Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

Peroxiredoxin family proteins are key initiators of post-ischemic inflammation in the brain

Abstract

Post-ischemic inflammation is an essential step in the progression of brain ischemia-reperfusion injury. However, the mechanism that activates infiltrating macrophages in the ischemic brain remains to be clarified. Here we demonstrate that peroxiredoxin (Prx) family proteins released extracellularly from necrotic brain cells induce expression of inflammatory cytokines including interleukin-23 in macrophages through activation of Toll-like receptor 2 (TLR2) and TLR4, thereby promoting neural cell death, even though intracellular Prxs have been shown to be neuroprotective. The extracellular release of Prxs in the ischemic core occurred 12 h after stroke onset, and neutralization of extracellular Prxs with antibodies suppressed inflammatory cytokine expression and infarct volume growth. In contrast, high mobility group box 1 (HMGB1), a well-known damage-associated molecular pattern molecule, was released before Prx and had a limited role in post-ischemic macrophage activation. We thus propose that extracellular Prxs are previously unknown danger signals in the ischemic brain and that its blocking agents are potent neuroprotective tools.

This is a preview of subscription content, access via your institution

Access options

Rent or buy this article

Prices vary by article type

from$1.95

to$39.95

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Peroxiredoxins in brain lysate are potent inducers of IL-23 expression.
Figure 2: Extracellular peroxiredoxin colocalizes with infiltrating macrophages in the ischemic brain.
Figure 3: Peroxiredoxins induce IL-23 expression via TLR2 and TLR4.
Figure 4: Extracellular release of Prxs contributes to the initiation of post-ischemic inflammation.
Figure 5: Neutralization of extracellular Prxs is neuroprotective within 12 h after stroke onset.
Figure 6: The conserved region of peroxiredoxins was essential for IL-23-inducing activity and the increase in infarct size.

Similar content being viewed by others

References

  1. Moskowitz, M.A., Lo, E.H. & Iadecola, C. The science of stroke: mechanisms in search of treatments. Neuron 67, 181–198 (2010).

    Article  CAS  Google Scholar 

  2. Lo, E.H. Degeneration and repair in central nervous system disease. Nat. Med. 16, 1205–1209 (2010).

    Article  CAS  Google Scholar 

  3. Ooboshi, H. et al. Postischemic gene transfer of interleukin-10 protects against both focal and global brain ischemia. Circulation 111, 913–919 (2005).

    Article  CAS  Google Scholar 

  4. Macrez, R. et al. Stroke and the immune system: from pathophysiology to new therapeutic strategies. Lancet Neurol. 10, 471–480 (2011).

    Article  CAS  Google Scholar 

  5. Iadecola, C. & Anrather, J. The immunology of stroke: from mechanisms to translation. Nat. Med. 17, 796–808 (2011).

    Article  CAS  Google Scholar 

  6. Eltzschig, H.K. & Eckle, T. Ischemia and reperfusion—from mechanism to translation. Nat. Med. 17, 1391–1401 (2011).

    Article  CAS  Google Scholar 

  7. Hurn, P.D. et al. T- and B-cell–deficient mice with experimental stroke have reduced lesion size and inflammation. J. Cereb. Blood Flow Metab. 27, 1798–1805 (2007).

    Article  CAS  Google Scholar 

  8. Liesz, A. et al. Inhibition of lymphocyte trafficking shields the brain against deleterious neuroinflammation after stroke. Brain 134, 704–720 (2011).

    Article  Google Scholar 

  9. Yilmaz, G., Arumugam, T.V., Stokes, K.Y. & Granger, D.N. Role of T lymphocytes and interferon-gamma in ischemic stroke. Circulation 113, 2105–2112 (2006).

    Article  Google Scholar 

  10. Ren, X. et al. Regulatory B cells limit CNS inflammation and neurologic deficits in murine experimental stroke. J. Neurosci. 31, 8556–8563 (2011).

    Article  CAS  Google Scholar 

  11. Shichita, T. et al. Pivotal role of cerebral interleukin-17-producing γδT cells in the delayed phase of ischemic brain injury. Nat. Med. 15, 946–950 (2009).

    Article  CAS  Google Scholar 

  12. Konoeda, F. et al. Therapeutic effect of IL-12/23 and their signaling pathway blockade on brain ischemia model. Biochem. Biophys. Res. Commun. 402, 500–506 (2010).

    Article  CAS  Google Scholar 

  13. Tang, S.C. et al. Pivotal role for neuronal Toll-like receptors in ischemic brain injury and functional deficits. Proc. Natl. Acad. Sci. USA 104, 13798–13803 (2007).

    Article  CAS  Google Scholar 

  14. Chen, C.J. et al. Identification of a key pathway required for the sterile inflammatory response triggered by dying cells. Nat. Med. 13, 851–856 (2007).

