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
Y1 - 2019
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.
UR - https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=85068103712&origin=inward
UR - https://www.ncbi.nlm.nih.gov/pubmed/31207117
U2 - https://doi.org/10.1002/mnfr.201900336
DO - https://doi.org/10.1002/mnfr.201900336
M3 - Article
C2 - 31207117
SN - 1613-4125
JO - Molecular nutrition & food research
JF - Molecular nutrition & food research
M1 - 1900336
ER -