Production of GM-CSF Mediated by Cysteine Protease of Der f in Canine Keratinocytes

Abstract

ABSTRACT. House dust mite (HDM) allergens are the most common allergens for induction of IgE-mediated hypersensitivity. Recently, epicutaneous sensitization with HDM allergens has been emphasized in the development of atopic dermatitis (AD) by producing various soluble factors in keratinocytes. Among the soluble factors, GM-CSF is a key molecule that activates Langerhans cells, antigen-presenting cells in the epidermis. In the present study, we investigated the effects of Dermatophagoides farinae (Der f) on GM-CSF production in a canine keratinocyte cell line, CPEK. CPEKs were found to produce GM-CSF upon stimulation by Der f. The GM-CSF production was suppressed by addition of a cysteine protease inhibitor. The present results suggest that cysteine protease-derived Der f may be an initiator of allergic inflammation by inducing the production of GM-CSF in keratinocytes.

A review of the roles of keratinocyte‐derived cytokines and chemokines in the pathogenesis of atopic dermatitis in humans and dogs

“…93 The GM-CSF production was suppressed by addition of a cysteine protease inhibitor, suggesting that its production was most likely mediated via PAR-2 by Der f 1, which is one of the major allergens and a cysteine protease. 93 Similar to Der f 1, papain, a naturally derived cysteine protease, also induced GM-CSF production with translocation of nuclear factor of activated T cells (NFAT) in CPEK, and production was partially inhibited by ciclosporin. 94 These results suggest that GM-CSF production in CPEK activated via PAR-2 may be regulated not only by NFAT, but also by another transcription factor, such as nuclear factor-kB (NF-kB) or activator protein-1 (AP-1).…”

“…It was also shown that keratinocytes from nonlesional skin of AD subjects exhibited increased spontaneous and PMA‐stimulated production of GM‐CSF . In dogs, CPEK were found to produce GM‐CSF upon stimulation by a house dust mite (HDM) allergen, Dermatophagoides farinae (Der f) . The GM‐CSF production was suppressed by addition of a cysteine protease inhibitor, suggesting that its production was most likely mediated via PAR‐2 by Der f 1, which is one of the major allergens and a cysteine protease .…”

“…In dogs, CPEK were found to produce GM‐CSF upon stimulation by a house dust mite (HDM) allergen, Dermatophagoides farinae (Der f) . The GM‐CSF production was suppressed by addition of a cysteine protease inhibitor, suggesting that its production was most likely mediated via PAR‐2 by Der f 1, which is one of the major allergens and a cysteine protease . Similar to Der f 1, papain, a naturally derived cysteine protease, also induced GM‐CSF production with translocation of nuclear factor of activated T cells (NFAT) in CPEK, and production was partially inhibited by ciclosporin .…”

Involvement of nuclear factor of activated T cells in granulocyte–macrophage colony‐stimulating factor production in canine keratinocytes stimulated with a cysteine protease

“…In canine keratinocytes, Der f 1, a major allergen of Der f, was reported to upregulate mRNA transcription of GM‐CSF . We recently reported that GM‐CSF production in the CPEK cell line was significantly enhanced by stimulation with Der f, which was completely inhibited by a cysteine protease inhibitor . In canine keratinocytes, however, the signalling pathway for GM‐CSF production, mediated by cysteine proteases, has not been clarified.…”

“…16 We recently reported that GM-CSF production in the CPEK cell line was significantly enhanced by stimulation with Der f, which was completely inhibited by a cysteine protease inhibitor. 17 In canine keratinocytes, however, the signalling pathway for GM-CSF production, mediated by cysteine proteases, has not been clarified. In order to develop a molecular targeting therapy, the signalling pathway of GM-CSF production should be clarified.…”