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:

IL-25-responsive, lineage-negative KLRG1hi cells are multipotential ‘inflammatory’ type 2 innate lymphoid cells

Nature Immunology volume 16, pages 161–169 (2015)Cite this article

Subjects

Abstract

Innate lymphoid cells (ILCs) are lymphocyte-like cells that lack T cell or B cell antigen receptors and mediate protective and repair functions through cytokine secretion. Among these, type 2 ILCs (ILC2 cells) are able to produce type 2 cytokines. We report the existence of an inflammatory ILC2 (iILC2) population responsive to interleukin 25 (IL-25) that complemented IL-33-responsive natural ILC2 (nILC2) cells. iILC2 cells developed into nILC2-like cells in vitro and in vivo and contributed to the expulsion of Nippostrongylus brasiliensis. They also acquired IL-17-producing ability and provided partial protection against Candida albicans. We propose that iILC2 cells are transient progenitors of ILCs mobilized by inflammation and infection that develop into nILC2-like cells or ILC3-like cells and contribute to immunity to both helminths and fungi.

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

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

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

Figure 1: IL-25 induces a Lin−ST2−KLRG1hi cell population distinct from nILC2 or MPPtype2 cells.
Figure 2: iILC2 development depends on γc and IL-7Rα.
Figure 3: iILC2 cells produce type 2 cytokines.
Figure 4: iILC2 cells develop into ST2+ nILC2-like cells both in vitro and in vivo.
Figure 5: iILC2 cells regulate anti-helminth immunity.
Figure 6: iILC2 cells express RORγt and have the ability to develop into IL-17-producing cells.
Figure 7: iILC2 cells contribute to antifungal immunity.

Similar content being viewed by others

References

  1. Walker, J.A., Barlow, J.L. & McKenzie, A.N. Innate lymphoid cells–how did we miss them? Nat. Rev. Immunol. 13, 75–87 (2013).

    Article  CAS  Google Scholar 

  2. Spits, H. & Cupedo, T. Innate lymphoid cells: emerging insights in development, lineage relationships, and function. Annu. Rev. Immunol. 30, 647–675 (2012).

    Article  CAS  Google Scholar 

  3. Spits, H. et al. Innate lymphoid cells–a proposal for uniform nomenclature. Nat. Rev. Immunol. 13, 145–149 (2013).

    CAS  Google Scholar 

  4. Moro, K. et al. Innate production of TH2 cytokines by adipose tissue-associated c-Kit+Sca-1+ lymphoid cells. Nature 463, 540–544 (2010).

    Article  CAS  Google Scholar 

  5. Neill, D.R. et al. Nuocytes represent a new innate effector leukocyte that mediates type-2 immunity. Nature 464, 1367–1370 (2010).

    Article  CAS  Google Scholar 

  6. Price, A.E. et al. Systemically dispersed innate IL-13-expressing cells in type 2 immunity. Proc. Natl. Acad. Sci. USA 107, 11489–11494 (2010).

    Article  CAS  Google Scholar 

  7. Saenz, S.A. et al. IL25 elicits a multipotent progenitor cell population that promotes TH2 cytokine responses. Nature 464, 1362–1366 (2010).

    Article  CAS  Google Scholar 

  8. Saenz, S.A. et al. IL-25 simultaneously elicits distinct populations of innate lymphoid cells and multipotent progenitor type 2 (MPPtype2) cells. J. Exp. Med. 210, 1823–1837 (2013).

    Article  CAS  Google Scholar 

  9. Fallon, P.G. et al. Identification of an interleukin (IL)-25-dependent cell population that provides IL-4, IL-5, and IL-13 at the onset of helminth expulsion. J. Exp. Med. 203, 1105–1116 (2006).

    Article  CAS  Google Scholar 

  10. Chang, Y.J. et al. Innate lymphoid cells mediate influenza-induced airway hyper-reactivity independently of adaptive immunity. Nat. Immunol. 12, 631–638 (2011).

    Article  CAS  Google Scholar 

  11. Motomura, Y. et al. Basophil-derived interleukin-4 controls the function of natural helper cells, a member of ILC2s, in lung inflammation. Immunity 40, 758–771 (2014).

    Article  CAS  Google Scholar 

  12. Salimi, M. et al. A role for IL-25 and IL-33-driven type-2 innate lymphoid cells in atopic dermatitis. J. Exp. Med. 210, 2939–2950 (2013).

    Article  CAS  Google Scholar 

  13. Monticelli, L.A. et al. Innate lymphoid cells promote lung-tissue homeostasis after infection with influenza virus. Nat. Immunol. 12, 1045–1054 (2011).

    Article  CAS  Google Scholar 

  14. McHedlidze, T. et al. Interleukin-33-dependent innate lymphoid cells mediate hepatic fibrosis. Immunity 39, 357–371 (2013).

