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.

Pivotal role of cerebral interleukin-17–producing γδT cells in the delayed phase of ischemic brain injury

Nature Medicine volume 15, pages 946–950 (2009)Cite this article

Abstract

Lymphocyte recruitment and activation have been implicated in the progression of cerebral ischemia-reperfusion (I/R) injury, but the roles of specific lymphocyte subpopulations and cytokines during stroke remain to be clarified. Here we demonstrate that the infiltration of T cells into the brain, as well as the cytokines interleukin-23 (IL-23) and IL-17, have pivotal roles in the evolution of brain infarction and accompanying neurological deficits. Blockade of T cell infiltration into the brain by the immunosuppressant FTY720 reduced I/R-induced brain damage. The expression of IL-23, which was derived mostly from infiltrated macrophages, increased on day 1 after I/R, whereas IL-17 levels were elevated after day 3, and this induction of IL-17 was dependent on IL-23. These data, together with analysis of mice genetically disrupted for IL-17 and IL-23, suggest that IL-23 functions in the immediate stage of I/R brain injury, whereas IL-17 has an important role in the delayed phase of I/R injury during which apoptotic neuronal death occurs in the penumbra. Intracellular cytokine staining revealed that γδT lymphocytes, but not CD4+ helper T cells, were a major source of IL-17. Moreover, depletion of γδT lymphocytes ameliorated the I/R injury. We propose that T lymphocytes, including γδT lymphocytes, could be a therapeutic target for mitigating the inflammatory events that amplify the initial damage in cerebral ischemia.

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

Learn more

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

Figure 1: Effect of inflammatory cytokines on ischemic brain damage.
Figure 2: Infiltration of macrophages and T cells into the ischemic brain and their roles in inflammation.
Figure 3: Major producer cells of IL-23 and IL-17 in the ischemic brain.
Figure 4: The effects of γδT lymphocyte depletion in ischemic brain tissue.

Similar content being viewed by others

References

  1. Sacco, R.L., Chong, J.Y., Prabhakaran, S. & Elkind, M.S. Experimental treatments for acute ischaemic stroke. Lancet 369, 331–341 (2007).

    Article  CAS  Google Scholar 

  2. Ooboshi, H. et al. Postischemic gene transfer of interleukin-10 protects against both focal and global brain ischemia. Circulation 111, 913–919 (2006).

    Article  Google Scholar 

  3. Schroeter, M., Jander, S., Witte, O.W. & Stoll, G. Local immune responses in the rat middle cerebral artery occlusion. J. Neuroimmunol. 55, 195–203 (1994).

    Article  CAS  Google Scholar 

  4. Jander, S., Karemer, M., Schroeter, M., Witte, O.W. & Stoll, G. Lymphocytic infiltration and expression of intercellular adhesion molecule-1 in photochemically induced ischemia of the rat cortex. J. Cereb. Blood Flow Metab. 15, 42–51 (1995).

    Article  CAS  Google Scholar 

  5. 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 

  6. Lambertsen, K.L. et al. A role for interferon-gamma in focal cerebral ischemia in mice. J. Neuropathol. Exp. Neurol. 63, 942–955 (2004).

    Article  CAS  Google Scholar 

  7. Lock, C. et al. Gene-microarray analysis of multiple sclerosis lesions yields new targets validated in autoimmune encephalomyelitis. Nat. Med. 8, 500–508 (2002).

    Article  CAS  Google Scholar 

  8. Sutton, C., Brereton, C., Keogh, B., Mills, K.H. & Lavelle, E.C. A crucial role for interleukin (IL)-1 in the induction of IL-17-producing T cells that mediate autoimmune encephalomyelitis. J. Exp. Med. 203, 1685–1691 (2006).

    Article  CAS  Google Scholar 

  9. Park, H. et al. A distinct lineage of CD4 T cells regulates tissue inflammation by producing interleukin 17. Nat. Immunol. 6, 1133–1141 (2005).

    Article  CAS  Google Scholar 

  10. Langrish, C.L. et al. IL-23 drives a pathogenic T cell population that induces autoimmune inflammation. J. Exp. Med. 201, 233–240 (2005).

    Article  CAS  Google Scholar 

  11. Cua, D.J. et al. Interleukin-23 rather than interleukin-12 is the critical cytokine for autoimmune inflammation of the brain. Nature 421, 744–748 (2003).

