In search of a vaccine for mouse allergy: Significant reduction of mus m 1 allergenicity by structure-guided single-point mutations

Elena Ferrari, Daniela Breda, Renato Longhi, Luca Vangelista, Clóvis R. Nakaie, Lisa Elviri, Emanuela Casali, Thelma A. Pertinhez, Alberto Spisni, Samuele E. Burastero

Research output: Contribution to journalArticle

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 immuno- genicity, 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.

Original languageEnglish
Pages (from-to)226-237
Number of pages12
JournalInternational Archives of Allergy and Immunology
Volume157
Issue number3
DOIs
Publication statusPublished - Feb 2012

Keywords

  • Allergen mutants
  • Allergenicity
  • Allergens
  • Allergoid
  • Basophil activation test
  • Lipocalin
  • Recombinant allergens
  • Recombinant hypoallergenic allergens
  • T cell epitopes

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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