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Neutrophil extracellular traps enriched in oxidized mitochondrial DNA are interferogenic and contribute to lupus-like disease

Nature Medicine volume 22, pages 146–153 (2016)Cite this article

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Abstract

Neutrophil extracellular traps (NETs) are implicated in autoimmunity, but how they are generated and their roles in sterile inflammation remain unclear. Ribonucleoprotein immune complexes (RNP ICs), inducers of NETosis, require mitochondrial reactive oxygen species (ROS) for maximal NET stimulation. After RNP IC stimulation of neutrophils, mitochondria become hypopolarized and translocate to the cell surface. Extracellular release of oxidized mitochondrial DNA is proinflammatory in vitro, and when this DNA is injected into mice, it stimulates type I interferon (IFN) signaling through a pathway dependent on the DNA sensor STING. Mitochondrial ROS are also necessary for spontaneous NETosis of low-density granulocytes from individuals with systemic lupus erythematosus. This was also observed in individuals with chronic granulomatous disease, who lack NADPH oxidase activity but still develop autoimmunity and type I IFN signatures. Mitochondrial ROS inhibition in vivo reduces disease severity and type I IFN responses in a mouse model of lupus. Together, these findings highlight a role for mitochondria in the generation not only of NETs but also of pro-inflammatory oxidized mitochondrial DNA in autoimmune diseases.

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Figure 1: Mitochondrial ROS support RNP IC–mediated NETosis.
Figure 2: RNP ICs induce mitochondria mobilization and release of oxidized DNA.
Figure 3: Oxidized DNA enhances the inflammatory response in a STING-dependent manner.
Figure 4: SLE LDGs release oxidized mtDNA in a mitochondrial superoxide-dependent manner.
Figure 5: LDGs from individuals with CGD release NETs in a mitochondrial superoxide-dependent manner.
Figure 6: In vivo administration of a mito-ROS scavenger attenuates lupus-like disease in mice.

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Acknowledgements

We thank X. Sun, L. Tanaka (both at University of Washington), and E. Moore (National Institute of Arthritis and Musculoskeletal and Skin Diseases; NIAMS) for technical assistance; W. Wang (University of Washington) for providing mouse mitochondria isolated from hearts; D. Kuhns (NIAID) for scientific input, and R. Siegel (NIAMS) for critical review of the manuscript. We also thank H.-W. Sun, M. Ward (both at NIAMS) and C. Spiekerman (University of Washington) for expert statistical advice. We also thank M. Gale for providing Tmem173-knockout mice. The study was supported by grants from the Washington Research Foundation, Leap for Lupus (K.B.E.), the Wenner-Gren Foundation, the foundation BLANCEFLOR Boncompagni-Ludovisi née Bildt (C.L.), and the Intramural Research Program at NIAMS/NIH (L.P.B., M.P., C.K.S., C.C.-R. and M.J.K.). The funding bodies had no part in the study design; the collection, analysis and interpretation of the data; the writing of the manuscript or the submission.

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  1. Christian Lood and Luz P Blanco: These authors contributed equally to this work.

  2. Keith B Elkon and Mariana J Kaplan: These authors jointly directed this work.

Authors and Affiliations

  1. Division of Rheumatology, University of Washington, Seattle, Washington, USA

    Christian Lood & Keith B Elkon

  2. Systemic Autoimmunity Branch, National Institute of Arthritis and Musculoskeletal and Skin Diseases, National Institutes of Health (NIH), Bethesda, Maryland, USA

    Luz P Blanco, Monica M Purmalek, Carmelo Carmona-Rivera, Carolyne K Smith & Mariana J Kaplan

  3. Laboratory of Host Defenses, National Institute of Allergy and Infectious Diseases, NIH, Bethesda, Maryland, USA

    Suk S De Ravin & Harry L Malech

  4. Department of Immunology, University of Washington, Seattle, Washington, USA

    Jeffrey A Ledbetter & Keith B Elkon

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Contributions

All contributing authors have agreed to the submission of this manuscript for publication. K.B.E. and M.J.K. conceived the study, and analyzed and interpreted results. C.L., L.P.B., M.M.P., C.C.-R. and C.K.S. designed and performed experiments, analyzed data and interpreted results. S.S.D.R. and H.L.M. provided study materials and characterized the CGD cohort. J.A.L. participated in the design and interpretation of the in vitro data, as well as the critical review of the paper. C.L., L.P.B., K.B.E. and M.J.K. wrote the manuscript.

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Correspondence to Keith B Elkon or Mariana J Kaplan.

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Lood, C., Blanco, L., Purmalek, M. et al. Neutrophil extracellular traps enriched in oxidized mitochondrial DNA are interferogenic and contribute to lupus-like disease. Nat Med 22, 146–153 (2016). https://doi.org/10.1038/nm.4027

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