Sheriene Moussa Afify1,2,3, Andreas Regner1, Luis F Pacios4, Bart R Blokhuis5, Sebastian A Jensen2, Frank A Redegeld5, Isabella Pali-Schöll1,2, Karin Hufnagl1, Rodolfo Bianchini1, Sonja Guethoff6,7, Matthias F Kramer6,7, Alessandro Fiocchi8, Zdenek Dvorak9, Erika Jensen-Jarolim1,2,10, Franziska Roth-Walter1,2. 1. Comparative Medicine, The Interuniversity Messerli Research Institute of the University of Veterinary Medicine Vienna, Medical University Vienna and University of Vienna, Vienna, Austria. 2. Institute of Pathophysiology and Allergy Research, Center of Pathophysiology, Infectiology and Immunology, Medical University of Vienna, Vienna, Austria. 3. Laboratory Medicine and Immunology Department, Faculty of Medicine, Menoufia University, Shibin El Kom, Egypt. 4. Biotechnology Department, ETSIAAB, Center for Plant Biotechnology and Genomics, CBGP (UPM-INIA), Technical University of Madrid, Madrid, Spain. 5. Division of Pharmacology, Faculty of Science, Utrecht Institute for Pharmaceutical Sciences, Utrecht University, Utrecht, The Netherlands. 6. Bencard Allergie GmbH, Munich, Germany. 7. Allergy Therapeutics, Worthing, UK. 8. Childrens Hospital Bambino Gesù, Rome, Italy. 9. Department of Cell Biology and Genetics, Faculty of Science, Palacky University, Olomouc, Czech Republic. 10. Biomedical International R+D GmbH, Vienna, Austria.
Abstract
BACKGROUND: Previously, the protective farm effect was imitated using the whey protein beta-lactoglobulin (BLG) that is spiked with iron-flavonoid complexes. Here, we formulated for clinical translation a lozenge as food for special medical purposes (FSMP) using catechin-iron complexes as ligands for BLG. The lozenge was tested in vitro and in a therapeutical BALB/c mice model. METHODS: Binding of iron-catechin into BLG was confirmed by spectroscopy and docking calculations. Serum IgE binding of children allergic or tolerating milk was assessed to loaded (holo-) versus empty (apo-) BLG and for human mast cell degranulation. BLG and Bet v 1 double-sensitized mice were orally treated with the holoBLG or placebo lozenge, and immunologically analysed after systemic allergen challenge. Human PBMCs of pollen allergic subjects were flow cytometrically assessed after stimulation with apoBLG or holoBLG using catechin-iron complexes as ligands. RESULTS: One major IgE and T cell epitope were masked by catechin-iron complexes, which impaired IgE binding of milk-allergic children and degranulation of mast cells. In mice, only supplementation with the holoBLG lozenge reduced clinical reactivity to BLG and Bet v 1, promoted Tregs, and suppressed antigen presentation. In allergic subjects, stimulation of PBMCs with holoBLG led to a significant increase of intracellular iron in circulating CD14+ cells with significantly lower expression of HLADR and CD86 compared to their stimulation with apoBLG. CONCLUSION: The FSMP lozenge targeted antigen presenting cells and dampened immune activation in human immune cells and allergic mice in an antigen-non-specific manner, thereby conferring immune resilience against allergic symptoms.
BACKGROUND: Previously, the protective farm effect was imitated using the whey protein beta-lactoglobulin (BLG) that is spiked with iron-flavonoid complexes. Here, we formulated for clinical translation a lozenge as food for special medical purposes (FSMP) using catechin-iron complexes as ligands for BLG. The lozenge was tested in vitro and in a therapeutical BALB/c mice model. METHODS: Binding of iron-catechin into BLG was confirmed by spectroscopy and docking calculations. Serum IgE binding of children allergic or tolerating milk was assessed to loaded (holo-) versus empty (apo-) BLG and for human mast cell degranulation. BLG and Bet v 1 double-sensitized mice were orally treated with the holoBLG or placebo lozenge, and immunologically analysed after systemic allergen challenge. Human PBMCs of pollen allergic subjects were flow cytometrically assessed after stimulation with apoBLG or holoBLG using catechin-iron complexes as ligands. RESULTS: One major IgE and T cell epitope were masked by catechin-iron complexes, which impaired IgE binding of milk-allergic children and degranulation of mast cells. In mice, only supplementation with the holoBLG lozenge reduced clinical reactivity to BLG and Bet v 1, promoted Tregs, and suppressed antigen presentation. In allergic subjects, stimulation of PBMCs with holoBLG led to a significant increase of intracellular iron in circulating CD14+ cells with significantly lower expression of HLADR and CD86 compared to their stimulation with apoBLG. CONCLUSION: The FSMP lozenge targeted antigen presenting cells and dampened immune activation in human immune cells and allergic mice in an antigen-non-specific manner, thereby conferring immune resilience against allergic symptoms.
Authors: Isabella Pali-Schöll; Rodolfo Bianchini; Sheriene Moussa Afify; Gerlinde Hofstetter; Simona Winkler; Stella Ahlers; Theresa Altemeier; Hanna Mayerhofer; Karin Hufnagl; Anna D J Korath; Christina Pranger; Raimund Widhalm; Stephan Hann; Thomas Wittek; Anne Kasper-Giebl; Luis F Pacios; Franziska Roth-Walter; Donata Vercelli; Erika von Mutius; Erika Jensen-Jarolim Journal: Clin Transl Allergy Date: 2022-02-12 Impact factor: 5.871
Authors: Sebastian A Jensen; Alessandro Fiocchi; Ton Baars; Galateja Jordakieva; Anna Nowak-Wegrzyn; Isabella Pali-Schöll; Stefano Passanisi; Christina L Pranger; Franziska Roth-Walter; Kristiina Takkinen; Amal H Assa'ad; Carina Venter; Erika Jensen-Jarolim Journal: World Allergy Organ J Date: 2022-09-15 Impact factor: 5.516