BACKGROUND: Recombinant hypoallergens that display reduced allergenicity but retain T-cell reactivity represent promising candidates to improve the safety and efficacy of allergen-specific vaccines or immunotherapy. OBJECTIVE: The current study reports the immunologic characterization of a hypoallergenic variant of the major mite allergen Der p 1. METHODS: The recombinant proform of Der p 1 (ProDer p 1) was expressed in Escherichia coli (ProDer p 1 coli), purified and characterized at the level of its secondary structure, and IgE and T-cell reactivities. Moreover, the prophylactic potential of ProDer p 1 coli vaccinations was evaluated in a murine Der p 1 sensitization model. RESULTS: After purification and refolding, ProDer p 1 coli remained aggregated with a higher beta-sheet content and altered Der p 1 conformational epitopes compared with the correctly folded monomeric ProDer p 1 produced in Chinese hamster ovary cells. Both ProDer p 1 forms were able to retain the Der p 1-specific T-cell reactivity but direct ELISA, competitive inhibition, and rat basophil leukemia assays clearly showed that ProDer p 1 coli displays a very weak IgE reactivity. Mice vaccinations with aggregated ProDer p 1 adjuvanted with alum induced a T(H)1-biased immune response that prevented the subsequent allergic response after Der p 1 sensitization and airway challenge with aerosolized mite extracts. Furthermore, ProDer p 1 coli treatment inhibited the development of airway eosinophilia and airway hyperresponsiveness to inhaled methacholine. CONCLUSION: Aggregated forms of Der p 1 could represent hypoallergens suitable for the prevention of mite allergy.
BACKGROUND: Recombinant hypoallergens that display reduced allergenicity but retain T-cell reactivity represent promising candidates to improve the safety and efficacy of allergen-specific vaccines or immunotherapy. OBJECTIVE: The current study reports the immunologic characterization of a hypoallergenic variant of the major mite allergen Der p 1. METHODS: The recombinant proform of Der p 1 (ProDer p 1) was expressed in Escherichia coli (ProDer p 1 coli), purified and characterized at the level of its secondary structure, and IgE and T-cell reactivities. Moreover, the prophylactic potential of ProDer p 1 coli vaccinations was evaluated in a murine Der p 1 sensitization model. RESULTS: After purification and refolding, ProDer p 1 coli remained aggregated with a higher beta-sheet content and altered Der p 1 conformational epitopes compared with the correctly folded monomeric ProDer p 1 produced in Chinese hamster ovary cells. Both ProDer p 1 forms were able to retain the Der p 1-specific T-cell reactivity but direct ELISA, competitive inhibition, and rat basophil leukemia assays clearly showed that ProDer p 1 coli displays a very weak IgE reactivity. Mice vaccinations with aggregated ProDer p 1 adjuvanted with alum induced a T(H)1-biased immune response that prevented the subsequent allergic response after Der p 1 sensitization and airway challenge with aerosolized mite extracts. Furthermore, ProDer p 1 coli treatment inhibited the development of airway eosinophilia and airway hyperresponsiveness to inhaled methacholine. CONCLUSION: Aggregated forms of Der p 1 could represent hypoallergens suitable for the prevention of mite allergy.
Authors: Geoffrey A Mueller; Lori L Edwards; Jim J Aloor; Michael B Fessler; Jill Glesner; Anna Pomés; Martin D Chapman; Robert E London; Lars C Pedersen Journal: J Allergy Clin Immunol Date: 2010-03-11 Impact factor: 10.793
Authors: Christine Y Y Wai; Nicki Y H Leung; Marco H K Ho; Laurel J Gershwin; Shang An Shu; Patrick S C Leung; Ka Hou Chu Journal: PLoS One Date: 2014-11-03 Impact factor: 3.240