Publication: The Blomia tropicalis allergen Blo t 7 stimulates innate immune signalling pathways through TLR2
2
Issued Date
2018-04-01
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ISSN
13652222
09547894
09547894
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2-s2.0-85042263877
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Mahidol University
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SCOPUS
Bibliographic Citation
Clinical and Experimental Allergy. Vol.48, No.4 (2018), 464-474
Suggested Citation
T. Soongrung, K. Mongkorntanyatip, T. Peepim, A. Buaklin, M. Le Mignon, N. Malainual, E. Nony, A. Jacquet The Blomia tropicalis allergen Blo t 7 stimulates innate immune signalling pathways through TLR2. Clinical and Experimental Allergy. Vol.48, No.4 (2018), 464-474. doi:10.1111/cea.13098 Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/46029
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Title
The Blomia tropicalis allergen Blo t 7 stimulates innate immune signalling pathways through TLR2
Abstract
© 2018 John Wiley & Sons Ltd Background: Although the house dust mite species Blomia tropicalis is a leading cause of allergic diseases in tropical and subtropical regions, the identification and characterization of the allergenic proteins remain incomplete. Objective: We aimed to characterize a recombinant form of Blo t 7 (rBlo t 7) in terms of IgE reactivity, lipid-binding activity and ability to stimulate innate immunity. Methods: The mature Blo t 7 cDNA was cloned by PCR methods for the expression of a secreted form of the allergen in P. pastoris. The IgE reactivity to purified rBlo t 7 as well as the potential cross-reactivity with Der p 7 was determined by ELISA. The lipid-binding capacity of rBlo t 7 was assayed using fluorescent lipid probes. The stimulation of TLR2 signalling pathway by rBlo t 7 was examined in cell activation and reporter assays. Results: The amplified mature Blo t 7 cDNA revealed the presence of a 60 base pair insertion compared with the reference sequence registered in the GenBank database. Multiple protein sequence alignments of group 7 mite allergens confirmed that this longer deduced amino acid sequence was the authentic Blo t 7 polypeptide chain. Analysis of IgE reactivity can classify rBlo t 7 as an intermediate B. tropicalis allergen which displayed weak cross-reactivity with Der p 7. Purified rBlo t 7 was shown to bind selectively the naturally fluorescent lipid probe cis-parinaric (cPNA) with a dissociation constant of 2 μmol/L. The group 7 Blomia allergen stimulated the TLR2-, NF-kB- and MAPK-dependent production of IL-8 and GM-CSF in respiratory epithelial cells. Conclusions & Clinical Relevance: Through its propensity to transport fatty acids/lipids and to stimulate TLR2 signalling pathways in airway epithelial cells, Blo t 7 can represent a key allergen for the initiation of the B. tropicalis-induced airway inflammation.