House dust mite allergen Der f 2 drives IL-6 and GM–CSF expression in airway epithelial cells via p38 MAPK/NF-κB signaling

Abstract

Objective

Der f 2, a major allergen derived from Dermatophagoides farinae, is a leading cause of allergic asthma. IL-6 and GM–CSF play essential roles in the exacerbation of asthma. However, the mechanical act by which Der f 2 mediates the expression of IL-6, IL-8, and GM–CSF in airway epithelial cells remains incompletely elucidated. Herein, we aimed to explore the effect of Der f 2 on IL-6 and GM–CSF expression in the human airway epithelial cell BEAS-2B and A549.

Methods

Recombinant Der f 2 (rDf2) was acquired using Pichia pastoris. BEAS-2B and A549 cells were used as cell model. The expression of genes and proteins and the involvement of the signaling cascade were assessed using RT–PCR, quantitative real-time PCR (qPCR), Western blotting, and ELISA, respectively.

Results

Our findings showed that rDf2 significantly induced mRNA expression and protein production of IL-6 and GM-CSF in BEAS-2B and A549 cells. In contrast, rDf2 did not influence IL-8 expression or production in both cells. Mechanistic studies revealed that rDf2 triggered activation of the p38 MAPK and JNK. Inhibition of p38, but not JNK, significantly attenuated rDf2-induced IL-6 and GM–CSF expression and production.

Conclusion

This study demonstrates that Der f 2 promotes the expression and production of the pro-inflammatory cytokines IL-6 and GM–CSF in airway epithelial cells via activation of the p38 signaling pathway. These findings provide insights into the molecular mechanisms that Der f 2 may exacerbate airway inflammation.

Keywords:

• House dust mite
• Der f 2
• cytokines
• airway epithelial cells
• MAPK
• NF-κB

Declaration of interest

The authors report no conflicts of interest. The authors alone are responsible for the content and writing of this article.

Availability of data and materials

The datasets used and/or analyzed during the present study are available from the corresponding author on reasonable request.

Authors’ contributions

YCL and WCW performed the majority of experiments and analyzed the data; HLL analyzed data and performed statistical analysis; JJT and SHK designed the study, interpreted the results, and confirmed the authenticity of all the raw data; SHK wrote the manuscript and acquired funding. All authors have read and approved the final version of manuscript.

Ethics approval and consent to participate

Not applicable.

Patient consent for publication

Not applicable.

Additional information

Funding

This work was supported by the China Medical University Hospital (grant no. DMR-111-049) and the Chung Shan Medical University (grant no. CSMU-INT-111-01).