Abstract
Dog allergy is common worldwide. However, the allergenicity of dog allergy is still unclear in China as well as in special group, such as children. In this study, we chose Can f 6, a major dog allergen which belongs to the lipocalin to study its allergenicity in Chinese dog allergic children. Can f 6 gene was subcloned into pET-28a vector and transformed into E. coli BL21 (DE3) cells for expression. The recombinant Can f 6 was purified by nickel affinity chromatography, identified by SDS-PAGE, and tested for its allergenicity by Western blot with sera and basophil activation test. Secondary structures, B cell epitopes and homology modeling of Can f 6 were predicted by using a series of bioinformatical approaches. And the verification of B cell epitopes was detected by ELISA. The recombinant allergen showed an explicit band with the molecular weight of 20 kDa by SDS-PAGE. Sera from 56.3 % (18/32) of dog-allergic children patients reacted with Can f 6. The induction of the expression of CD63 and CCR3 of dog allergic children in passively sensitized basophils was up to approximately 5.0 times higher than healthy subjects. The secondary structure of Can f 6 contains 3 α-helices, 9 β-sheets and random coils. Five B cell epitopes of Can f 6 were predicted and were confirmed successfully by ELISA. The results showed Can f 6 is a major allergen in Chinese children, which provides a basis for further study of Can f 6 in diagnosis and treatment of symptoms in children in China. The structural information of Can f 6 will help to form a foundation for the future design of vaccines and therapies for Can f 6 related allergies.
Highlights
Dogs are the predominant allergic source worldwide, which can provoke severe allergic disease, such as asthma, urticaria and rhinitis [1]
The codon optimized of Can f 6 sequence was subcloned into pET-28a (+) vector and transformed into BL21 (DE3) pLysS E. coli
The sensitization rate we obtained from this study was 56.3 %, proving that Can f 6 was a major allergen of the dog in Chinese children
Summary
Dogs are the predominant allergic source worldwide, which can provoke severe allergic disease, such as asthma, urticaria and rhinitis [1]. Domesticated dogs initially served as a practical character for protection or agricultural use in human being, but served a more emotional role of companionship recently. With this paradigm shift, dogs begin to move indoors. This unremitting exposure is associated with the progress and deterioration of dog allergic disease. For the patients with dog allergic disease, avoidance is clearly the best treatment modality. This does frequently not occur, even in settings beyond occupational allergy. The sensitization rate to dogs was reported as high as 10 % of all people in Western countries, varying by country, area, time period, and atopic predisposition [2, 3]
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