Abstract

It remains largely unclear how antigen-presenting cells (APCs) encounter effector or memory T cells efficiently in the periphery. Here we used a mouse contact hypersensitivity (CHS) model to show that upon epicutaneous antigen challenge, dendritic cells (DCs) formed clusters with effector T cells in dermal perivascular areas to promote in situ proliferation and activation of skin T cells in a manner dependent on antigen and the integrin LFA-1. We found that DCs accumulated in perivascular areas and that DC clustering was abrogated by depletion of macrophages. Treatment with interleukin 1α (IL-1α) induced production of the chemokine CXCL2 by dermal macrophages, and DC clustering was suppressed by blockade of either the receptor for IL-1 (IL-1R) or the receptor for CXCL2 (CXCR2). Our findings suggest that the dermal leukocyte cluster is an essential structure for elicitating acquired cutaneous immunity.

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References

  1. 1.

    & Homing and cellular traffic in lymph nodes. Nat. Rev. Immunol. 3, 867–878 (2003).

  2. 2.

    et al. The vast majority of CLA+ T cells are resident in normal skin. J. Immunol. 176, 4431–4439 (2006).

  3. 3.

    et al. Langerin expressing cells promote skin immune responses under defined conditions. J. Immunol. 180, 4722–4727 (2008).

  4. 4.

    et al. Macrophages and neutrophils are the targets for immune suppression by glucocorticoids in contact allergy. J. Clin. Invest. 117, 1381–1390 (2007).

  5. 5.

    & The IL-1 family: regulators of immunity. Nat. Rev. Immunol. 10, 89–102 (2010).

  6. 6.

    , & Interleukin-1 and cutaneous inflammation: a crucial link between innate and acquired immunity. J. Invest. Dermatol. 114, 602–608 (2000).

  7. 7.

    et al. IL-1-induced tumor necrosis factor-α elicits inflammatory cell infiltration in the skin by inducing IFN-γ-inducible protein 10 in the elicitation phase of the contact hypersensitivity response. Int. Immunol. 15, 251–260 (2003).

  8. 8.

    , , , & Contact allergy to allergens of the TRUE-test (panels 1 and 2) has decreased modestly in the general population. Br. J. Dermatol. 161, 1124–1129 (2009).

  9. 9.

    et al. Migratory dermal dendritic cells act as rapid sensors of protozoan parasites. Plos Pathog 4, e1000222 (2008).

  10. 10.

    & Integrin inside-out signaling and the immunological synapse. Curr. Opin. Cell Biol. 24, 107–115 (2012).

  11. 11.

    et al. In vivo imaging of T-cell motility in the elicitation phase of contact hypersensitivity using two-photon microscopy. J. Invest. Dermatol. 131, 977–979 (2011).

  12. 12.

    et al. A new mutation, aly, that induces a generalized lack of lymph nodes accompanied by immunodeficiency in mice. Eur. J. Immunol. 24, 429–434 (1994).

  13. 13.

    et al. Role of mast cells and basophils in IgE responses and in allergic airway hyperresponsiveness. J. Immunol. 188, 1809–1818 (2012).

  14. 14.

    et al. Requirement of interaction between mast cells and skin dendritic cells to establish contact hypersensitivity. PLoS ONE 6, e25538 (2011).

  15. 15.

    & Early molecular events in the induction phase of contact sensitivity. Proc. Natl. Acad. Sci. USA 89, 1398–1402 (1992).

  16. 16.

    , , & Generation and characterization of murine alternatively activated macrophages. Methods Mol. Biol. 946, 225–239 (2013).

  17. 17.

    et al. CXCR2 blockade reduces radical formation in hyperoxia-exposed newborn rat lung. Pediatr. Res. 60, 299–303 (2006).

  18. 18.

    , , & Update of immune events in the murine contact hypersensitivity model: toward the understanding of allergic contact dermatitis. J. Invest. Dermatol. 133, 303–315 (2013).

  19. 19.

    , & Early immune events in the induction of allergic contact dermatitis. Nat. Rev. Immunol. 12, 114–124 (2012).

  20. 20.

    et al. Mechanical deformation promotes secretion of IL-1 alpha and IL-1 receptor antagonist. J. Immunol. 159, 5084–5088 (1997).

  21. 21.

