title: Targeting a cross-reactive Gly m 5 soy peptide as responsible for hypersensitivity reactions in a milk allergy mouse model.
abstract: BACKGROUND
  Cross-reactivity between soybean allergens and bovine caseins has been previously reported. In this study we aimed to map epitopes of the major soybean allergen Gly m 5 that are co-recognized by casein specific antibodies, and to identify a peptide responsible for the cross-reactivity.
  
  
  METHODS
  Cow's milk protein (CMP)-specific antibodies were used in different immunoassays (immunoblotting, ELISA, ELISA inhibition test) to evaluate the in vitro recognition of soybean proteins (SP). Recombinant Gly m 5 (α), a truncated fragment containing the C-terminal domain (α-T) and peptides of α-T were obtained and epitope mapping was performed with an overlapping peptide assay. Bioinformatics tools were used for epitope prediction by sequence alignment, and for modelling the cross-recognized soy proteins and peptides. The binding of SP to a monoclonal antibody was studied by surface Plasmon resonance (SPR). Finally, the in vivo cross-recognition of SP was assessed in a mouse model of milk allergy.
  
  
  RESULTS
  Both α and α-T reacted with the different CMP-specific antibodies. α-T contains IgG and IgE epitopes in several peptides, particularly in the peptide named PA. Besides, we found similar values of association and dissociation constants between the α-casein specific mAb and the different milk and soy components. The food allergy mouse model showed that SP and PA contain the cross-reactive B and T epitopes, which triggered hypersensitivity reactions and a Th2-mediated response on CMP-sensitized mice.
  
  
  CONCLUSIONS
  Gly m 5 is a cross-reactive soy allergen and the α-T portion of the molecule contains IgG and IgE immunodominant epitopes, confined to PA, a region with enough conformation to be bound by antibodies. These findings contribute to explain the intolerance to SP observed in IgE-mediated CMA patients, primarily not sensitised to SP, as well as it sets the basis to propose a mucosal immunotherapy for milk allergy using this soy peptide.
authors.affiliation: Laboratorio de Investigaciones del Sistema Inmune-LISIN, Departamento de Ciencias Biológicas, Facultad de Ciencias Exactas, Universidad Nacional de La Plata, La Plata, Buenos Aires, Argentina ; Centro de Investigaciones y Desarrollo en Criotecnología de Alimentos-CIDCA, Facultad de Ciencias Exactas, Universidad Nacional de La Plata, La Plata, Buenos Aires, Argentina.
authors.forename: Renata
authors.initials: R
authors.lastname: Curciarello
authors.affiliation: Laboratorio de Investigaciones del Sistema Inmune-LISIN, Departamento de Ciencias Biológicas, Facultad de Ciencias Exactas, Universidad Nacional de La Plata, La Plata, Buenos Aires, Argentina.
authors.forename: Paola L
authors.initials: PL
authors.lastname: Smaldini
authors.affiliation: Laboratorio de Investigaciones del Sistema Inmune-LISIN, Departamento de Ciencias Biológicas, Facultad de Ciencias Exactas, Universidad Nacional de La Plata, La Plata, Buenos Aires, Argentina ; Centro de Investigaciones y Desarrollo en Criotecnología de Alimentos-CIDCA, Facultad de Ciencias Exactas, Universidad Nacional de La Plata, La Plata, Buenos Aires, Argentina.
authors.forename: Angela M
authors.initials: AM
authors.lastname: Candreva
authors.affiliation: Centro de Estudios e Investigaciones, Departamento de Ciencias y Tecnología Universidad Nacional de Quilmes, Quilmes, Buenos Aires, Argentina.
authors.forename: Virginia
authors.initials: V
authors.lastname: González
authors.affiliation: Centro de Estudios e Investigaciones, Departamento de Ciencias y Tecnología Universidad Nacional de Quilmes, Quilmes, Buenos Aires, Argentina.
authors.forename: Gustavo
authors.initials: G
authors.lastname: Parisi
authors.affiliation: Centro de Investigación y Desarrollo en Fermentaciones Industriales, CINDEFI (UNLP y CONICET) and Instituto de Investigaciones Bioquímicas de Buenos Aires (IIBBA-CONICET), Fundación Instituto Leloir, Buenos Aires, Argentina.
authors.forename: Ana
authors.initials: A
authors.lastname: Cauerhff
authors.affiliation: Cátedra de Química Medicinal, Departamento de Ciencias Biológicas, Facultad de Ciencias Exactas, Universidad Nacional de La Plata, La Plata, Buenos Aires, Argentina.
authors.forename: Ivana
authors.initials: I
authors.lastname: Barrios
authors.affiliation: Cátedra de Química Medicinal, Departamento de Ciencias Biológicas, Facultad de Ciencias Exactas, Universidad Nacional de La Plata, La Plata, Buenos Aires, Argentina.
authors.forename: Luis Bruno
authors.initials: LB
authors.lastname: Blanch
authors.affiliation: Laboratorio de Investigaciones del Sistema Inmune-LISIN, Departamento de Ciencias Biológicas, Facultad de Ciencias Exactas, Universidad Nacional de La Plata, La Plata, Buenos Aires, Argentina.
authors.forename: Carlos A
authors.initials: CA
authors.lastname: Fossati
authors.affiliation: Centro de Investigaciones y Desarrollo en Criotecnología de Alimentos-CIDCA, Facultad de Ciencias Exactas, Universidad Nacional de La Plata, La Plata, Buenos Aires, Argentina.
authors.forename: Silvana
authors.initials: S
authors.lastname: Petruccelli
authors.affiliation: Laboratorio de Investigaciones del Sistema Inmune-LISIN, Departamento de Ciencias Biológicas, Facultad de Ciencias Exactas, Universidad Nacional de La Plata, La Plata, Buenos Aires, Argentina ; Centro de Investigaciones y Desarrollo en Criotecnología de Alimentos-CIDCA, Facultad de Ciencias Exactas, Universidad Nacional de La Plata, La Plata, Buenos Aires, Argentina.
authors.forename: Guillermo H
authors.initials: GH
authors.lastname: Docena
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000918:Antibody Specificity / Q000276:immunology; D052179:Antigens, Plant / Q000737:chemistry / Q000276:immunology; D002417:Cattle; D003198:Computer Simulation; D003429:Cross Reactions / Q000276:immunology*; D004195:Disease Models, Animal; D004797:Enzyme-Linked Immunosorbent Assay; D018604:Epitope Mapping; D000939:Epitopes / Q000737:chemistry / Q000276:immunology; D005916:Globulins / Q000737:chemistry / Q000276:immunology; D007150:Immunohistochemistry; D007700:Kinetics; D051379:Mice; D016269:Milk Hypersensitivity / Q000276:immunology*; D008894:Milk Proteins / Q000276:immunology; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000276:immunology; D010455:Peptides / Q000276:immunology*; D021122:Protein Subunits / Q000276:immunology; D011994:Recombinant Proteins / Q000276:immunology; D055314:Seed Storage Proteins / Q000737:chemistry / Q000276:immunology; D030262:Soybean Proteins / Q000737:chemistry / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D052179:Antigens, Plant; D000939:Epitopes; D005916:Globulins; D008894:Milk Proteins; D010446:Peptide Fragments; D010455:Peptides; D021122:Protein Subunits; D011994:Recombinant Proteins; D055314:Seed Storage Proteins; D030262:Soybean Proteins; C013894:beta-conglycinin protein, Glycine max