Clinical and experimental allergy : journal of the British Society for Allergy and Clinical Immunology · 2026
Patient-Derived IgG Epitope Mapping of Bet v 1 Reveals Hypoallergenic Peptide Candidates for Safe and Next-Generation Allergen Immunotherapy.
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Šošić Lara, Paolucci Marta, Streuli Alessandro, Melillo David A, Jiang Tianchi, Campana Raffaela, Lang Claudia C V, Kündig Thomas M, Steuer Christian, Eyer Klaus, Johansen Pål
Аннотация
BACKGROUND: Allergen immunotherapy (AIT) is the only curative treatment for allergic diseases, primarily by inducing allergen-neutralising IgG antibodies. However, its use is limited by frequent allergic adverse events.
OBJECTIVE: To facilitate the design of IgG-epitope-based peptide vaccines, this study aimed to identify IgG-binding regions on the major birch pollen allergen, Bet v 1, using patient-derived immune profiles.
METHODS: Blood samples were collected from 30 birch pollen-allergic patients, both AIT-treated and untreated; healthy individuals were included as controls. Sera were analysed for allergen-specific IgE, IgG, and IgG4, and basophil degranulation inhibition was assessed in humanised RBL cells and blood from allergic individuals. We used DropMap microfluidics for single-cell affinity profiling of IgG- and IgG4-secreting B cells. IgG epitopes were mapped using overlapping peptides and CLIPS technology. Candidate AIT peptides were synthesised on solid phase via Fmoc strategy, and their allergenicity was assessed in basophil degranulation and cellular antigen stimulation assays.
RESULTS: Sera from AIT-treated patients exhibited elevated IgG and IgG4 levels and enhanced inhibition of basophil degranulation. Single-cell analysis indicated IgG-affinity maturation with AIT. IgG-epitope mapping identified four distinct, non-overlapping IgG-binding regions on Bet v 1. Six peptides derived from these regions were successfully produced and did not induce basophil degranulation in vitro. The application of DropMap microfluidics allowed high-resolution single B-cell analysis, providing novel insights into allergen-specific B-cell responses and the effects of AIT at the clonal level.
CONCLUSION: This integrated approach identifies hypoallergenic peptides that selectively engage protective IgG responses and offers a framework for developing safer, next-generation immunotherapies for allergic disease.
OBJECTIVE: To facilitate the design of IgG-epitope-based peptide vaccines, this study aimed to identify IgG-binding regions on the major birch pollen allergen, Bet v 1, using patient-derived immune profiles.
METHODS: Blood samples were collected from 30 birch pollen-allergic patients, both AIT-treated and untreated; healthy individuals were included as controls. Sera were analysed for allergen-specific IgE, IgG, and IgG4, and basophil degranulation inhibition was assessed in humanised RBL cells and blood from allergic individuals. We used DropMap microfluidics for single-cell affinity profiling of IgG- and IgG4-secreting B cells. IgG epitopes were mapped using overlapping peptides and CLIPS technology. Candidate AIT peptides were synthesised on solid phase via Fmoc strategy, and their allergenicity was assessed in basophil degranulation and cellular antigen stimulation assays.
RESULTS: Sera from AIT-treated patients exhibited elevated IgG and IgG4 levels and enhanced inhibition of basophil degranulation. Single-cell analysis indicated IgG-affinity maturation with AIT. IgG-epitope mapping identified four distinct, non-overlapping IgG-binding regions on Bet v 1. Six peptides derived from these regions were successfully produced and did not induce basophil degranulation in vitro. The application of DropMap microfluidics allowed high-resolution single B-cell analysis, providing novel insights into allergen-specific B-cell responses and the effects of AIT at the clonal level.
CONCLUSION: This integrated approach identifies hypoallergenic peptides that selectively engage protective IgG responses and offers a framework for developing safer, next-generation immunotherapies for allergic disease.