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

doi:10.1002/mnfr.201900336

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

Department of Clinical Medicine

5 Citations (Scopus)

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.

Original language	English
Journal	Molecular Nutrition & Food Research
Pages (from-to)	e1900336
ISSN	1613-4125
DOIs	https://doi.org/10.1002/mnfr.201900336
Publication status	Published - 1 Sept 2019

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Eichhorn, S., Hörschläger, A., Steiner, M., Laimer, J., Jensen, B. M., Versteeg, S. A., Pablos, I., Briza, P., Jongejan, L., Rigby, N., Asturias, J. A., Portolés, A., Fernandez-Rivas, M., Papadopoulos, N. G., Mari, A., Poulsen, L. K., Lackner, P., van Ree, R., Ferreira, F., & Gadermaier, G. (2019). Rational Design, Structure-Activity Relationship, and Immunogenicity of Hypoallergenic Pru p 3 Variants. Molecular Nutrition & Food Research, e1900336. https://doi.org/10.1002/mnfr.201900336

Eichhorn, S, Hörschläger, A, Steiner, M, Laimer, J, Jensen, BM, Versteeg, SA, Pablos, I, Briza, P, Jongejan, L, Rigby, N, Asturias, JA, Portolés, A, Fernandez-Rivas, M, Papadopoulos, NG, Mari, A, Poulsen, LK, Lackner, P, van Ree, R, Ferreira, F & Gadermaier, G 2019, 'Rational Design, Structure-Activity Relationship, and Immunogenicity of Hypoallergenic Pru p 3 Variants', Molecular Nutrition & Food Research, pp. e1900336. https://doi.org/10.1002/mnfr.201900336

@article{663a92cad31642708fdcdb920f882b07,

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

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.",

author = "Stephanie Eichhorn and Angelika H{\"o}rschl{\"a}ger and Markus Steiner and Josef Laimer and Jensen, {Bettina M} and Versteeg, {Serge A} and Isabel Pablos and Peter Briza and Laurian Jongejan and Neil Rigby and Asturias, {Juan A} and Antonio Portol{\'e}s and Montserrat Fernandez-Rivas and Papadopoulos, {Nikolaos G} and Adriano Mari and Poulsen, {Lars K} and Peter Lackner and {van Ree}, Ronald and Fatima Ferreira and Gabriele Gadermaier",

note = "{\textcopyright} 2019 The Authors. Published by WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.",

year = "2019",

month = sep,

day = "1",

doi = "10.1002/mnfr.201900336",

language = "English",

pages = "e1900336",

journal = "Molecular Nutrition & Food Research",

issn = "1613-4125",

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TY - JOUR

T1 - Rational Design, Structure-Activity Relationship, and Immunogenicity of Hypoallergenic Pru p 3 Variants

AU - Eichhorn, Stephanie

AU - Hörschläger, Angelika

AU - Steiner, Markus

AU - Laimer, Josef

AU - Jensen, Bettina M

AU - Versteeg, Serge A

AU - Pablos, Isabel

AU - Briza, Peter

AU - Jongejan, Laurian

AU - Rigby, Neil

AU - Asturias, Juan A

AU - Portolés, Antonio

AU - Fernandez-Rivas, Montserrat

AU - Papadopoulos, Nikolaos G

AU - Mari, Adriano

AU - Poulsen, Lars K

AU - Lackner, Peter

AU - van Ree, Ronald

AU - Ferreira, Fatima

AU - Gadermaier, Gabriele

PY - 2019/9/1

Y1 - 2019/9/1

N2 - 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.

AB - 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.

U2 - 10.1002/mnfr.201900336

DO - 10.1002/mnfr.201900336

M3 - Journal article

C2 - 31207117

SN - 1613-4125

SP - e1900336

JO - Molecular Nutrition & Food Research

JF - Molecular Nutrition & Food Research

ER -

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

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