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  • The Immune System Response to 15-kDa Barley Protein: A Mouse Model Study.

The Immune System Response to 15-kDa Barley Protein: A Mouse Model Study.

Nutrients (2022-10-28)
Barbara Wróblewska, Ewa Kubicka, Ewelina Semenowicz, Anna Ogrodowczyk, Anita Mikołajczyk, Dagmara Złotkowska
ABSTRACT

Barley (Hordeum vulgare L.) proteins are taxonomically homologous to wheat proteins and react with sera from patients with baker's asthma. In the current work, the crude extract of barley proteins was divided into six fractions on DEAE-Sepharose. Their immunoreactivity in reacting with sera from patients with a confirmed food allergy varied, and the 15-kDa fraction (B-FrVI) showed the strongest response. In silico analysis confirmed that 15-kDa B-FrVI protein belongs to the trypsin/amylase inhibitor family and to a group of MHC type II allergens. In the next step, the immunogenicity of the B-FrVI was examined in a mouse model. It was shown that, compared to the PBS group, administration of B-FrVI to mice induced almost 2× higher amounts of specific IgG, ~217, and IgA ~29, as early as day 28 after immunization, regardless of the route (intraperitoneal or oral) of antigen administration (p < 0.0001). An ELISpot for B-cell responses confirmed it. Stimulation of mesenteric lymphocytes with pure B-FrVI significantly increased (p < 0.001) the proliferation of lymphocytes from all groups compared to cells growing in media only and stimulated with lyophilized beer. The experiments prove the strong immunogenicity of the 15-kDa B-FrVI protein and provide a basis for future studies of the allergenic nature of this protein.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Anti-Mouse IgA (α-chain specific)−Peroxidase antibody produced in goat, affinity isolated antibody, buffered aqueous solution
Sigma-Aldrich
Anti-Mouse IgG (whole molecule) antibody produced in goat, whole antiserum