    Article  CAS  Google Scholar 

  15. Yanai, H. et al. HMGB proteins function as universal sentinels for nucleic-acid-mediated innate immune responses. Nature 462, 99–103 (2009).

    Article  CAS  Google Scholar 

  16. Marsh, B.J., Williams-Karnesky, R.L. & Stenzel-Poore, M.P. Toll-like receptor signaling in endogenous neuroprotection and stroke. Neuroscience 158, 1007–1020 (2009).

    Article  CAS  Google Scholar 

  17. Stewart, C.R. et al. CD36 ligands promote sterile inflammation through assembly of a Toll-like receptor 4 and 6 heterodimer. Nat. Immunol. 11, 155–161 (2010).

    Article  CAS  Google Scholar 

  18. Zhang, Q. et al. Circulating mitochondrial DAMPs cause inflammatory responses to injury. Nature 464, 104–107 (2010).

    Article  CAS  Google Scholar 

  19. Rivest, S. Regulation of innate immune responses in the brain. Nat. Rev. Immunol. 9, 429–439 (2009).

    Article  CAS  Google Scholar 

  20. Zhang, J. et al. Anti–high mobility group box-1 monoclonal antibody protects the blood-brain barrier from ischemia-induced disruption in rats. Stroke 42, 1420–1428 (2011).

    Article  CAS  Google Scholar 

  21. Kim, J.B. et al. HMGB1, a novel cytokine-like mediator linking acute neuronal death and delayed neuroinflammation in the postischemic brain. J. Neurosci. 26, 6413–6421 (2006).

    Article  CAS  Google Scholar 

  22. Yang, Q.W. et al. HMBG1 mediates ischemia-reperfusion injury by TRIF-adaptor independent Toll-like receptor 4 signaling. J. Cereb. Blood Flow Metab. 31, 593–605 (2011).

    Article  CAS  Google Scholar 

  23. Hayakawa, K., Qiu, J. & Lo, E.H. Biphasic actions of HMGB1 signaling in inflammation and recovery after stroke. Ann. NY Acad. Sci. 1207, 50–57 (2010).

    Article  CAS  Google Scholar 

  24. Riddell, J.R., Wang, X.Y., Minderman, H. & Gollnick, S.O. Peroxiredoxin 1 stimulates secretion of proinflammatory cytokines by binding to TLR4. J. Immunol. 184, 1022–1030 (2010).

    Article  CAS  Google Scholar 

  25. Chesterman, E.S. et al. Investigation of Prx1 protein expression provides evidence for conservation of cardiac-specific posttranscriptional regulation in vertebrates. Dev. Dyn. 222, 459–470 (2001).

    Article  CAS  Google Scholar 

  26. Patenaude, A., Murthy, M.R. & Mirault, M.E. Emerging roles of thioredoxin cycle enzymes in the central nervous system. Cell. Mol. Life Sci. 62, 1063–1080 (2005).

    Article  CAS  Google Scholar 

  27. Hu, X. et al. Peroxiredoxin-2 protects against 6-hydroxydopamine–induced dopaminergic neurodegeneration via attenuation of the apoptosis signal-regulating kinase (ASK1) signaling cascade. J. Neurosci. 31, 247–261 (2011).

    Article  CAS  Google Scholar 

  28. Rashidian, J. et al. Essential role of cytoplasmic cdk5 and Prx2 in multiple ischemic injury models, in vivo. J. Neurosci. 29, 12497–12505 (2009).

    Article  CAS  Google Scholar 

  29. Wood, Z.A., Schröder, E., Robin Harris, J. & Poole, L.B. Structure, mechanism and regulation of peroxiredoxins. Trends Biochem. Sci. 28, 32–40 (2003).

    Article  CAS  Google Scholar 

  30. Seo, M.S. et al. Identification of a new type of mammalian peroxiredoxin that forms an intramolecular disulfide as a reaction intermediate. J. Biol. Chem. 275, 20346–20354 (2000).

    Article  CAS  Google Scholar 

  31. Toohey, J.I. Sulfhydryl dependence in primary explant hematopoietic cells. Inhibition of growth in vitro with vitamin B12 compounds. Proc. Natl. Acad. Sci. USA 72, 73–77 (1975).

    Article  CAS  Google Scholar 

  32. Chou, J.L. et al. Proteomic investigation of a neural substrate intimately related to brain death. Proteomics 11, 239–248 (2011).

    Article  CAS  Google Scholar 

  33. Dayon, L. et al. Brain extracellular fluid protein changes in acute stroke patients. J. Proteome Res. 10, 1043–1051 (2011).

    Article  CAS  Google Scholar 

  34. Jin, M.H. et al. Characterization of neural cell types expressing peroxiredoxins in mouse brain. Neurosci. Lett. 381, 252–257 (2005).

    Article  CAS  Google Scholar 

  35. Brea, D. et al. Toll-like receptors 2 and 4 in ischemic stroke: outcome and therapeutic values. J. Cereb. Blood Flow Metab. 31, 1424–1431 (2011).

