title: In vivo methods for testing allergenicity show that high hydrostatic pressure hydrolysates of β-lactoglobulin are immunologically inert.
abstract: The major milk allergen β-lactoglobulin (β-LG) exhibits an enhanced susceptibility to proteolysis under high hydrostatic pressure and this may be an efficient method to produce hypoallergenic hydrolysates. The aim of this work was to evaluate the in vivo allergenicity of 3 β-LG hydrolysates produced under atmospheric pressure or high-pressure conditions. Hydrolysates were chosen based on previous experiments that showed that they provide a complete removal of intact β-LG but differed in vitro IgE-binding properties that could be traced to the peptide pattern. The ability to trigger systemic anaphylaxis was assessed using C3H/HeJ mice orally sensitized to β-LG. Outcome measures included symptom score, body temperature, serum mouse mast cell protease 1 (mMCP-1), and quantification of circulating basophils. Mast cell degranulation in vivo was assessed by passive cutaneous anaphylaxis. The 3 tested hydrolysates showed an abrogated allergenicity as revealed by the absence of anaphylactic symptoms and a decrease in body temperature. We demonstrated that the peptides present in the hydrolysates had lost their ability to cross-link 2 human IgE antibodies to induce mast cell degranulation, thus indicating that most of the peptides formed retain just one relevant IgE-binding epitope. The orally sensitized mouse model is a useful tool to address the in vivo allergenicity of novel milk formulas and demonstrates the safety of hydrolysates produced under high-pressure conditions.
authors.affiliation: Department of Bioactivity and Food Analysis, Instituto de Investigación en Ciencias de la Alimentación (CIAL), UAM-CSIC, Madrid, Spain. ivan.lopez@csic.es
authors.forename: I
authors.initials: I
authors.lastname: López-Expósito
authors.forename: R
authors.initials: R
authors.lastname: Chicón
authors.forename: J
authors.initials: J
authors.lastname: Belloque
authors.forename: R
authors.initials: R
authors.lastname: López-Fandiño
authors.forename: M C
authors.initials: MC
authors.lastname: Berin
mesh_terms: D000485:Allergens / Q000276:immunology*; D000707:Anaphylaxis / Q000276:immunology; D000818:Animals; D001491:Basophils / Q000276:immunology; D002417:Cattle; D005260:Female; D006874:Hydrostatic Pressure; D007782:Lactoglobulins / Q000276:immunology*; D051379:Mice; D008809:Mice, Inbred C3H; D011492:Protein Hydrolysates / Q000276:immunology; D059748:Proteolysis
chemical_list: D000485:Allergens; D007782:Lactoglobulins; D011492:Protein Hydrolysates