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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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.


  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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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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  1. 1.

    Supplementary Text and Figures

    Supplementary Figures 1–6 and Supplementary Tables 1–2


  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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