BACKGROUND: Indoor allergens derived from animals and mites often contribute to exacerbation of skin manifestations in atopic dermatitis (AD) patients. OBJECTIVE: To produce and characterize recombinant cat albumin, a cross-reactive animal allergen. METHODS: A complete cDNA coding for cat albumin was obtained by RT-PCR amplification from cat liver RNA. Recombinant cat albumin was expressed in Escherichia coli as hexahistidine-tagged protein, purified by nickel affinity chromatography and studied for IgE reactivity with sera from cat-allergic patients by ELISA and immunoblotting. Furthermore, CD203c expression of basophils from cat-allergic patients upon exposure to recombinant cat albumin was analysed. RESULTS: Recombinant cat albumin, a cross-reactive animal allergen sharing most IgE epitopes with its natural counterpart, was produced in E. coli. It was recognized preferentially by IgE from AD patients and elicited IgE-dependent basophil activation in sensitized patients. CONCLUSIONS: Recombinant cat albumin may be used as a paradigmatic tool to analyse mechanisms of allergen-triggered exacerbation of AD, for diagnostic and, perhaps for therapeutic purposes.
BACKGROUND: Indoor allergens derived from animals and mites often contribute to exacerbation of skin manifestations in atopic dermatitis (AD) patients. OBJECTIVE: To produce and characterize recombinant cat albumin, a cross-reactive animal allergen. METHODS: A complete cDNA coding for cat albumin was obtained by RT-PCR amplification from cat liver RNA. Recombinant cat albumin was expressed in Escherichia coli as hexahistidine-tagged protein, purified by nickel affinity chromatography and studied for IgE reactivity with sera from cat-allergicpatients by ELISA and immunoblotting. Furthermore, CD203c expression of basophils from cat-allergicpatients upon exposure to recombinant cat albumin was analysed. RESULTS: Recombinant cat albumin, a cross-reactive animal allergen sharing most IgE epitopes with its natural counterpart, was produced in E. coli. It was recognized preferentially by IgE from ADpatients and elicited IgE-dependent basophil activation in sensitized patients. CONCLUSIONS: Recombinant cat albumin may be used as a paradigmatic tool to analyse mechanisms of allergen-triggered exacerbation of AD, for diagnostic and, perhaps for therapeutic purposes.
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Authors: Luis Caraballo; Rudolf Valenta; Leonardo Puerta; Anna Pomés; Josefina Zakzuk; Enrique Fernandez-Caldas; Nathalie Acevedo; Mario Sanchez-Borges; Ignacio Ansotegui; Luo Zhang; Marianne van Hage; Eva Fernández; Luisa Arruda; Susanne Vrtala; Mirela Curin; Hans Gronlund; Antonina Karsonova; Jonathan Kilimajer; Ksenja Riabova; Daria Trifonova; Alexander Karaulov Journal: World Allergy Organ J Date: 2020-04-29 Impact factor: 4.084