Studies on some potential allergenic proteins

Abstract

Background: In our nation, the escalating prevalence of respiratory allergies and bronchial asthma is attributable to a combination of factors such as adverse climatic conditions, ambient air pollution, suspended airborne particulate matter, and plant-derived dust. Previous research has explored the causes, risk factors, prevalence, and incidence of asthma and respiratory allergies to some extent. Allergen proteins in plant pollen is widely studied yet there is lack of research results, derived from studies on common street plants in Ulaanbaatar. It is important to determine the biochemical characterization of allergens to compare them with well characterized plant allergens from countries to see if there are regional differences. And studying allergens in traditional Mongolian milk drinks is often neglected. In order to determine allergenic properties, it is important to identify and characterize the potential allergenic proteins in that particular food source, obtained from respective location. Addressing these issues has provided the impetus for conducting this research. Aim: The aim of this study is to investigate potential allergenic proteins in common some plants and traditional fermented mare’s milk beverage in Mongolia. Materials and Methods: The study employed a cross-sectional survey design. Plant pollen samples were collected during the blooming season, and mare’s milk and matured airag (local fermented mare milk beverage) samples were procured for research in the summer of 2023. Previous researches indicated that key allergens typically exhibit similar characteristics: they are water-soluble glycoproteins ranging in size from 10 to 70 kDa. Therefore we optimized our method to extract the potential allergenic proteins in this range. After extraction, total protein content was determined using the Lowry method. SDS-PAGE was conducted to determine molecular mass of extracted proteins, and acute toxicity of the fermented mare milk’s potential allergen solution was assessed on animal model according to standard protocols. Results: The Lowry method revealed that the pollen allergen solution from Bromus inermis contained 1.5±0.02 mg/ml of protein, the white birch pollen solution contained 1.2±0.05 mg/ml, and the magwort (Artemisia macrocephala) allergen solution contained 1.85±0.02 mg/ml proteins. In the White birch pollen solution exhibited 7 distinct bands (18, 26, 32, 55, 66, 84, 97 kDa molecular weights), birch pollen solution had 5 distinct bands (14, 19, 32, 45, 56 kDa), magwort pollen allergen solution had 4 distinct bands (13, 15, 60, 70 kDa), and the fermented mare’s milk allergen solution contained 4 proteins (12, 30, 50, 68 kDa). Acute toxicity studies on the fermented mare’s milk allergen solution, conducted on C57BL/6 mice weighing 22-25 g, revealed no statistically significant differences in the absolute and relative weights of the liver, spleen, kidney, heart, and lungs between the control and experimental groups following oral administration of the yeast allergen solution at doses ranging from 500-5000 mg/kg. The LD50 value for the fermented mare’s milk allergen solution was determined to be greater than 5000 mg/kg. Conclusion: The study successfully isolated potential allergenic proteins from plant pollen and fermented mare’s milk allergens. The acute toxicity assessment of the yeast allergen solution indicated an LD50 value exceeding 5000 mg/kg. In alignment with the toxicity classifications of Hodja and Sterner, as well as OECD guidelines, the yeast allergen solution can be classified as practically non-toxic.