Identification and characterization of buckwheat allergen / 천식및알레르기

BACKGROUND AND OBJECTIVE: Buckwheat is considered one of the most important food allergens in Korea. Although a very small amount is ingested or inhaled, it can cause serious allergic reactions. However, the major allergens of buckwheat still remain to be elucidated. The aim of our study was to identify and characterize the major allergen of buckwheat seed. MATERIAL AND METHOD: Dialysis membrane with a cut-off MW 1kD was used for the preparation of crude buckwheat seed allergen extract. SDS-PAGE under reducing conditions and IgE immunoblotting were performed using sera from 15 buckwheat sensitive subjects. Isoelectric focusing and lectin blotting assay were done. RESULT: Western blot analysis showed more than 15 IgE-reactive buckwheat proteins. Among them, a 24kD protein was shown to be the most frequently bound to sera from allergic subjects (54%). Isoelectric point of 24kD protein was around 5.9. In lectin blotting assay, 24kD protein did not bind to Con A nor five other lectins. CONCLUSION: A 24kD protein was the most frequently recognized allergenic component in buckwheat seed. Isoelectric point was around 5.9. Glycosylation was not detected in 24kD of buckwheat protein.

Correlation between House Dust Mite Allergen Concentrations in Scalp Dander and Clinical Severity of Atopic Dermatitis in Children / 소아알레르기및호흡기학회지

PURPOSE: There are compelling evidences that house dust mites are important in the pathogenesis of atopic dermatitis. Recently, detection of house dust mite allergens from clothing, skin, and dust from human hair has been reported. To evaluate the importance of house dust mite exposure in the clinical severity of atopic dermatitis. METHODS: We measured house dust mite allergens (Der f1 from Dermatophagoides farinae and Der p1 from D. pteronyssinus) in scalp dander from 27 children with atopic dermatitis and 41 children with asthma only by enzyme-linked immunosorbent assay and examined correlation between house dust mite allergen concentrations in scalp dander and clinical severity of atopic dermatitis in children. RESULTS: Der f1 was detectable in scalp dander samples of 32 patients (47%) from overall 68 patient and Der p1 was detectable in 33 patients (49%). House dust mite allergens (Der f1 or Der p1) were detectable in scalp dander samples of 46 patients (70%) from overall 68 patient. There was no significant differences in house dust mite allergen levels between atopic dermatitis patients and patients with asthma only. There was a tendency of inverse correlation between frequency of scalp washing and concentration of Der f1 in scalp dander extract (r=-0.24, P=0.052). There was a significant correlation between severity grade of atopic dermatitis and concentration of Der f1 in scalp dander (r=0.39, P0.05). CONCLUSION: There was a significant correlation between house dust mite allergen concentrations in scalp dander and clinical severity in children with atopic dermatitis. And these results suggest that exposure to house dust mite allergen is important in the pathogenesis of atopic dermatitis.

Identification and Characterization of German Cockroach Allergen / 소아알레르기및호흡기학회지

PURPOSE: Cockroaches have been reported one of the major causes of allergic disease such as bronchial asthma and allergic rhinitis. About 10% of children and 20% of adult respiratory allergy patients have positive results of allergy skin tests to cockroach allergen. This finding suggests that cockroach is also important allergen in Korea. Among 4,000 species, German cockroach seems predominant species in the Korean peninsula. Identification and charactrerization of the major allergen is the first step in German cockroach allergen research. But, there have been controversies. It may depends upon different identifying methods. The aim of our study was to identify the major allergen in German cockroach using Westernblot. METHODS: Crude German cockroach allergen extracts with or without protease inhibitor cocktail mixture were prepared, and protein patterns of these two extracts were compared using SDS-PAGE and Coommssie stain. Sera from 32 atopic asthmatic patients with positive allergy skin test against cockroach allergen were used for IgE immunoblot. RESULTS: Coomassie stain after SDS-PAGE demonstrated some discordance between the differently prepared crude allergen extracts. In comparison with the allergen extract which abolished endogenous protease activity, the untreated extract revealed new 32, 55 and 78 kD protein bands and 41 kD protein band was disappeared. About 60% of sera with positive allergy skin test revealed IgE binding bands in Westernblot. Among them, 10, 21, 25, 36 and 52 kD protein band might seemed important German cockroach allergen. CONCLUSION: Finding above may suggest that endogenous protease could denature allergenic proteins in the process of crude cockroach allergen extract. The 10, 21, 25, 36, and 52 kD might be the important allergens in German cockroach.

Production of Recombinant Buckwheat Allergen / 소아알레르기및호흡기학회지

PURPOSE: Buckwheat is one of the most common food allegen in Korea and frequently elicit severe allergic reactions. However, up to now, only few reports on buckwheat allergens have been reproted. The purpose of this study was to isolate mRNA for the production of recombinant buckwheat allergens. METHODS: After the isolation of mRNA from ripening buckwheat seeds, in vitro translation was performed. The proteim patters of in vitro traslate products were identified using SDS-PAGE. RESULTS: A260/A260 ratio total RNA was 1.9 and mRNA was 2.0. In vitro translate products showed towered molecular buckwheat proteins such as 1, 3, and 14kD, while other high molecular weight protein of buckwheat seed were not shown in SDS-PAGE. CONCLUSION: mRNA of buckwheat was purified only from ripening seeds. The composition of mRNA was different according to the ripening periods. It is believed that this finding can give a clue to the basic research of buckwheat allergen.