In Search of a Vaccine for Mouse Allergy: Significant Reduction of Mus m 1 Allergenicity by Structure-Guided Single-Point Mutations

In Search of a Vaccine for Mouse Allergy: Significant Reduction of Mus m 1 Allergenicity by Structure-Guided Single-Point Mutations

Author Ferrari, Elena Google Scholar
Breda, Daniela Google Scholar
Longhi, Renato Google Scholar
Vangelista, Luca Google Scholar
Nakaie, Clovis Ryuichi Autor UNIFESP Google Scholar
Elviri, Lisa Google Scholar
Casali, Emanuela Google Scholar
Pertinhez, Thelma A. Google Scholar
Spisni, Alberto Google Scholar
Burastero, Samuele E. Google Scholar
Institution Ist Sci San Raffaele
Univ Parma
CNR
Universidade Federal de São Paulo (UNIFESP)
Abstract Background: Mouse urinary proteins are relevant allergens from mice urine. We used the recombinant protein Mus m 1 as an allergen model to identify if, by altering Mus m 1 architecture via single-point mutations, we could effectively modify its allergenicity. Methods: Based on structural considerations, we synthesized two single-point mutants, Mus m 1-Y120A and Mus m 1-Y120F, which were expected to harbor large structural alterations. Circular dichroism and fluorescence analysis showed significant conformational rearrangements of the aromatic side chains in the internal cavity of Mus m 1-Y120A when compared to Mus m 1-Y120F and Mus m 1. Evaluation of the allergenic potential of the recombinant molecules was performed in vitro with both immunochemical approaches and assays based on the measurement of basophil degranulation. Moreover, to assess the integrity of the T cell epitopes and as an in vitro measure of immunogenicity, we tested the reactivity of T lymphocytes from subjects allergic to mouse urine against proteins and synthetic peptides encompassing the immunodominant linear epitope containing the mutation. Results: We found that the selected point mutation was able to modulate the protein allergenicity, and to severely impair the recognition of Mus m 1 by IgE, while T cell reactivity was fully maintained. Conclusions: in silico predicted, minimum selected structural modifications allowed to design one protein with reduced allergenicity and preserved immunogenicity. Structurally guided mutations can direct the design of proteins with reduced allergenicity which can be used as vaccines for a safer and more effective immunotherapy of allergic disorders. Copyright (C) 2011 S. Karger AG, Basel
Keywords Allergen mutants
Allergenicity
Allergens
Allergoid
Lipocalin
Recombinant allergens
Recombinant hypoallergenic allergens
Basophil activation test
T cell epitopes
Language English
Date 2012-01-01
Published in International Archives of Allergy and Immunology. Basel: Karger, v. 157, n. 3, p. 226-237, 2012.
ISSN 1018-2438 (Sherpa/Romeo, impact factor)
Publisher Karger
Extent 226-237
Origin http://dx.doi.org/10.1159/000327551
Access rights Open access Open Access
Type Article
Web of Science ID WOS:000300405800003
URI http://repositorio.unifesp.br/handle/11600/34381

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