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In search of a vaccine for mouse allergy: significant reduction of Mus m 1 allergenicity by structure-guided single-point mutations.
- Source :
-
International archives of allergy and immunology [Int Arch Allergy Immunol] 2012; Vol. 157 (3), pp. 226-37. Date of Electronic Publication: 2011 Oct 27. - Publication Year :
- 2012
-
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.<br />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.<br />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.<br />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.<br /> (Copyright © 2011 S. Karger AG, Basel.)
- Subjects :
- Adult
Allergens chemistry
Allergens immunology
Animals
Basophil Degranulation Test
Blotting, Western
Circular Dichroism
Epitopes, T-Lymphocyte immunology
Humans
Hypersensitivity diagnosis
Mice
Protein Structure, Secondary
Spectrometry, Fluorescence
Vaccines, Synthetic
Allergens genetics
Hypersensitivity immunology
Immunoglobulin E immunology
Immunotherapy
Mutagenesis, Site-Directed
Point Mutation
T-Lymphocytes immunology
Subjects
Details
- Language :
- English
- ISSN :
- 1423-0097
- Volume :
- 157
- Issue :
- 3
- Database :
- MEDLINE
- Journal :
- International archives of allergy and immunology
- Publication Type :
- Academic Journal
- Accession number :
- 22041937
- Full Text :
- https://doi.org/10.1159/000327551