Experimental immunology
Determination of pea proteins allergenicity with the use Balb/c mouse

One of important criteria in evaluating the allergenicity of proteins in human diet is the resistance of their epitopes to thermal denaturation and hydrolytic processes as affected by digestive enzymes. The effect of the cooking process on the immunogenic properties of pea proteins was determined experimentally on an animal model. Experimental Balb/c mice were fed diets consist protein originated from flour obtained from either raw or cooked pea that was added to diets at different doses. The control group was administered with casein. On termination of the experiment, the animals were weighed, anaesthetized and exsanguinated. The serum obtained was determined with the ELISA method for the level of IgA and IgG antibodies specific to pea proteins. Minimal differences (statistically insignificant) were demonstrated in the level of IgA between particular groups of mice. In contrast, significant differences occurred in the amount of specific IgG. The highest IgG titre was recorded in serum of mouse from the group fed a diet with the addition of flour from raw pea seeds. The immunoblotting assays confirmed those results. A specific reaction was observed to occur between antibodies and protein fractions with molecular mass of ca. 50 kD (vicilin subunits), 70 kD (convicilin) and ca. 60 kD (legumin proteins). Cooking the pea seeds did not influence on the loss of protein immunogenicity. In mice serum there were found specific antibodies primarily to high-molecular subunits of vicilin. This points to strong potential allergenic properties of pea vicilin.