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Updated: Apr 9, 2026

Humanized Mediator Release Assay as a Read-Out for Allergen Potency
Published on: June 29, 2021
Complexes of allergen and specific IgG antibodies activate human neutrophils
Bibiana Aigner1, Gordana Wozniak-Knopp2, Nahla Ibrahim3
1Institute of Pathophysiology and Allergy Research, Center for Pathophysiology, Infectiology and Immunology, Medical University of Vienna, Vienna, Austria.
Aim:
To investigate whether AICs activate neutrophils via FcγR and influence allergen-uptake and presentation.
Methods:
Major birch pollen allergen was incubated with different monoclonal antibodies (mAbs) and their effector-attenuated LALA-variants. AIC formation was analysed by dynamic light scattering. Neutrophils isolated from birch pollen allergic and non-allergic donors were stimulated with GM-CSF and IFN-γ. FcγR expression and internalisation of fluorescence-labelled allergen were determined by flow cytometry. Neutrophils pulsed with allergen, AICs or LALA-AICs were subjected to DNA-release assays and served as APC for autologous allergen-specific T-cells.
Results:
Cytokine-primed neutrophils showed upregulated FcγRI/CD64, downregulated FcγRIII/CD16, and unaltered FcγRII/CD32 expression. AICs with one and two mAbs were generated. Neutrophils phagocytosed larger AICs and LALA-AICs more effectively than smaller complexes and non-complexed allergen. Compared to non-complexed allergen, T-cell proliferation was inconclusive when neutrophils were pulsed with AICs and enhanced when pulsed with LALA-AICs. AICs but not LALA-AICs induced NET-release.
Conclusion:
FcγR-independent phagocytosis of AICs enhanced the allergen-presenting activity of neutrophils but FcγR-mediated NET induction might interfere with the T cell stimulatory properties. Our results suggest a novel link between humoral and cellular responses to allergens.
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