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IL-1β, IL-4 and IL-12 control the fate of group 2 innate lymphoid cells in human airway inflammation in the lungs

Nature Immunology volume 17, pages 636–645 (2016)Cite this article

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Abstract

Group 2 innate lymphoid cells (ILC2s) secrete type 2 cytokines, which protect against parasites but can also contribute to a variety of inflammatory airway diseases. We report here that interleukin 1β (IL-1β) directly activated human ILC2s and that IL-12 induced the conversion of these activated ILC2s into interferon-γ (IFN-γ)-producing ILC1s, which was reversed by IL-4. The plasticity of ILCs was manifested in diseased tissues of patients with severe chronic obstructive pulmonary disease (COPD) or chronic rhinosinusitis with nasal polyps (CRSwNP), which displayed IL-12 or IL-4 signatures and the accumulation of ILC1s or ILC2s, respectively. Eosinophils were a major cellular source of IL-4, which revealed cross-talk between IL-5-producing ILC2s and IL-4-producing eosinophils. We propose that IL-12 and IL-4 govern ILC2 functional identity and that their imbalance results in the perpetuation of type 1 or type 2 inflammation.

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Figure 1: Phenotype and tissue distribution of human ILCs, and changes in ILC ratios in COPD and nasal polyps.
Figure 2: Transferred ILC2s alter their phenotype and transcription-factor profile in Il2rg−/− NOD-SCID mice engrafted with human immune cells.
Figure 3: IL-1β activates ILC2s.
Figure 4: IL-12 governs the transdifferentiation of ILC2s into IFN-γ-producing cells.
Figure 5: ILC2 clones differentiate into IFN-γ-producing ILC1s.
Figure 6: IL-4 is essential for maintenance of CRTH2 expression and expansion of ILC2 populations.
Figure 7: Anti-IL-12p70 treatment abrogates the conversion of ILC2s into ILC1-like cells.
Figure 8: Eosinophils and ILC2s activate each other and perpetuate type 2 inflammation in nasal polyps from patients with CRS.

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References

  1. Barnes, P.J. Immunology of asthma and chronic obstructive pulmonary disease. Nat. Rev. Immunol. 8, 183–192 (2008).

    CAS  PubMed  Google Scholar 

  2. Hulse, K.E., Stevens, W.W., Tan, B.K. & Schleimer, R.P. Pathogenesis of nasal polyposis. Clin. Exp. Allergy 45, 328–346 (2015).

    CAS  PubMed  PubMed Central  Google Scholar 

  3. Nagarkar, D.R. et al. Thymic stromal lymphopoietin activity is increased in nasal polyps of patients with chronic rhinosinusitis. J. Allergy. Clin. Immunol. 132, 593–600 (2013).

    CAS  PubMed  PubMed Central  Google Scholar 

  4. Reh, D.D., Wang, Y., Ramanathan, M. Jr. & Lane, A.P. Treatment-recalcitrant chronic rhinosinusitis with polyps is associated with altered epithelial cell expression of interleukin-33. Am. J. Rhinol. Allergy 24, 105–109 (2010).

    PubMed  Google Scholar 

  5. Barnes, P.J. The cytokine network in chronic obstructive pulmonary disease. Am. J. Respir. Cell Mol. Biol. 41, 631–638 (2009).

    CAS  PubMed  Google Scholar 

  6. Kearley, J. et al. Cigarette smoke silences innate lymphoid cell function and facilitates an exacerbated type I interleukin-33-dependent response to infection. Immunity 42, 566–579 (2015).

    CAS  PubMed  Google Scholar 

  7. Holtzman, M.J., Byers, D.E., Alexander-Brett, J. & Wang, X. The role of airway epithelial cells and innate immune cells in chronic respiratory disease. Nat. Rev. Immunol. 14, 686–698 (2014).

    CAS  PubMed  PubMed Central  Google Scholar 

  8. McKenzie, A.N., Spits, H. & Eberl, G. Innate lymphoid cells in inflammation and immunity. Immunity 41, 366–374 (2014).

    CAS  PubMed  Google Scholar 

  9. Artis, D. & Spits, H. The biology of innate lymphoid cells. Nature 517, 293–301 (2015).

    CAS  PubMed  Google Scholar 

  10. Buonocore, S. et al. Innate lymphoid cells drive interleukin-23-dependent innate intestinal pathology. Nature 464, 1371–1375 (2010).

    CAS  PubMed  PubMed Central  Google Scholar 

  11. Klose, C.S. et al. A T-bet gradient controls the fate and function of CCR6-RORγt+ innate lymphoid cells. Nature 494, 261–265 (2013).

    CAS  PubMed  Google Scholar 

  12. Bernink, J.H. et al. Human type 1 innate lymphoid cells accumulate in inflamed mucosal tissues. Nat. Immunol. 14, 221–229 (2013).

    CAS  PubMed  Google Scholar 

  13. Bartemes, K.R., Kephart, G.M., Fox, S.J. & Kita, H. Enhanced innate type 2 immune response in peripheral blood from patients with asthma. J Allergy Clin Immunol 134, 671–678 (2014).

    CAS  PubMed  PubMed Central  Google Scholar 

  14. Nagakumar, P. et al. Type 2 innate lymphoid cells in induced sputum from children with severe asthma. J. Allergy Clin. Immunol. 137, 624–626.e6 (2015).

    PubMed  Google Scholar 

  15. 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).

    PubMed  Google Scholar 

  16. 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).

    CAS  PubMed  PubMed Central  Google Scholar 

  17. Teunissen, M.B. et al. Composition of innate lymphoid cell subsets in the human skin: enrichment of NCR+ ILC3 in lesional skin and blood of psoriasis patients. J. Invest. Dermatol. 134, 2351–2360 (2014).

    CAS  PubMed  Google Scholar 

  18. Kim, H.Y. et al. Interleukin-17-producing innate lymphoid cells and the NLRP3 inflammasome facilitate obesity-associated airway hyperreactivity. Nat. Med. 20, 54–61 (2014).

    CAS  PubMed  Google Scholar 

  19. Gasteiger, G., Fan, X., Dikiy, S., Lee, S.Y. & Rudensky, A.Y. Tissue residency of innate lymphoid cells in lymphoid and nonlymphoid organs. Science 350, 981–985 (2015).

    CAS  PubMed  PubMed Central  Google Scholar 

  20. Bernink, J.H. et al. Interleukin-12 and -23 control plasticity of CD127+ group 1 and group 3 innate lymphoid cells in the intestinal lamina propria. Immunity 43, 146–160 (2015).

    CAS  PubMed  Google Scholar 

  21. Vonarbourg, C. et al. Regulated expression of nuclear receptor RORγt confers distinct functional fates to NK cell receptor-expressing RORγt+ innate lymphocytes. Immunity 33, 736–751 (2010).

    CAS  PubMed  PubMed Central  Google Scholar 

  22. Hazenberg, M.D. & Spits, H. Human innate lymphoid cells. Blood 124, 700–709 (2014).

    CAS  PubMed  Google Scholar 

  23. Roediger, B. & Weninger, W. Group 2 innate lymphoid cells in the regulation of immune responses. Adv. Immunol. 125, 111–154 (2015).

    CAS  PubMed  Google Scholar 

  24. Brestoff, J.R. et al. Group 2 innate lymphoid cells promote beiging of white adipose tissue and limit obesity. Nature 519, 242–246 (2015).

    CAS  PubMed  Google Scholar 

  25. Mjösberg, J. et al. The transcription factor GATA3 is essential for the function of human type 2 innate lymphoid cells. Immunity 37, 649–659 (2012).

    PubMed  Google Scholar 

  26. Gimeno, R. et al. Monitoring the effect of gene silencing by RNA interference in human CD34+ cells injected into newborn RAG2−/− γc−/− mice: functional inactivation of p53 in developing T cells. Blood 104, 3886–3893 (2004).

    CAS  PubMed  Google Scholar 

  27. Hughes, T. et al. Interleukin-1β selectively expands and sustains interleukin-22+ immature human natural killer cells in secondary lymphoid tissue. Immunity 32, 803–814 (2010).

    CAS  PubMed  PubMed Central  Google Scholar 

  28. Cella, M. et al. A human natural killer cell subset provides an innate source of IL-22 for mucosal immunity. Nature 457, 722–725 (2009).

    CAS  PubMed  Google Scholar 

  29. Hackett, T.L., Shaheen, F., Zhou, S., Wright, J.L. & Churg, A. Fibroblast signal transducer and activator of transcription 4 drives cigarette smoke-induced airway fibrosis. Am. J. Respir. Cell Mol. Biol. 51, 830–839 (2014).

    PubMed  Google Scholar 

  30. Neurath, M.F., Finotto, S. & Glimcher, L.H. The role of Th1/Th2 polarization in mucosal immunity. Nat. Med. 8, 567–573 (2002).

    CAS  PubMed  Google Scholar 

  31. Miljkovic, D. et al. Association between group 2 innate lymphoid cells enrichment, nasal polyps and allergy in chronic rhinosinusitis. Allergy 69, 1154–1161 (2014).

    CAS  PubMed  Google Scholar 

  32. Voehringer, D., Shinkai, K. & Locksley, R.M. Type 2 immunity reflects orchestrated recruitment of cells committed to IL-4 production. Immunity 20, 267–277 (2004).

    CAS  PubMed  Google Scholar 

  33. Sojka, D.K. et al. Tissue-resident natural killer (NK) cells are cell lineages distinct from thymic and conventional splenic NK cells. eLife 3, e01659 (2014).

    PubMed  PubMed Central  Google Scholar 

  34. Silver, J.S. et al. Inflammatory triggers associated with COPD exacerbations orchestrate ILC2 plasticity in the lung. Nat. Immunol. doi:10.1038/ni.3443 (2016).10.1038/ni.3443

  35. Barnes, P.J. Therapeutic approaches to asthma-chronic obstructive pulmonary disease overlap syndromes. J. Allergy Clin. Immunol. 136, 531–545 (2015).

    PubMed  Google Scholar 

  36. Molofsky, A.B. et al. Interleukin-33 and interferon-γ counter-regulate group 2 innate lymphoid cell activation during immune perturbation. Immunity 43, 161–174 (2015).

    CAS  PubMed  PubMed Central  Google Scholar 

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

    CAS  PubMed  PubMed Central  Google Scholar 

  38. Southworth, T. et al. IFN-γ synergistically enhances LPS signalling in alveolar macrophages from COPD patients and controls by corticosteroid-resistant STAT1 activation. Br. J. Pharmacol. 166, 2070–2083 (2012).

    CAS  PubMed  PubMed Central  Google Scholar 

  39. Lee, B.J. et al. Protective effects of basic fibroblast growth factor in the development of emphysema induced by interferon-γ. Exp. Mol. Med. 43, 169–178 (2011).

    CAS  PubMed  PubMed Central  Google Scholar 

  40. Bachert, C. et al. Effect of subcutaneous dupilumab on nasal polyp burden in patients with chronic sinusitis and nasal polyposis: a randomized clinical trial. J. Am. Med. Assoc. 315, 469–479 (2016).

    CAS  Google Scholar 

  41. Wenzel, S. et al. Dupilumab in persistent asthma with elevated eosinophil levels. N. Engl. J. Med. 368, 2455–2466 (2013).

    CAS  PubMed  Google Scholar 

  42. Beck, L.A. et al. Dupilumab treatment in adults with moderate-to-severe atopic dermatitis. N. Engl. J. Med. 371, 130–139 (2014).

    PubMed  Google Scholar 

  43. Gevaert, P. et al. Nasal IL-5 levels determine the response to anti-IL-5 treatment in patients with nasal polyps. J. Allergy Clin. Immunol. 118, 1133–1141 (2006).

    CAS  PubMed  Google Scholar 

  44. Gevaert, P. et al. Mepolizumab, a humanized anti-IL-5 mAb, as a treatment option for severe nasal polyposis. J. Allergy Clin. Immunol. 128, 989–995 (2011).

    CAS  PubMed  Google Scholar 

  45. Corren, J. et al. Lebrikizumab treatment in adults with asthma. N. Engl. J. Med. 365, 1088–1098 (2011).

    CAS  PubMed  Google Scholar 

  46. Abt, M.C. et al. innate immune defenses mediated by two ILC subsets are critical for protection against acute Clostridium difficile infection. Cell Host Microbe 18, 27–37 (2015).

    CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

We thank B. Hooibrink for help with flow cytometry; staff of the Bloemenhove clinic in Heemstede, the Netherlands, for fetal tissues; K. Weijer for processing fetal material; B. Dierdorp and T. Dekker for performing the multiplex assay; E. van Rijnstra for help with the animal experiments; E. de Groot, D. van Egmond and Britt-Marie Nilsson for immunohistochemical staining; C. Loftus (Medetect AB) for computer simulations of spatial cell distributions. Y. Pineros for isolating eosinophils, and J. Fergusson for critical reading of the manuscript. Supported by the European Research Council (Advanced ERC grant 341038-AsthmaVir).

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Author notes

  1. Suzanne M Bal and Jochem H Bernink: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Cell Biology and Histology, Academic Medical Center, Amsterdam, the Netherlands

    Suzanne M Bal, Jochem H Bernink, Maho Nagasawa, Jelle Groot, Medya M Shikhagaie, Kornel Golebski, Melanie Bruchard, Hergen Spits & Xavier Romero Ros

  2. Department of Otorhinolaryngology, Academic Medical Center, Amsterdam, the Netherlands

    Kornel Golebski, Cornelis M van Drunen & Wytske Fokkens

  3. Department of Experimental Immunology, Academic Medical Center, Amsterdam, the Netherlands

    Rene Lutter

  4. Department of Pulmonology, Academic Medical Center, Amsterdam, the Netherlands

    Rene Lutter & Rene E Jonkers

  5. Sanquin Research and Landsteiner Laboratory, Amsterdam, the Netherlands

    Pleun Hombrink

  6. Department of Medical Microbiology, Academic Medical Center, Amsterdam, the Netherlands

    Julien Villaudy

  7. Department of Hematology, Academic Medical Center, Amsterdam, the Netherlands

    J Marius Munneke

  8. Department of Experimental Medical Sciences, Unit of Airway Inflammation, Lund University, Lund, Sweden

    Jonas S Erjefält

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Contributions

S.M.B. and J.H.B. designed the study, did experiments, analyzed the data and wrote the manuscript; M.N., J.G. and M.M.S. did experiments and analyzed the data; K.G., M.B., J.V. and J.M.M. did experiments; C.M.v.D. and W.F. provided nasal tissue; R.L. provided BAL fluid cells and analyzed data; R.E.J. and P.H. provided lung tissue; J.S.E. performed histological assessments; H.S. designed the study, analyzed data and wrote the manuscript ; X.R.R. designed the study, did experiments, analyzed the data and wrote the manuscript; and all authors critically read the manuscript.

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Correspondence to Hergen Spits.

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Integrated supplementary information

Supplementary Figure 1 Gating strategy for the detection of ILCs in human lungs and nasal polyps, and the expression of cell-surface proteins on nasal-polyp ILCs.

(a) Flow cytometry analysis of human lung and nasal polyp tissue of CD45, Lin (CD1a, CD3, CD14, CD16, CD19, CD34, CD123, BDCA2, FcɛR1α, TCRαβ, TCRγδ), CD127, CD161, c-Kit, CRTH2, and NKp44 to detect ILC1 (CRTH2- c-Kit- NKp44-), ILC2 (CRTH2+ c-Kit+/-), and ILC3 (CRTH2- c-Kit+ NKp44+/-). (b) Flow cytometry analyzing the expression of CRTH2, IL7Rα, CD161, ST2, TSLPR, IL17RB, CD25, KLRG1, IL4R, IL9R and ICOS in ILC1 (black), ILC2 (red) and ILC3 (grey line) from nasal polyp tissue. Isotype-matched control antibody is shown as grey shaded curve. Data are representative of at least three different donors (a,b).

Supplementary Figure 2 Repopulation of Il2rg–/– NOD-SCID mice with human hematopoietic stem cells and purity of expanded ILC2 populations.

(a) Flow cytometry analysis of human and murine CD45 expression in blood, lung and spleen of Il2rg–/– NOD-SCID mice eight weeks after engraftment with human hematopoietic stem cells. (b) c-Kit and CRTH2 expression after resorting of ILC2 that were expanded for four weeks with irradiated allogeneic peripheral blood mononuclear cells, irradiated Epstein-Barr virus–transformed JY human B cells, phytohemagglutinin and IL-2. (c) CRTH2 expression after culturing CTV labeled ILC2 for 4 days with IL-2. Data are representative of six mice (a) or five different experiments (b).

Supplementary Figure 3 Reactivation of ILC2 cells with IL-33 and TSLP and transdifferentiation of lung ILC2 cells.

(a) Flow cytometry analysis of c-Kit and CRTH2 expression on blood ILC2 (left panel) and IL-5 and IL-13 production (right panel) upon stimulation for five days with IL-2, IL-2 + IL-33 + TSLP. Cells were washed and stimulated for another four days with IL-2 + IL-33 + TSLP. (b) Expression of IL12RB2 on ILC1, ILC2, ILC3 and NK cells as measured by qPCR. (c) IL-5 and IL-13 production by blood ILC2, ILC1, and ILC2 expanded with IL-2, IL-1β and IL-12 after culture for 4 days with IL-2, IL-33, and TSLP, (d) intracellular IL-5 and IFN-γ production by cells as under cafter culture for 4 days with IL-2, IL-12, and IL-18. (e) Flow cytometry analysis of c-Kit and CRTH2 expression on lung ILC2 directly after isolation and upon stimulation with IL-2, IL-33, and IL-12 for seven days and IFN-γ production of lung c-Kit- ex-ILC2 obtained upon stimulation with IL-2, IL-33, and IL-12 for seven days. (f) Isolated ILC2 from lung cultured for 6 days either with IL-2, or IL-2, IL-33, and TSLP, or with IL-2, IL-33, TSLP, and IL-12. Cells were analyzed for c-Kit and CRTH2 expression (left panels), percentage of c-Kit- ex-ILC2 generated in above mentioned activation conditions (right panel). Data are representative of three (a,c,d,e,f) or four (b) different donors.

Supplementary Figure 4 Stimulation of ILC1 and ILC3 cells with IL-4, and gating strategy for BAL-fluid ILCs.

(a) Flow cytometry analysis of c-Kit and CRTH2 expression on blood ILC1 and ILC3 upon stimulation for five days with IL-2 or IL-2 and IL-4. (b) Gating strategy for ILC detection and ILC distribution in bronchoalveolar lavage (BAL) fluid. Data are representative of three individual experiments (a) or five different samples (b).

Supplementary Figure 5 Gating strategy for basophils, mast cells and eosinophils in nasal-polyp and turbinate tissue.

Polyp and turbinate tissue was analyzed by flow cytometry for CD45, CD3, CD123, FcɛRI, c-Kit, CD203c and Siglec8. Basophils were gated as CD3- CD123+ FcɛRI+ CD203c+, mast cells as c-Kit+ FcɛRI+ CD203c+, and eosinophils as SSChigh Siglec8+. Data are representative of five polyp and two turbinate samples.

Supplementary Figure 6 ILC2 spatial distribution.

Using the known x,y coordinates for each eosinophil, the true observed accumulated eosinophil counts within a fixed close space surrounding each ILC2 cell were calculated and compared to computer simulations of random ILC distributions. The true accumulated eosinophils count within a 300 µm radius around each of the 20 ILC in Figure 8b (n=668 ILC-associated eosinophils) was compared to the combined eosinophil counts calculated for simulated 250 000 cases of computer-generated random distributions of the same number of fictive ILCs (mean combined eosinophil count = 139, range 35-452).

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Bal, S., Bernink, J., Nagasawa, M. et al. IL-1β, IL-4 and IL-12 control the fate of group 2 innate lymphoid cells in human airway inflammation in the lungs. Nat Immunol 17, 636–645 (2016). https://doi.org/10.1038/ni.3444

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