R.W.G. participated in design and conduct of inflammasome activation experiments. E.L.G. performed ASC speck and neutrophil assays. M.B. and A.B. performed the human monocytes experiments. D.K. and T.M.F. synthesized the BHB–nanolipogels and conducted control experiments to determine the dose response. D.D'A. formulated the ketone diester diet. N.P. conducted the ICP-MS experiments to determine K+ efflux. C.L. and T.D.K. conducted the F. tularensis and S. typhimurium infection experiments. T.L.H. designed the experiments and provided essential reagents for experiments involving mitochondrial ROS and UCP2. P.A.C. generated the macrophage-specific, Scot-deficient mice and contributed to experiment design. S.K. and E.A. designed and conducted the ASC oligomerization experiments. V.D.D. conceived and supervised the project, interpreted the data, and wrote the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Vishwa Deep Dixit.

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DOI

https://doi.org/10.1038/nm.3804

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