Biological activity of human IgE monoclonal antibodies targeting Der p 2, Fel d 1, Ara h 2 in basophil mediator release assays

Frontiers in Immunology

Immunology

Lorenz Aglas¹  Hannah L. Widauer¹  Glorismer Pena-Castellanos¹  Maria A. Stigler¹  Serge A. Versteeg²  Ronald van Ree²  Scott A. Smith³  Martin D. Chapman⁴  Bryan R. E. Smith⁴  Anna Pomés⁴ 

[1] Department of Biosciences and Medical Biology, University of Salzburg, Salzburg, Austria;Department of Experimental Immunology, Amsterdam University Medical Centers, Amsterdam, Netherlands;Department of Otorhinolaryngology, Amsterdam University Medical Centers, Amsterdam, Netherlands;Department of Medicine, Vanderbilt University Medical Center, Nashville, TN, United States;InBio, Charlottesville, VA, United States;

关键词: human IgE monoclonal antibodies;    mediator release assay;    Der p 2;    Fel d 1;    Ara h 2;    allergy diagnosis;   

DOI  :  10.3389/fimmu.2023.1155613

 received in 2023-01-31, accepted in 2023-04-27,  发布年份 2023

来源: Frontiers

【 摘 要 】

BackgroundHuman Immunoglobulin E monoclonal antibodies (hIgE mAb) are unique tools for investigating IgE responses. Here, the biological activity of hIgE mAb, derived from immortalized B cells harvested from the blood of allergic donors, targeting three allergens (Der p 2, Fel d 1 and Ara h 2) was investigated.MethodsThree Der p 2-, three Fel d 1- and five Ara h 2-specific hIgE mAb produced by human B cell hybridomas, were combined in pairs and used to passively sensitize humanized rat basophilic leukemia cells and compared with sensitization using serum pools. Sensitized cells were stimulated with corresponding allergens (recombinant or purified), allergen extracts or structural homologs, having 40-88% sequence similarity, and compared for mediator (β-hexosaminidase) release.ResultsOne, two and eight pairs of Der p 2-, Fel d 1- and Ara h 2-specific hIgE mAb, respectively, produced significant mediator release (>50%). A minimum hIgE mAb concentration of 15-30 kU/L and a minimum antigen concentration between 0.01-0.1 µg/mL were sufficient to induce a pronounced mediator release. Individual sensitization with one Ara h 2-specific hIgE mAb was able to induce crosslinking independently of a second specific hIgE mAb. Der p 2- and Ara h 2-specific mAb showed a high allergen specificity when compared to homologs. Mediator release from cells sensitized with hIgE mAb was comparable to serum sensitization.ConclusionThe biological activity of hIgE mAb reported here provides the foundation for novel methods of standardization and quality control of allergen products and for mechanistic studies of IgE-mediated allergic diseases, using hIgE mAb.

【 授权许可】

Unknown   
Copyright © 2023 Pena-Castellanos, Smith, Pomés, Smith, Stigler, Widauer, Versteeg, van Ree, Chapman and Aglas