    Article  CAS  Google Scholar 

  12. Chen, Y. et al. Anti-IL-23 therapy inhibits multiple inflammatory pathways and ameliorates autoimmune encephalomyelitis. J. Clin. Invest. 116, 1317–1326 (2006).

    Article  CAS  Google Scholar 

  13. Kebir, H. et al. Human TH17 lymphocytes promote blood-brain barrier disruption and central nervous system inflammation. Nat. Med. 13, 1173–1175 (2007).

    Article  CAS  Google Scholar 

  14. Ifergan, I. et al. The blood-brain barrier induces differentiation of migrating monocytes into Th17-polarizing dendritic cells. Brain 131, 785–799 (2008).

    Article  Google Scholar 

  15. Li, G.Z. et al. Expression of interleukin-17 in ischemic brain tissue. Scand. J. Immunol. 62, 481–486 (2005).

    Article  CAS  Google Scholar 

  16. Charriaut-Marlangue, C. et al. Apoptosis and necrosis after reversible focal ischemia: an in situ DNA fragmentation analysis. J. Cereb. Blood Flow Metab. 16, 186–194 (1996).

    Article  CAS  Google Scholar 

  17. Matsui, T. et al. Astrocytic activation and delayed infarct expansion after permanent focal ischemia in rats. Part I: enhanced astrocytic synthesis of s-100beta in the periinfarct area precedes delayed infarct expansion. J. Cereb. Blood Flow Metab. 22, 711–722 (2002).

    Article  CAS  Google Scholar 

  18. Campanella, M., Sciorati, C., Tarazzo, G. & Beltramo, M. Flow cytometric analysis of inflammatory cells in ischemic rat brain. Stroke 33, 586–592 (2002).

    Article  Google Scholar 

  19. Yamasaki, Y. et al. Interleukin-1 as a pathogenetic mediator of ischemic brain damage in rats. Stroke 26, 676–680 (1995).

    Article  CAS  Google Scholar 

  20. Romanic, A.M., White, R.F., Arleth, A.J., Ohlstein, E.H. & Barone, F.C. Matrix metalloproteinase expression increases after cerebral focal ischemia in rats: inhibition of matrix metalloproteinase-9 reduces infarct size. Stroke 29, 1020–1030 (1998).

    Article  CAS  Google Scholar 

  21. Yong, V.W., Power, C., Forsyth, P. & Edwards, D.R. Metalloproteinases in biology and pathology of the nervous system. Nat. Rev. Neurosci. 2, 502–511 (2001).

    Article  CAS  Google Scholar 

  22. Fujimoto, M. et al. Tissue inhibitor of metalloproteinases protect blood-brain barrier disruption in focal cerebral ischemia. J. Cereb. Blood Flow Metab. 28, 1674–1685 (2008).

    Article  CAS  Google Scholar 

  23. Friedlander, R.M. et al. Expression of a dominant negative mutant of interleukin-1 beta converting enzyme in transgenic mice prevents neuronal cell death induced by trophic factor withdrawal and ischemic brain injury. J. Exp. Med. 185, 933–940 (1997).

    Article  CAS  Google Scholar 

  24. Shibata, K., Yamada, H., Hara, H., Kishihara, K. & Yoshikai, Y. Resident Vδ1+γδ T cells control early infiltration of neutrophils after Escherichia coli infection via IL-17 production. J. Immunol. 178, 4466–4472 (2007).

    Article  CAS  Google Scholar 

  25. Jensen, K.D. et al. Thymic selection determines gammadelta T cell effector fate: antigen-naïve cells make interleukin-17 and antigen-experienced cells make interferon gamma. Immunity 29, 90–100 (2008).

    Article  CAS  Google Scholar 

  26. Nakamura, R. et al. Tyk2-signaling plays an important role in host defense against Escherichia coli through IL-23-induced IL-17 production by gammadelta T cells. J. Immunol. 181, 2071–2075 (2008).

    Article  CAS  Google Scholar 

  27. Gaudinski, M.R. et al. Establishing final infarct volume: stroke lesion evolution past 30 days is insignificant. Stroke 39, 2765–2768 (2008).

    Article  Google Scholar 

  28. Shimosegawa, E. et al. Metabolic penumbra of acute brain infarction: a correlation with infarct growth. Ann. Neurol. 57, 495–504 (2005).

