20.500.12556/RUL-147382 Ara h 2-specific IgE epitope-like peptides inhibit the binding of IgE to Ara h 2 and suppress lgE-dependent effector cell activation Background: Clinical and experimental analyses indicate a pathognomonic role for allergen IgE crosslinking through epitope–paratope interactions as a major initial step in the cascade leading to effector cell activation and clinical manifestations of lgE-mediated food allergies. We aimed to undertake the initial development and assessment of Ara h 2-specific IgE epitope-like peptides that can bind to allergenspecific IgE paratopes and suppress effector cell activation. Methods: We performed biopanning, screening, IgE binding, selection and mapping of peptides. We generated synthetic peptides for use in all functional experiments. ImmunoCAP inhibition, basophil and mast cell activation tests, with LAD2 cells, a human mast cell line were performed. Twenty-six children or young adults who had peanut allergy were studied. Results: We identified and selected three linear peptides (DHPRFNRDNDVA, DHPRYGP and DHPRFST), and immunoblot analyses revealed binding to lgE from peanut-allergic individuals. The peptide sequences were aligned to the disordered region corresponding to the loop between helices 2 and 3 of Ara h 2, and conformational mapping showed that the peptides match the surface of Ara h 2 and h 6 but not other peanut allergens. In ImmunoCAP inhibition experiments, the peptides significantly inhibit the binding of IgE to Ara h 2 (p < .001). In basophil and mast cell activation tests, the peptides significantly suppressed Ara h 2-induced effector cell activation (p < .05) and increased the half-maximal Ara h 2 effective concentration (p < .05). Binding of the peptides to specific IgEs did not induce activation of basophils or mast cells. Conclusions: These studies show that the indicated peptides reduce the allergenic activity of Ara h 2 and suppress lgE-dependent basophil and mast cell activation. These observations may suggest a novel therapeutic strategy for food allergy based on epitope–paratop blocking. Ara h 2 ImmunoCAP inhibition basophil activation test epitopes mast cell activation test paratopes peanut allergy peptides specific IgE alergeni Ara h 2 test aktivacije bazofilcev epitopi inhibicija ImmunoCAP test aktivacije mastocitov paratopi alergija na arašide peptidi specifični IgE true false true Angleški jezik Slovenski jezik Članek v reviji 2023-07-03 14:39:03 2023-07-03 14:39:08 2023-11-10 07:31:55 0000-00-00 00:00:00 2023 0 0 Str. 636-647 iss. 6 Vol. 53 2023 0000-00-00 Zaloznikova Objavljeno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 616-056.43 1365-2222 10.1111/cea.14314 151992835 Clin_Experimental_Allergy_-_2023_-_Koro_ec_-_Ara_h_2‐specific_IgE_epitope‐like_peptides_inhibit_the_binding_of_IgE_to_Ara_h.pdf Clin_Experimental_Allergy_-_2023_-_Koro_ec_-_Ara_h_2‐specific_IgE_epitope‐like_peptides_inhibit_the_binding_of_IgE_to_Ara_h.pdf 1 3F2EEA4596BD6AF30245EB392A4B29B7 9f92bfc03574b7029267897656d1649f1c143204ceac20ffeef1ccd8587800ed 8a4dda6b-199e-11ee-a314-00155dcfd717 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=170938 https://onlinelibrary.wiley.com/doi/10.1111/cea.14314 0 611d2f0b-199e-11ee-a314-00155dcfd717 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=170937 Fakulteta za farmacijo 0 0 0