    Article  CAS  Google Scholar 

  15. Nussbaum, J.C. et al. Type 2 innate lymphoid cells control eosinophil homeostasis. Nature 502, 245–248 (2013).

    Article  CAS  Google Scholar 

  16. Wilhelm, C. et al. An IL-9 fate reporter demonstrates the induction of an innate IL-9 response in lung inflammation. Nat. Immunol. 12, 1071–1077 (2011).

    Article  CAS  Google Scholar 

  17. Magri, G. et al. Innate lymphoid cells integrate stromal and immunological signals to enhance antibody production by splenic marginal zone B cells. Nat. Immunol. 15, 354–364 (2014).

    Article  CAS  Google Scholar 

  18. Boos, M.D., Yokota, Y., Eberl, G. & Kee, B.L. Mature natural killer cell and lymphoid tissue-inducing cell development requires Id2-mediated suppression of E protein activity. J. Exp. Med. 204, 1119–1130 (2007).

    Article  CAS  Google Scholar 

  19. Hoyler, T. et al. The transcription factor GATA-3 controls cell fate and maintenance of type 2 innate lymphoid cells. Immunity 37, 634–648 (2012).

    Article  CAS  Google Scholar 

  20. Yagi, R. et al. The transcription factor GATA3 is critical for the development of all IL-7Rα-expressing innate lymphoid cells. Immunity 40, 378–388 (2014).

    Article  CAS  Google Scholar 

  21. Wong, S.H. et al. Transcription factor RORα is critical for nuocyte development. Nat. Immunol. 13, 229–236 (2012).

    Article  CAS  Google Scholar 

  22. Yang, Q. et al. T cell factor 1 is required for group 2 innate lymphoid cell generation. Immunity 38, 694–704 (2013).

    Article  CAS  Google Scholar 

  23. Spooner, C.J. et al. Specification of type 2 innate lymphocytes by the transcriptional determinant Gfi1. Nat. Immunol. 14, 1229–1236 (2013).

    Article  CAS  Google Scholar 

  24. Hwang, Y.Y. & McKenzie, A.N. Innate lymphoid cells in immunity and disease. Adv. Exp. Med. Biol. 785, 9–26 (2013).

    Article  CAS  Google Scholar 

  25. Mjösberg, J.M. et al. Human IL-25- and IL-33-responsive type 2 innate lymphoid cells are defined by expression of CRTH2 and CD161. Nat. Immunol. 12, 1055–1062 (2011).

    Article  Google Scholar 

  26. Guo, L. et al. IL-1 family members and STAT activators induce cytokine production by Th2, Th17, and Th1 cells. Proc. Natl. Acad. Sci. USA 106, 13463–13468 (2009).

    Article  CAS  Google Scholar 

  27. Licona-Limón, P., Kim, L.K., Palm, N.W. & Flavell, R.A. TH2, allergy and group 2 innate lymphoid cells. Nat. Immunol. 14, 536–542 (2013).

    Article  Google Scholar 

  28. Gladiator, A., Wangler, N., Trautwein-Weidner, K. & LeibundGut-Landmann, S. Cutting edge: IL-17-secreting innate lymphoid cells are essential for host defense against fungal infection. J. Immunol. 190, 521–525 (2013).

    Article  CAS  Google Scholar 

  29. O'Shea, J.J. & Paul, W.E. Mechanisms underlying lineage commitment and plasticity of helper CD4+ T cells. Science 327, 1098–1102 (2010).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank J. Zhu for critical reading of the manuscript; J. Edwards for assistance in the preparation of sorter-purified cells; L. Feigenbaum of the SAIC Laboratory Animal Sciences Program for injection of the recombinant bacterial artificial chromosome into oocytes, transfer into pseudopregnant females and screening of pups; C. Dong (MD Anderson Cancer Center) for IL-17F–RFP reporter mice; A. McKenzie (MRC Laboratory of Molecular Biology, Cambridge) for Il1rl1−/− mice; and members of Laboratory of Immunology at NIAID for discussions. Supported by the NIAID Division of Intramural Research (US National Institutes of Health).

Author information

Authors and Affiliations

  1. Laboratory of Immunology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland, USA

    Yuefeng Huang, Liying Guo, Xi Chen, Jane Hu-Li & William E Paul

  2. Laboratory of Clinical Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland, USA

    Jin Qiu & Peter R Williamson

  3. Laboratory of Molecular Immunology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland, USA

    Ulrich Siebenlist

  4. Diet, Genomics, and Immunology Laboratory, Beltsville Human Nutrition Research Center, Agricultural Research Service, USDA, Beltsville, Maryland, USA

    Joseph F Urban Jr

Authors
  1. Yuefeng Huang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Liying Guo
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jin Qiu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Xi Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Jane Hu-Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Ulrich Siebenlist
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Peter R Williamson
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Joseph F Urban Jr
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. William E Paul
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

Y.H. and W.E.P. designed and interpreted the experiments and wrote the manuscript; L.G. assisted with the experiments and read the manuscript; Y.H. did the experiments; J.Q. did the C. albicans oral infection; X.C. generated 4C13R mice; J.H.-L. assisted with cell culture and flow cytometry; U.S. provided Il17rb−/− mice; P.R.W. assisted in the design and interpretation of experiments with C. albicans; and J.F.U. provided N. brasiliensis and helped to design and interpret experiments with N. brasiliensis.