    Article  Google Scholar 

  29. Liesz, A. et al. Regulatory T cells are key cerebroprotective immunomodulators in acute experimental stroke. Nat. Med. 15, 192–199 (2009).

    Article  CAS  Google Scholar 

  30. Nakae, S. et al. Antigen-specific T cell sensitization is impaired in IL-17-deficient mice, causing suppression of allergic cellular and humoral responses. Immunity 17, 375–387 (2002).

    Article  CAS  Google Scholar 

  31. Tanaka, K. et al. Loss of suppressor of cytokine signaling 1 in helper T cells leads to defective Th17 differentiation by enhancing antagonistic effects of IFN-gamma on STAT3 and Smads. J. Immunol. 180, 3746–3756 (2008).

    Article  CAS  Google Scholar 

  32. Hara, H. et al. Inhibition of interleukin 1beta converting enzyme family proteases reduces ischemic and excitotoxic neuronal damage. Proc. Natl. Acad. Sci. USA 94, 2007–2012 (1997).

    Article  CAS  Google Scholar 

  33. 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 S. Hamano for TCRγδ-specific antibody hybridoma, T. Yoshioka, N. Shiino and M. Asakawa for technical assistance, E. Ishikawa and H. Noguchi for technical support for human ischemic brain section, T. Kobayashi for discussion and technical help and N. Soma for manuscript preparation. This study was supported by special Grants-in-Aid from the Ministry of Education, Culture, Sports, Science, and Technology of Japan, the Program for Promotion of Fundamental Studies in Health Sciences of the National Institute of Biomedical Innovation (NIBIO, 07-4), the Naito Foundation and the Astellas Foundation for Research on Metabolic Disorders.

Author information

Authors and Affiliations

  1. Department of Microbiology and Immunology, School of Medicine, Keio University, Shinjuku-ku, Tokyo, Japan

    Takashi Shichita, Yuki Sugiyama, Ryusuke Nakagawa, Ichiro Takada & Akihiko Yoshimura

  2. Division of Molecular and Cellular Immunology, Medical Institute of Bioregulation, Kyushu University, Fukuoka, Japan

    Takashi Shichita

  3. Department of Medicine and Clinical Science, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan

    Takashi Shichita, Hiroaki Ooboshi, Hiroshi Sugimori & Mitsuo Iida

  4. Department of Internal Medicine, Fukuoka Dental College Medical and Dental Hospital, Fukuoka, Japan

    Hiroaki Ooboshi

  5. Department of Neuropathology, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan

    Toru Iwaki

  6. Department of Pathology, School of Medicine, Keio University, Shijuku-ku, Tokyo, Japan

    Yasunori Okada

  7. Department of Immunology, Schering-Plough, Palo Alto, California, USA

    Daniel J Cua

  8. Center for Experimental Medicine and Department of Cancer Biology, Institute of Medical Science, University of Tokyo, Tokyo, Japan

    Yoichiro Iwakura

  9. Japan Science and Technology Agency, Core Research for Evolutional Science and Technology (CREST), Tokyo, Japan

    Akihiko Yoshimura

Authors
  1. Takashi Shichita
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yuki Sugiyama
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Hiroaki Ooboshi
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Hiroshi Sugimori
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Ryusuke Nakagawa
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Ichiro Takada
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Toru Iwaki
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Yasunori Okada
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Mitsuo Iida
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Daniel J Cua
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Yoichiro Iwakura
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Akihiko Yoshimura
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

T.S. designed and performed experiments, analyzed data and wrote the manuscript; Y.S. performed experiments and analyzed data; H.O. contributed to the experimental design, analysis and manuscript writing; H.S. performed experiments; R.N. provided specific input into flow cytometry and mouse analysis, I.T. contributed to manuscript writing; T.I. and Y.O. contributed to histochemistry; M.I. contributed to the experimental design; D.J.C. and Y.I. provided mice and critical input on IL-23 and IL-17 functions; A.Y. initiated and directed the entire study, designed experiments and wrote the manuscript.

Corresponding author

Correspondence to Akihiko Yoshimura.

Supplementary information

Supplementary Text and Figures

Supplementary Figs. 1–7, Supplementary Tables 1–4 and Supplementary Methods (PDF 564 kb)

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Shichita, T., Sugiyama, Y., Ooboshi, H. 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). https://doi.org/10.1038/nm.1999

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nm.1999

This article is cited by

Search

Advanced 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