Satitsuksanoa, P., Kennedy, M., Gilis, D., Le Mignon, M., Suratannon, N., Soh, W. T., Wongpiyabovorn, J., Chatchatee, P., Vangveravong, M., Rerkpattanapipat, T., Sangasapaviliya, A., Piboonpocanun, S., Nony, E., Ruxrungtham, K., Jacquet, A., Thantiworasit, Pattarawat, Pulsawat, Pinya, Daengsuwan, Tassalalpa, Laisuan, Wannada, and Tongdee, Malinee
Background The house dust mite (HDM ) allergen Der p 13 could be a lipid-binding protein able to activate key innate signaling pathways in the initiation of the allergic response. We investigated the IgE reactivity of recombinant Der p 13 (rDer p 13), its lipid-binding activities, and its capacity to stimulate airway epithelium cells. Methods Purified rDer p 13 was characterized by mass spectrometry, circular dichroism, fluorescence-based lipid-binding assays, and in silico structural prediction. IgE-binding activity and allergenic potential of Der p 13 were examined by ELISA, basophil degranulation assays, and in vitro airway epithelial cell activation assays. Results Protein modeling and biophysical analysis indicated that Der p 13 adopts a β-barrel structure with a predominately apolar pocket representing a potential binding site for hydrophobic ligands. Fluorescent lipid-binding assays confirmed that the protein is highly selective for ligands and that it binds a fatty acid with a dissociation constant typical of lipid transporter proteins. The low IgE-binding frequency (7%, n = 224) in Thai HDM-allergic patients as well as the limited propensity to activate basophil degranulation classifies Der p 13 as a minor HDM allergen. Nevertheless, the protein with its presumptively associated lipid(s) triggered the production of IL-8 and GM-CSF in respiratory epithelial cells through a TLR2-, MyD88-, NF-kB-, and MAPK-dependent signaling pathway. Conclusions Although a minor allergen, Der p 13 may, through its lipid-binding capacity, play a role in the initiation of the HDM-allergic response through TLR2 activation. [ABSTRACT FROM AUTHOR]