    , , & Lymphocyte adhesion to psoriatic dermal endothelium is mediated by a tissue-specific receptor/ligand interaction. J. Invest. Dermatol. 91, 423–428 (1988).

  22. 22.

    , & IL-1 receptor signaling is required at multiple stages of sensitization and elicitation of the contact hypersensitivity response. J. Immunol. 188, 1761–1771 (2012).

  23. 23.

    et al. Interleukin-1 receptor antagonist suppresses contact hypersensitivity. J. Invest. Dermatol. 105, 334–338 (1995).

  24. 24.

    et al. The role of CXCR2 activity in the contact hypersensitivity response in mice. Eur. Cytokine Netw. 17, 42–48 (2006).

  25. 25.

    , , & Terminology: nomenclature of mucosa-associated lymphoid tissue. Mucosal Immunol. 1, 31–37 (2008).

  26. 26.

    Skin-associated lymphoid tissues (SALT): origins and functions. J. Invest. Dermatol. 80 (suppl.), 12s–16s (1983).

  27. 27.

    & Skin as a peripheral lymphoid organ: revisiting the concept of skin-associated lymphoid tissues. J. Invest. Dermatol. 131, 2178–2185 (2011).

  28. 28.

    et al. Role of inducible bronchus associated lymphoid tissue (iBALT) in respiratory immunity. Nat. Med. 10, 927–934 (2004).

  29. 29.

    et al. Dynamics and function of Langerhans cells in vivo: dermal dendritic cells colonize lymph node areas distinct from slower migrating Langerhans cells. Immunity 22, 643–654 (2005).

  30. 30.

    et al. In vivo depletion of CD11c+ dendritic cells abrogates priming of CD8+ T cells by exogenous cell-associated antigens. Immunity 17, 211–220 (2002).

  31. 31.

    et al. Visualizing dendritic cell networks in vivo. Nat. Immunol. 5, 1243–1250 (2004).

  32. 32.

    et al. Protective role of macrophages in noninflammatory lung injury caused by selective ablation of alveolar epithelial type II Cells. J. Immunol. 178, 5001–5009 (2007).

  33. 33.

    & Homeostasis of peripheral B cells in the absence of B cell influx from the bone marrow. J. Exp. Med. 194, 1151–1164 (2001).

  34. 34.

    et al. Production of mice deficient in genes for interleukin (IL)-1α, IL-1β, IL-1α/β, and IL-1 receptor antagonist shows that IL-1β is crucial in turpentine-induced fever development and glucocorticoid secretion. J. Exp. Med. 187, 1463–1475 (1998).

  35. 35.

    et al. Immunogenicity of whole-parasite vaccines against Plasmodium falciparum involves malarial hemozoin and host TLR9. Cell Host Microbe 7, 50–61 (2010).

  36. 36.

    , , , & Gout-associated uric acid crystals activate the NALP3 inflammasome. Nature 440, 237–241 (2006).

  37. 37.

    et al. Role of caspase-1 in experimental pneumococcal meningitis: evidence from pharmacologic caspase inhibition and caspase-1-deficient mice. Ann. Neurol. 51, 319–329 (2002).

  38. 38.

    et al. Activated regulatory T cells are the major T cell type emigrating from the skin during a cutaneous immune response in mice. J. Clin. Invest. 120, 883–893 (2010).

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Acknowledgements

We thank H. Yagita (Juntendo University) for the KBA neutralizing antibody to LFA-1; P. Bergstresser and J. Cyster for critical reading of our manuscript. Supported by grants-in-aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology of Japan.

Author information

Author notes

    • Yohei Natsuaki
    •  & Gyohei Egawa

    These authors contributed equally to this work.

Affiliations

  1. Department of Dermatology, Kyoto University Graduate School of Medicine, Kyoto, Japan.

    • Yohei Natsuaki
    • , Gyohei Egawa
    • , Satoshi Nakamizo
    • , Sachiko Ono
    • , Sho Hanakawa
    • , Nobuhiro Kusuba
    • , Atsushi Otsuka
    • , Akihiko Kitoh
    • , Tetsuya Honda
    • , Saeko Nakajima
    • , Yoshiki Miyachi
    •  & Kenji Kabashima
  2. Department of Dermatology, Kurume University School of Medicine, Fukuoka, Japan.

