4024 Cysteine Protease Allergen Def f 1 Induces Th2 Cytokines in Mouse Bone Marrow Derived Basophils Via ERK and JNK Dependent Pathways

Saturday, 17 October 2015
Hall D1 Foyer (Floor 3) (Coex Convention Center)

Myung-Hee Yi, PhD , Department of Environmental Medical Biology and Institute of Tropical Medicine, Yonsei University College of Medicine, Seoul, South Korea

Kyoung Yong Jeong, PhD , Derparment of Internal Medicine and Institute of Allergy, Yonsei University College of Medicine, Seoul, South Korea

Ju-Yeong Kim, MD , Department of Environmental Medical Biology, Yonsei University College of Medicine, Seoul, South Korea

Tai-Soon Yong, MD, PhD , Department of Environmental Medical Biology and Institute of Tropical Medicine, Yonsei University College of Medicine, Seoul, South Korea

Background

Recent studies have shown that basophils contribute to the initiation of Th2 cytokine–mediated inflammation. However, the underlying mechanism by which protease allergens triggering basophils to produce Th2 cytokine remains unclear.

Objectives

The objective of this study is to demonstrate the role of the mitogen activated kinase (MAPK) family in mouse bone marrow-derived basophils (BMBs) when treated with cysteine protease allergen Der f 1.

Methods BMBs were selected as DX5+FceRI+c-Kit– by flow cytometry of the mouse bone marrow culture. Sorted BMBs were stimulated with proteolytically active recombinant Der f 1 expressed in the yeast Pichia pastoris. The expression levels of Th2 cytokines, IL-4 and IL-13, were examined by real time PCR and ELISA. The activities of various intracellular MAPK signaling components were assessed by FACS.

Results

Production of both IL-4 and IL-13 in mouse basophils was induced by the treatment of enzymatically active Der f 1, whereas inactive Der f 1 did not. Extracellular signal-regulated kinase (ERK) and c-Jun N-terminal kinases (JNK) were phosphorylated upon treatment of active Der f 1 on BMBs. Additionally, treatment of pharmacological inhibitors PD098059 (ERK inhibitor) and SP600125 (JNK inhibitor) were able to inhibit IL-4 and IL-13 secretion. However, p38 MAPK was not inhibited by its inhibitor (SB203580).

Conclusions

These data suggest involvement of ERK, and JNK MAPKs in signal pathway for the production of Th2 cytokines such as IL-4 and IL-13 in BMBs when treated with a cysteine protease allergen rDer f 1.