Identification and partial characterization of multiple major allergens in peanut proteins

Abstract

Peanuts are a major cause of food allergies both in children as in adults which can induce an anaphylactic shock. The identification and characterization of peanut allergens could lead to more insight into the mechanism and contribute to the improvement of diagnostic tests and treatment for peanut allergy. In the present study, the peanut protein-specific immunoglobulin concentrations as well as their recognition of the various peanut proteins or protein subunits was determined in the plasma of peanut-allergic (PA) and non-allergic (NA) individuals. Moreover, two peanut allergens were characterized in more detail to confirm them as the earlier described Ara h1 and Ara h2. The presence of Ig-binding sites in peanut proteins was studied by immunoblotting assays whereas the concentrations of peanut-specific Ig was determined by ELISA. Peanut proteins were found to contain multiple binding sites for immunoglobulins. Of these proteins, six were recognized by peanut-specific IgE present in more than 50% of the plasma samples of the PA group. Their molecular weights were ≈44, 40, 33, 21, 20 and 18 kDa. The last three protein bands were recognized by peanut-specific IgE present in more than 70% of the PA plasma samples and were thought to contain Ara h2. This allergen as well as another protein that was thought to be Ara h1, which was not recognized by the majority of the patients' IgE-containing plasma samples, were isolated and the N terminal amino acid sequence was determined. Peanut protein-specific IgA, IgM, IgG and IgG-subclasses showed a more diverse recognition pattern of peanut protein in the PA group compared to the NA group. No differences were found in the plasma concentrations of peanut protein-specific immunoglobulins of the various classes between the PA and NA group. From the present study, we conclude that peanuts contain multiple allergens, of which six can be described as major allergens, Ara h2 included. In our population Ara h1 is not a major allergen. The recognition of peanut proteins by immunoglobulins is more diverse in PA individuals compared with NA individuals which, however, is not substantiated in the concentrations of peanut-specific immunoglobulins in plasma, other than IgE.