    Article  CAS  Google Scholar 

  36. Hirotsu, S. et al. Crystal structure of a multifunctional 2-Cys peroxiredoxin heme-binding protein 23kDa/proliferation-associated gene product. Proc. Natl. Acad. Sci. USA 96, 12333–12338 (1999).

    Article  CAS  Google Scholar 

  37. Declercq, J.P. et al. Crystal structure of human peroxiredoxin 5, a novel type of mammalian peroxiredoxin at 1.5 A resolution. J. Mol. Biol. 311, 751–759 (2001).

    Article  CAS  Google Scholar 

  38. Choi, H.J. et al. Crystal structure of a novel human peroxidase enzyme at 2.0 A resolution. Nat. Struct. Biol. 5, 400–406 (1998).

    Article  CAS  Google Scholar 

  39. Chen, G.Y. & Nuñez, G. Sterile inflammation: sensing and reacting to damage. Nat. Rev. Immunol. 10, 826–837 (2010).

    Article  CAS  Google Scholar 

  40. Yu, M. et al. HMGB1 signals through Toll-like receptor (TLR) 4 and TLR2. Shock 26, 174–179 (2006).

    Article  CAS  Google Scholar 

  41. Triantafilou, M. et al. Membrane sorting of Toll-like receptor (TLR)-2/6 and TLR2/1 heterodimers at the cell surface determines heterotypic associations with CD36 and intracellular targeting. J. Biol. Chem. 281, 31002–31011 (2006).

    Article  CAS  Google Scholar 

  42. Yi, H. et al. Pattern recognition scavenger receptor SRA/CD204 down-regulates Toll-like receptor 4 signaling–dependent CD8 T-cell activation. Blood 113, 5819–5828 (2009).

    Article  CAS  Google Scholar 

  43. Akashi-Takamura, S. & Miyake, K. TLR accessory molecules. Curr. Opin. Immunol. 20, 420–425 (2008).

    Article  CAS  Google Scholar 

  44. Eismann, T. et al. Peroxiredoxin-6 protects against mitochondrial dysfunction and liver injury during ischemia-reperfusion in mice. Am. J. Physiol. Gastrointest. Liver Physiol. 296, G266–G274 (2009).

    Article  CAS  Google Scholar 

  45. Liu, K. et al. Anti-high mobility group box 1 monoclonal antibody ameliorates brain infarction induced by transient ischemia in rats. FASEB J. 21, 3904–3916 (2007).

    Article  CAS  Google Scholar 

  46. Qiu, J. et al. Early release of HMGB-1 from neurons after the onset of brain ischemia. J. Cereb. Blood Flow Metab. 28, 927–938 (2008).

    Article  CAS  Google Scholar 

  47. Sugimori, H. et al. Krypton laser-induced photothrombotic distal middle cerebral artery occlusion without craniectomy in mice. Brain Res Brain Res. Protoc. 13, 189–196 (2004).

    Article  Google Scholar 

Download references

Acknowledgements

We thank N. Saito, N. Shiino and M. Asakawa for technical assistance. This work was supported by special grants-in-aid (A) and open research for young academics and specialists from the Ministry of Education, Culture, Sports, Science and Technology of Japan, PRESTO and CREST from the Japan Science and Technology Agency, an Intramural Research Grant (22-4) for Neurological and Psychiatric Disorders of the National Center of Neurology and Psychiatry (NCNP), the SENSHIN Research Foundation, the Takeda Science Foundation, the Uehara Memorial Foundation, the Mochida Memorial Foundation, the Scientific Research Fund from the Ministry of Health, Labor and Welfare of Japan (09156274) and the Program for Promotion of Fundamental Studies in Health Science of the National Institute of Biomedical Innovation.

Author information

Authors and Affiliations

Authors

Contributions

T. Shichita designed and performed experiments, analyzed data and wrote the manuscript; E.H. and A.K. performed TLR-deficient mouse analysis; R.M., R.S. and T. Sekiya participated in data analysis and discussion; I.T. provided specific input on protein analysis; H.O. and T.K. provided technical advice about experimental design; T.Y. and T.I. provided crucial input on Prx1's functions; H.T., S.M. and M.N. provided the HMGB1-specific antibody and crucial input on HMGB1; K.K. provided specific input regarding LC-MS analysis; K.M. and S.A. provided TLR2 and/or TLR4-deficient mice; A.Y. initiated and directed the entire study, designed experiments and wrote the manuscript.

Corresponding author

Correspondence to Akihiko Yoshimura.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–19, Supplementary Tables 1–5 and Supplementary Methods (PDF 1409 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Shichita, T., Hasegawa, E., Kimura, A. et al. Peroxiredoxin family proteins are key initiators of post-ischemic inflammation in the brain. Nat Med 18, 911–917 (2012). https://doi.org/10.1038/nm.2749

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nm.2749

This article is cited by

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing