Abstract

Type 2 immunity is critical for defense against cutaneous infections but also underlies the development of allergic skin diseases. We report the identification in normal mouse dermis of an abundant, phenotypically unique group 2 innate lymphoid cell (ILC2) subset that depended on interleukin 7 (IL-7) and constitutively produced IL-13. Intravital multiphoton microscopy showed that dermal ILC2 cells specifically interacted with mast cells, whose function was suppressed by IL-13. Treatment of mice deficient in recombination-activating gene 1 (Rag1−/−) with IL-2 resulted in the population expansion of activated, IL-5-producing dermal ILC2 cells, which led to spontaneous dermatitis characterized by eosinophil infiltrates and activated mast cells. Our data show that ILC2 cells have both pro- and anti-inflammatory properties and identify a previously unknown interactive pathway between two innate populations of cells of the immune system linked to type 2 immunity and allergic diseases.

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Acknowledgements

We thank J. Ho Cho and J. Sprent (Garvan Institute) for mice deficient in IL-7, IL-15 or Jak3; M. Kleinschek (DNAX) for Il25−/− mice; P. Besmer (Sloan Kettering Institute) for c-Kit–eGFP mice; Z. Eshar (Weizmann Institute of Science) for mouse SPE-7 hybridoma cells that produce IgE monoclonal antibody specific for 2,4-dinitrophenyl; A. Smith, S. Allen, S. Dervish, C. Zhu, A. Terry, Y. Wen Loh, K. Price and M. Camberis for technical assistance; M. Rizk and J. Qin for animal husbandry; L. Feigenbaum for help in preparing mice with transgenic expression of a bacterial artificial chromosome; N. Kolesnikoff and H. Taing for help with culturing bone marrow–derived mast cells; and L. Cavanagh for administrative assistance. Supported by the Australian National Health and Medical Research Council (M.A.G.), the Health Research Council of New Zealand, the Marjorie Barclay Trust, the Division of Intramural Research of the National Institute of Allergy and Infectious Diseases (US National Institutes of Health) and the Cancer Institute New South Wales (W.W.).

Author information

Author notes

    • Nital Sumaria

    Present address: Centre for Immunology and Infectious Disease, Blizard Institute, Barts and The London School of Medicine and Dentistry, London, UK.

Affiliations

  1. The Centenary Institute, Newtown, Australia.

    • Ben Roediger
    • , Nital Sumaria
    • , Thomas V Guy
    • , Andrew J Mitchell
    • , Szun S Tay
    • , Rohit Jain
    • , Philip L Tong
    • , Holly A Bolton
    • , Barbara Fazekas de St Groth
    •  & Wolfgang Weninger
  2. Malaghan Institute of Medical Research, Wellington, New Zealand.

    • Ryan Kyle
    • , Elizabeth Forbes-Blom
    •  & Graham Le Gros
  3. Centre for Cancer Biology, SA Pathology, Adelaide, South Australia, Australia.

    • Kwok Ho Yip
    •  & Michele A Grimbaldeston
  4. Department of Microbiology, Perelman School of Medicine Philadelphia, Pennsylvania, USA.

    • Brian S Kim
    •  & David Artis
  5. Institute for Immunology, Perelman School of Medicine Philadelphia, Pennsylvania, USA.

    • Brian S Kim
    •  & David Artis
  6. Department of Dermatology, Perelman School of Medicine Philadelphia, Pennsylvania, USA.

    • Brian S Kim
  7. National Institute of Allergic Disease, National Institutes of Health, Bethesda, Maryland, USA.

    • Xi Chen
    •  & William E Paul
  8. Discipline of Dermatology, University of Sydney, Camperdown, Australia.

    • Philip L Tong
    • , Barbara Fazekas de St Groth
    •  & Wolfgang Weninger
  9. Department of Dermatology, Royal Prince Alfred Hospital, Camperdown, Australia.

    • Philip L Tong
    •  & Wolfgang Weninger
  10. Department of Pathobiology, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, USA.

    • David Artis

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Contributions

B.R., W.W. and G.LG. conceived of the idea for this project and wrote the paper; B.R., R.K. and N.S. did immunology and flow cytometry experiments; M.A.G. provided expertise in mast-cell biology and, together with K.H.Y., did mast cell–stimulation experiments; B.R., R.J. and P.L.T. designed and did imaging experiments; E.F.-B., A.J.M., S.S.T., T.V.G., H.A.B., B.S.K., D.A. and B.F.d.S.G. contributed to experimental design and did experiments; X.C. and W.E.P. generated the 4C13R mice; and all authors discussed the results and commented on the paper.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to Graham Le Gros or Wolfgang Weninger.

Supplementary information

PDF files

  1. 1.

    Supplementary Text and Figures

    Supplementary Figures 1–8

Videos

  1. 1.

    Supplementary Video 1

    Representative video of mixed chimeric mouse skin (described in Fig. 4d) by intravital multiphoton microscopy. eGFP+ dILC2 shown in green. Blood vessels were visualized using Evans blue (red). Extracellular matrix in the dermis was detected by second harmonic generation (SHG) signals (blue). Video represents a z-projection through a volume of 28 μm within the dermis. Time shown in mm:ss. Data are representative of 2 independent experiments (n = 5).

  2. 2.

    Supplementary Video 2

    Representative video of mixed chimeric mouse skin (described in Fig. 4d) by intravital multiphoton microscopy. eGFP+ dILC2 shown in green. SHG signals shown in blue. Video represents a z-projection through a volume of 28 μm within the dermis. Time shown in mm:ss. Data are representative of 2 independent experiments (n = 5).

  3. 3.

    Supplementary Video 3

    Representative video of mixed chimeric mouse skin (described in Fig. 4d) by intravital multiphoton microscopy. eGFP+ dILC2 shown in green. mTomato+ cells shown in red. Blood vessels were visualized using Evans blue (white). SHG signals shown in blue. Video represents a z-projection through a volume of 28 μm within the dermis. Time shown in mm:ss. Data are representative of 2 independent experiments (n = 5).

  4. 4.

    Supplementary Video 4

    Representative video of mixed chimeric mouse skin (described in Fig. 4d) by intravital multiphoton microscopy. eGFP+ dILC2 shown in green. mTomato+ cells shown in red. SHG signals shown in blue. Video represents a z-projection through a volume of 28 μm within the dermis. Time shown in mm:ss. Data are representative of 2 independent experiments (n = 5).

  5. 5.

    Supplementary Video 5

    Representative video of albino C57BL/6 mice 8 weeks after irradiation and co-transfer of bone marrow from Rag1−/− Cxcr6+/gfp, mG/mT and CD11c-eYFP mice, as imaged by intravital multiphoton microscopy. eGFP+ dILC2 shown in green. eYFP+ dermal dendritic cell show in yellow. mTomato+ cells shown in red. Blood vessels were visualized using Evans blue (white). SHG signals shown in blue. Video represents a z-projection through a volume of 28 μm within the dermis. Time shown in mm:ss. Data are representative of 2 independent experiments (n = 4).

  6. 6.

    Supplementary Video 6

    Representative video of Brainbow32 mouse skin by intravital multiphoton microscopy. RFP fluorescence has been pseudocoloured such that RFPhi cells appear yellow while RFPdim cells are red. Blood vessels were visualized using Evans blue (white). Right: Close up of migratory RFPdim cells. Video represents a z-projection through a volume of 28 μm within the dermis. Time shown in mm:ss. Data are representative of 3 independent experiments (n = 3).

  7. 7.

    Supplementary Video 7

    Representative video of BrainbowAA Cxcr6+/gfp mouse skin by intravital multiphoton microscopy. RFPhi mast cells shown in red. eGFP+ cells shown in green. Video represents a z-projection through a volume of 28 μm within the dermis. Time shown in mm:ss. Data are representative of 2 independent experiments (n = 2).

  8. 8.

    Supplementary Video 8

    [Representative video of mixed chimeric mouse skin (described in Fig. 5e) by intravital multiphoton microscopy. eGFP+ dILC2 shown in green. RFPhi mast cells shown in red. Video represents a z-projection through a volume of 28 μm within the dermis. Time shown in mm:ss. Data are representative of 2 independent experiments (n = 4).

  9. 9.

    Supplementary Video 9

    Representative video of mixed chimeric mouse skin (described in Fig. 5e) by intravital multiphoton microscopy. eGFP+ dILC2 shown in green. RFPhi mast cells shown in red. Video represents a z-projection through a volume of 28 μm within the dermis. Time shown in mm:ss. Data are representative of 2 independent experiments (n = 4).

  10. 10.

    Supplementary Video 10

    Representative video of mixed chimeric mouse skin (described in Fig. 5e) by intravital multiphoton microscopy. eGFP+ dILC2 shown in green. RFPhi mast cells shown in red. Boxes indicate dILC2 that remained in close proximity with mast cells throughout the observation period. Video represents a z-projection through a volume of 28 μm within the dermis. Time shown in mm:ss. Data are representative of 2 independent experiments (n = 4).

  11. 11.

    Supplementary Video 11

    Representative video of IL-2-treated Rag1−/− Cxcr6+/gfp mice (described in Fig. 7b) by intravital multiphoton microscopy. eGFP+ dILC2 shown in green. Blood vessels were visualized using Evans blue (red). Extracellular matrix in the dermis was detected by SHG signals (blue). Video represents a z-projection through a volume of 28 μm within the dermis. Time shown in mm:ss. Data are representative of 2 independent experiments (n = 2).

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DOI

https://doi.org/10.1038/ni.2584

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