Archives of Dermatological Research

, Volume 308, Issue 1, pp 49–54 | Cite as

Japanese Cedar (Cryptomeria japonica) pollen allergen induces elevation of intracellular calcium in human keratinocytes and impairs epidermal barrier function of human skin ex vivo

  • Junichi Kumamoto
  • Moe Tsutsumi
  • Makiko Goto
  • Masaharu Nagayama
  • Mitsuhiro DendaEmail author
Concise Communication


Cry j1 is the major peptide allergen of Japanese cedar (Sugi), Cryptomeria japonica. Since some allergens disrupt epidermal permeability barrier homeostasis, we hypothesized that Cry j1 might have a similar effect. Intracellular calcium level in cultured human keratinocytes was measured with a ratiometric fluorescent probe, Fura-2 AM. Application of Cry j1 significantly increased the intracellular calcium level of keratinocytes, and this increase was inhibited by trypsin inhibitor or a protease-activated receptor 2 (PAR-2) antagonist. We found that Cry j1 itself did not show protease activity, but application of Cry j1 to cultured keratinocytes induced a rapid (within 30 s) and transient increase of protease activity in the medium. This transient increase was blocked by trypsin inhibitor or PAR-2 antagonist. The effect of Cry j1 on transepidermal water loss (TEWL) of cultured human skin was measured in the presence and absence of a trypsin inhibitor and PAR-2 antagonist. Cry j1 significantly impaired the barrier function of human skin ex vivo, and this action was blocked by co-application of trypsin inhibitor or PAR-2 antagonist. Our results suggested that interaction of Cry j1 with epidermal keratinocytes leads to the activation of PAR-2, which induces elevation of intracellular calcium and disruption of barrier function. Blocking the interaction of Cry j1 with epidermal keratinocytes might ameliorate allergic reaction and prevent disruption of epidermal permeability barrier homeostasis.


Cry j1 Protease-activated receptor 2 PAR-2 Trypsin inhibitor FSLLRY-NH2 


Compliance with ethical standards

Conflict of interest

The authors report no conflict of interest.

Supplementary material

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Supplementary material 1 (TIFF 4666 kb)
403_2015_1602_MOESM2_ESM.pdf (2.7 mb)
Supplementary material 2 (PDF 2797 kb)


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Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • Junichi Kumamoto
    • 1
    • 3
  • Moe Tsutsumi
    • 2
  • Makiko Goto
    • 1
    • 2
  • Masaharu Nagayama
    • 1
    • 3
  • Mitsuhiro Denda
    • 1
    • 2
    Email author
  1. 1.Japan Science and Technology Agency, CRESTKawaguchiJapan
  2. 2.Shiseido Research CenterYokohamaJapan
  3. 3.Research Institute for Electronic ScienceHokkaido UniversitySapporoJapan

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