Rational Design, Structure-Activity Relationship, and Immunogenicity of Hypoallergenic Pru p 3 Variants

Stephanie Eichhorn, Angelika Hörschläger, Markus Steiner, Josef Laimer, Bettina M Jensen, Serge A Versteeg, Isabel Pablos, Peter Briza, Laurian Jongejan, Neil Rigby, Juan A Asturias, Antonio Portolés, Montserrat Fernandez-Rivas, Nikolaos G Papadopoulos, Adriano Mari, Lars K Poulsen, Peter Lackner, Ronald van Ree, Fatima Ferreira, Gabriele Gadermaier*

*Korrespondierende/r Autor/in für diese Arbeit

Publikation: Beitrag in FachzeitschriftArtikel

Abstract

SCOPE: Allergies to lipid transfer proteins involve severe adverse reactions; thus, effective and sustainable therapies are desired. Previous attempts disrupting disulfide bonds failed to maintain immunogenicity; thus, the aim is to design novel hypoallergenic Pru p 3 variants and evaluate the applicability for treatment of peach allergy.

METHODS AND RESULTS: Pru p 3 proline variant (PV) designed using in silico mutagenesis, cysteine variant (CV), and wild-type Pru p 3 (WT) are purified from Escherichia coli. Variants display homogenous and stable protein conformations with an altered secondary structure in circular dichroism. PV shows enhanced long-term storage capacities compared to CV similar to the highly stable WT. Using sera of 33 peach allergic patients, IgE-binding activity is reduced by 97% (PV) and 71% (CV) compared to WT. Both molecules show strong hypoallergenicity in Pru p 3 ImmunoCAP cross-inhibition and histamine release assays. Immunogenicity of PV is demonstrated with a phosphate-based adjuvant formulation in a mouse model.

CONCLUSIONS: An in silico approach is used to generate a PV without targeting disulfide bonds, T cell epitopes, or previously reported IgE epitopes of Pru p 3. PV is strongly hypoallergenic while structurally stable and immunogenic, thus representing a promising candidate for peach allergen immunotherapy.
OriginalspracheEnglisch
Aufsatznummer1900336
Seiten (von - bis)1-10
FachzeitschriftMolecular Nutrition and Food Research
Jahrgang63
Ausgabenummer18
DOIs
PublikationsstatusVeröffentlicht - Sep 2019

Bibliographische Notiz

© 2019 The Authors. Published by WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Systematik der Wissenschaftszweige 2012

  • 304 Medizinische Biotechnologie
  • 106 Biologie

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