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- Title
Canis familiaris allergen Can f 7: Expression, purification and analysis of B cell epitopes in Chinese children with dog allergies.
- Authors
Wang, Rui-Qi; Wang, Yu-Jie; Xu, Zhi-Qiang; Zhou, Yan-Jun; Cao, Meng-Da; Zhu, Wei; Sun, Jin-Lyu; Wei, Ji-Fu
- Abstract
Dogs are a major source of indoor allergens. However, the prevalence of dog allergies in China remains unclear, especially in children. In the present study, Can f 7, a canine allergen belonging to the Niemann pick type C2 protein family, was selected to study its sensitization rate in Chinese children with dog allergies. The Can f 7 gene was subcloned into a pET-28a vector and expressed in Escherichia coli BL21 (DE3) cells. Recombinant Can f 7 was purified by nickel affinity chromatography, identified by SDS-PAGE electrophoresis, and had its allergenicity assessed by western blot, ELISA and basophil activation tests. Through a series of bioinformatical approaches, B-cell epitopes, secondary structures, and 3 dimensional (3D) homology modeling of Can f 7 were predicted. The activity of the B cell epitopes was verified by ELISA. The recombinant Can f 7 showed a distinct band with a molecular weight of 14 kDa. Six of 20 sera from dog-allergic children reacted positively to the Can f 7. Can f 7 induced an ~4.0-fold increase in cluster of differentiation 63 and C-C motif chemokine receptor R3 expression in basophils sensitized with the serum of dog-allergic children compared with those of non-allergic controls. The secondary structure analysis showed that Can f 7 contains 6 β-sheets. Five B cell epitopes of Can f 7 were predicted, and two of these were confirmed by ELISA. These results indicate that Can f 7 is an important canine allergen in Chinese children and provide novel data for further research concerning the use of Can f 7 in the diagnosis and treatment of Chinese children with canine allergy symptoms.
- Publication
International Journal of Molecular Medicine, 2019, Vol 43, Issue 3, p1531
- ISSN
1107-3756
- Publication type
Article