Corresponding author

Correspondence to William E Paul.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Integrated supplementary information

Supplementary Figure 1 iILC2 cells are induced in several sites of mice by IL-25 treatment.

Wild-type B6 mice were treated i.p. with PBS or IL-25 daily for 3 days and leukocytes form various tissues were isolated and analyzed by flow cytometry for ST2, KLRG1 and lineage markers expression. Cells were gated from Lin–. Data are representative of two independent experiments (n=3 mice for each group in each experiment).

Supplementary Figure 2 A portion of nILC2 cells in naive mice produce IL-13.

(a) Map of recombined BAC transgene for 4C13R dual reporter mice. LCR, TH2 locus control region. (b) Lung leukocytes from wild-type B6 or 4C13R mice were isolated and analyzed by flow cytometry for expression of lineage, Thy1, ST2, KLRG1, AmCyan and DsRed. nILC2s were gated on Lin–Thy1hiST2+KLRG1int cells. Data are pooled from three independent experiments (n=6 wild-type mice, n=8 4C13R mice).

Source data

Supplementary Figure 3 iILC2–derived nILC2-like cells do not re-express IL-25 receptors upon IL-2 signaling.

iILC2s sorted from IL-25-treated Rag2–/– mice were cultured in IL-7 and IL-33 for 3 days, and then cells were stimulated with IL-2, or with IL-2 plus IL-25, for 2 days. IL-17RB expression was measured by flow cytometry. Data are representative of two independent experiments (n=3 cell culture wells for each condition in each experiment).

Supplementary Figure 4 iILC2 cells produce IL-13 and become nILC2-like cells during N. brasiliensis infection.

(a) 4C13R mice were infected with N. brasiliensis for various days and lung leukocytes were isolated and analyzed by flow cytometry for lineage, Thy1, KLRG1, AmCyan and DsRed expression. nILC2s were gated on Lin–Thy1hiKLRG1int (red) and iILC2s were gated on Lin–Thy1low KLRG1hi (blue). (b) 4 ×104 nILC2s sorted from naïve CD45.1 Rag1–/– mice or 20 ×104 iILC2s sorted from IL-25-treated CD45.1 Rag1–/– mice were transferred into Rag2–/–Il2rg–/– mice, which were infected with N. brasiliensis. 14 days later, transferred cells in the lung were analyzed by flow cytometry for expression of CD45.1, KLRG1, ST2 and Thy1. Data are representative of two independent experiments in a, b (n=3 mice for each group in each experiment of a, n=2 recipient mice in each experiment of b).

Supplementary Figure 5 Both IL-17-producing and non–IL-17-producing iILC2 cells can develop into IL-17–IL-13 double producers in vitro.

iILC2s, from IL-25-treated IL-17-RFP mice, were stimulated with PMA plus ionomycin and then were divided into RFP+ and RFP– populations through cell sorting. The purified populations were cultured in either “TH2” conditions or “TH17” conditions for 7 days. IL-13 and IL-17 production was measured by intracellular staining after 6-hour stimulation with PMA plus ionomycin. Data are representative of two independent experiments (n=3 cell culture wells for each condition in each experiment).

Supplementary Figure 6 nILC2 cells have less ability than iILC2 cells to develop into IL-17 producers.

nILC2s (sorted from naïve Rag2–/– mice) and iILC2s (sorted from IL-25-treated Rag2–/– mice) were cultured in various conditions as indicated. 7 days after culture, cells were stimulated with PMA plus ionomycin for 6 hours and the production of IL-13 and IL-17 were determined by intracellular cytokine staining. Data are representative of two independent experiments (n=2 cell culture wells for each condition in each experiment).

Supplementary Figure 7 C. albicans oral infection elicits ILCs in cervical lymph nodes.

Rag2–/– mice were infected sublingually with C. albicans for 5 days and leukocytes from cervical LNs were analyzed by flow cytometry for expression of lineage, Thy1, IL-7Rα, c-Kit, IL-17RB, ST2, and KLRG1. ILCs were gated as Lin–IL-7Rα+Thy1+. Data are representative of two independent experiments (n=3 mice for each group in each experiment).

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–7 (PDF 3530 kb)

Source data

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Huang, Y., Guo, L., Qiu, J. et al. IL-25-responsive, lineage-negative KLRG1hi cells are multipotential ‘inflammatory’ type 2 innate lymphoid cells. Nat Immunol 16, 161–169 (2015). https://doi.org/10.1038/ni.3078

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/ni.3078

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