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

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  • 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
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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.

TidsskriftMolecular Nutrition & Food Research
Udgave nummer18
Sider (fra-til)e1900336
StatusUdgivet - sep. 2019

Bibliografisk note

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

ID: 58997313