Rational design of a hypoallergenic Phl p 7 variant for immunotherapy of polcalcin-sensitized patients

Abstract Polcalcins are important respiratory panallergens, whose IgE-binding capacity depends on the presence of calcium. Since specific immunotherapy is not yet available for the treatment of polcalcin-sensitized patients, we aimed to develop a molecule for efficient and safe immunotherapy. We gen...

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Autores principales:	Marianne Raith, Doris Zach, Linda Sonnleitner, Konrad Woroszylo, Margarete Focke-Tejkl, Herbert Wank, Thorsten Graf, Annette Kuehn, Mariona Pascal, Rosa Maria Muñoz-Cano, Judith Wortmann, Philipp Aschauer, Walter Keller, Simone Braeuer, Walter Goessler, Ines Swoboda
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2019
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/fcb00871019f4927a66904b86062fd11
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spelling	oai:doaj.org-article:fcb00871019f4927a66904b86062fd112021-12-02T15:10:03ZRational design of a hypoallergenic Phl p 7 variant for immunotherapy of polcalcin-sensitized patients10.1038/s41598-019-44208-02045-2322https://doaj.org/article/fcb00871019f4927a66904b86062fd112019-05-01T00:00:00Zhttps://doi.org/10.1038/s41598-019-44208-0https://doaj.org/toc/2045-2322Abstract Polcalcins are important respiratory panallergens, whose IgE-binding capacity depends on the presence of calcium. Since specific immunotherapy is not yet available for the treatment of polcalcin-sensitized patients, we aimed to develop a molecule for efficient and safe immunotherapy. We generated a hypoallergenic variant of the grass pollen polcalcin Phl p 7 by introducing specific point mutations into the allergen’s calcium-binding regions. We thereby followed a mutation strategy that had previously resulted in a hypoallergenic mutant of a calcium-binding food allergen, the major fish allergen parvalbumin. Dot blot assays performed with sera from Phl p 7-sensitized patients showed a drastically reduced IgE reactivity of the Phl p 7 mutant in comparison to wildtype Phl p 7, and basophil activation assays indicated a significantly reduced allergenic activity. Rabbit IgG directed against mutant rPhl p 7 blocked patients’ IgE binding to wildtype Phl p 7, indicating the mutant’s potential applicability for immunotherapy. Mass spectrometry and circular dichroism experiments showed that the mutant had lost the calcium-binding capacity, but still represented a folded protein. In silico analyses revealed that the hypoallergenicity might be due to fewer negative charges on the molecule’s surface and an increased molecular flexibility. We thus generated a hypoallergenic Phl p 7 variant that could be used for immunotherapy of polcalcin-sensitized individuals.Marianne RaithDoris ZachLinda SonnleitnerKonrad WoroszyloMargarete Focke-TejklHerbert WankThorsten GrafAnnette KuehnMariona PascalRosa Maria Muñoz-CanoJudith WortmannPhilipp AschauerWalter KellerSimone BraeuerWalter GoesslerInes SwobodaNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 9, Iss 1, Pp 1-10 (2019)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Marianne Raith Doris Zach Linda Sonnleitner Konrad Woroszylo Margarete Focke-Tejkl Herbert Wank Thorsten Graf Annette Kuehn Mariona Pascal Rosa Maria Muñoz-Cano Judith Wortmann Philipp Aschauer Walter Keller Simone Braeuer Walter Goessler Ines Swoboda Rational design of a hypoallergenic Phl p 7 variant for immunotherapy of polcalcin-sensitized patients
description	Abstract Polcalcins are important respiratory panallergens, whose IgE-binding capacity depends on the presence of calcium. Since specific immunotherapy is not yet available for the treatment of polcalcin-sensitized patients, we aimed to develop a molecule for efficient and safe immunotherapy. We generated a hypoallergenic variant of the grass pollen polcalcin Phl p 7 by introducing specific point mutations into the allergen’s calcium-binding regions. We thereby followed a mutation strategy that had previously resulted in a hypoallergenic mutant of a calcium-binding food allergen, the major fish allergen parvalbumin. Dot blot assays performed with sera from Phl p 7-sensitized patients showed a drastically reduced IgE reactivity of the Phl p 7 mutant in comparison to wildtype Phl p 7, and basophil activation assays indicated a significantly reduced allergenic activity. Rabbit IgG directed against mutant rPhl p 7 blocked patients’ IgE binding to wildtype Phl p 7, indicating the mutant’s potential applicability for immunotherapy. Mass spectrometry and circular dichroism experiments showed that the mutant had lost the calcium-binding capacity, but still represented a folded protein. In silico analyses revealed that the hypoallergenicity might be due to fewer negative charges on the molecule’s surface and an increased molecular flexibility. We thus generated a hypoallergenic Phl p 7 variant that could be used for immunotherapy of polcalcin-sensitized individuals.
format	article
author	Marianne Raith Doris Zach Linda Sonnleitner Konrad Woroszylo Margarete Focke-Tejkl Herbert Wank Thorsten Graf Annette Kuehn Mariona Pascal Rosa Maria Muñoz-Cano Judith Wortmann Philipp Aschauer Walter Keller Simone Braeuer Walter Goessler Ines Swoboda
author_facet	Marianne Raith Doris Zach Linda Sonnleitner Konrad Woroszylo Margarete Focke-Tejkl Herbert Wank Thorsten Graf Annette Kuehn Mariona Pascal Rosa Maria Muñoz-Cano Judith Wortmann Philipp Aschauer Walter Keller Simone Braeuer Walter Goessler Ines Swoboda
author_sort	Marianne Raith
title	Rational design of a hypoallergenic Phl p 7 variant for immunotherapy of polcalcin-sensitized patients
title_short	Rational design of a hypoallergenic Phl p 7 variant for immunotherapy of polcalcin-sensitized patients
title_full	Rational design of a hypoallergenic Phl p 7 variant for immunotherapy of polcalcin-sensitized patients
title_fullStr	Rational design of a hypoallergenic Phl p 7 variant for immunotherapy of polcalcin-sensitized patients
title_full_unstemmed	Rational design of a hypoallergenic Phl p 7 variant for immunotherapy of polcalcin-sensitized patients
title_sort	rational design of a hypoallergenic phl p 7 variant for immunotherapy of polcalcin-sensitized patients
publisher	Nature Portfolio
publishDate	2019
url	https://doaj.org/article/fcb00871019f4927a66904b86062fd11
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Rational design of a hypoallergenic Phl p 7 variant for immunotherapy of polcalcin-sensitized patients

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