J Korean Med Sci.  2010 Jun;25(6):829-834. 10.3346/jkms.2010.25.6.829.

Protease Allergens Induce the Expression of IL-25 via Erk and p38 MAPK Pathway

• Yu HS
• Angkasekwinai P
• Chang SH
• Chung Y
• Dong C

• KMID: 2150859
• DOI: http://doi.org/10.3346/jkms.2010.25.6.829

Abstract

Allergic diseases, including asthma, are characterized by T helper type 2 (Th2) cell-mediated inflammations, coupled with tissue infiltration by eosinophils. In this study, we demonstrate that multiple protease allergens, including papain and DerP1, efficiently induce interleukin (IL)-25 and thymic stromal lymphopoietin (TSLP) gene expression, and this phenomenon is dependent on the protease activities of these allergens. The IL-25 cytokine level in bronchial alveolar lavage (BAL) was also profoundly and significantly increased after treatment with papain. Additionally, the levels of Th2 cytokines were significantly increased, as compared to those in the OVA-only treatment group. The various protease allergens triggered the expression of IL-25 and TSLP mRNA in mouse lung epithelial cells (MLE12) and primary mouse lung epithelial cells; these effects were inhibited by the deactivation of the protease activity of papain. The allergen papain activates the ErK and p38 MAP pathways; the inhibition of these pathways, but not the NFkappaB or PI-3 kinase pathways, impairs the induction of IL-25 and TSLP expression by proteases. In this study, we demonstrate that the protease allergens induce IL-25 and TSLP via the MAP kinase signal pathways, and their protease activities are essential to this pathway.

Keyword

L-25; Thymic Stromal Lymphopoietin; Protease Allergen; Mitogen-Activated Protein Kinases

Figure

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  Fig. 1 Papain induced allergic inflammation with significant induction of IL-25. Mice were intranasally treated with papain and OVA (Papain) or OVA only (OVA) for 6 repetitions. (A) Total cell number of bronchoalveolar lavage fluid (BALF). (B) Differential cell counts of BALF. (C) Concentration of cytokines in BALF. (D) Chemokine and cytokine mRNA expression of lung cells (*P<0.05; n=5 mice per group; 3 independent experiments).

• 

  Fig. 2 Protease allergens induced IL-25 and TSLP mRNA expression by lung epithelial cells and fibroblasts. (A) IL-25 and TSLP mRNA expressed by MLE12 cells line after papain stimulation. This expression is inhibited by the protease inhibitor (P. I.), which in this case was a cocktail of protease inhibitors. (B, C) Aspergillus protease (Asp) and DerP1 can also induce the expression of IL-25 and TSLP in mouse embryonic fibroblast (MEF) cells and MLE12 cells (*P<0.05).

• 

  Fig. 3 IL-25 and TSLP gene expression of various cells is induced by protease allergens. (A) IL-25 expression of PLE (primary lung epithelial cell) and (B) IL-25 and TSLP expression of MEF cells are induced by protease stimulations. (C) Additionally, the eotaxin gene expression of MLE12 cells is induced by protease stimulations (*P<0.05).

• 

  Fig. 4 Protease allergens induce IL-25 and TSLP expression via the MAP kinase pathway. (A) Erk, JNK, and p38 MAP kinase pathways were activated via papain treatment. (B) However, boiled papain did not activate the Erk or p38 MAP kinase pathways. (C) The induction of IL-25 and TSLP of MEF cells by papain treatment is inhibited by MAP kinase inhibition. (D) The IL-25 gene expression of MyD88& TIRA-/- MEF after papain treatment is similar to that of wild type MEF. Protease allergen-induced IL-25 expression was not mediated via the MyD88 or TIRAP pathways. I-pI3K, inhibitor of pI3 kinase (LY294002); I-Erk, inhibitor of Erk MAPK (PD98059); I-p38, Inhibitor of p38 MAPK (SB203580); I-JNK, Inhibitor of JNK MAPK (JNK inhibitor II) (*P<0.05).

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