    • Yohei Natsuaki
    •  & Takashi Hashimoto
  3. Research Unit for Immunodynamics, RIKEN Research Center for Allergy and Immunology, Kanagawa, Japan.

    • Takaharu Okada
  4. Department of Pharmaceutical Biochemistry, Graduate School of Pharmaceutical Sciences, Kumamoto University, Kumamoto, Japan.

    • Soken Tsuchiya
    •  & Yukihiko Sugimoto
  5. Laboratory of Adjuvant Innovation, National Institute of Biomedical Innovation, Osaka, Japan.

    • Ken J Ishii
  6. Laboratory of Vaccine Science, WPI Immunology Frontier Research Center, Osaka University, Osaka, Japan.

    • Ken J Ishii
  7. Department of Microbiology, Hyogo College of Medicine, Hyogo, Japan.

    • Hiroko Tsutsui
  8. Department of Immunology, Juntendo University School of Medicine, Tokyo, Japan.

    • Hideo Yagita
  9. Research Institute for Biomedical Sciences, Tokyo University of Science, Chiba, Japan.

    • Yoichiro Iwakura
  10. Medical Mycology Research Center, Chiba University, Chiba, Japan.

    • Yoichiro Iwakura
  11. Laboratory for Cytokine Regulation, RIKEN center for Integrative Medical Science, Kanagawa, Japan.

    • Masato Kubo
  12. Division of Molecular Pathology, Research Institute for Biomedical Science, Tokyo University of Science, Chiba, Japan.

    • Masato Kubo
  13. Singapore Immunology Network, Agency for Science, Technology and Research, Biopolis, Singapore.

    • Lai guan Ng
  14. Department of Dermatology, Icahn School of Medicine at Mount Sinai School Medical Center, New York, New York,.

    • Judilyn Fuentes
    •  & Emma Guttman-Yassky

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Contributions

Y.N., G.E. and K.K. designed this study and wrote the manuscript; Y.N., G.E., S. Nakamizo, S.O., S.H., N.K., A.O., A.K., T. Honda and S. Nakajima performed the experiments and analyzed data; S.T. and Y.S. did experiments related to microarray analysis; K.J.I., H.T., H.Y., Y.I., M.K. and L.g.N. developed experimental reagents and gene-targeted mice; J.F. and E.G.-Y. did experiments related to immunohistochemistry of human samples; T.O., T. Hashimoto, Y.M. and K.K. directed the project and edited the manuscript; and all authors reviewed and discussed the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Kenji Kabashima.

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    Supplementary Text and Figures

    Supplementary Figures 1–6 and Supplementary Tables 1–2

Videos

  1. 1.

    Leukocyte cluster formation in elicitation phase of DNFB-induced CHS response

    CMTMR-labeled DNFB-sensitized T cells were transferred into CD11c-YFP mice and then challenged with DNFB to the ear. CD11c+ dermal DCs (green) and T cells (red) formed clusters approximately 6 h after hapten application. The images were taken every 7 min for 24 h.

  2. 2.

    High magnification view of leukocyte cluster in the elicitation phase of CHS

    CMTMR-labeled DNFB-sensitized T cells were transferred into CD11c-YFP mice and then challenged with DNFB to the ear. Sixteen hours later, the established DC–T cell cluster was observed in high magnification view for 2 h every 1 min. In this leukocyte cluster, some of T cells (red) interacted with dermal DCs (green) for more than 2 h. The pale yellow debris are melanin granules. Fragmented red and green debris seems to be indicative of dead T cells and DCs engulfed by macrophages, respectively.

  3. 3.

    T cell division in the skin

    CMTMR-labeled DNFB-sensitized T cells divided in DNFB-challenged site. The mean frequency of T cell division was 1.67±1.81 /h/mm2 (calculated from 5 movies which recorded more than an hour).

  4. 4.

    Macrophages attracted dermal DCs

    TRITC-conjugated dextran was intravenously injected to DNFB-sensitized CD11c-YFP mice to label skin macrophages. The next day, ear skin was challenged with DNFB and examined using two-photon microscopy. In this representative movie, a dermal DC (green) migrated toward TRITC-positive macrophages (red).

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https://doi.org/10.1038/ni.2992

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