Identification of major rice allergen and their clinical significance in children / 소아과

PURPOSE: Recently, an increase in the number of patients sensitized to rice allergen with or without clinical symptoms has been reported. This study was designed to determine the major allergens in rice and their clinical significance. METHODS: Twenty-four children (15 boys and 9 girls; mean age, 16.3 months) with allergic disease, who were sensitized to rice antigen (by UniCAP) in the Pediatric Allergy Respiratory Center at Soonchunhyang University Hospital, were enrolled in this study. The allergenicity of various types of rice (raw, cooked, and heat-treated, simulated gastric fluid [SGF], and simulated intestinal fluid [SIF]) was investigated using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoglobulin E (IgE) immunoblots. The patients' medical records, including laboratory data and allergy symptoms after ingestion of rice were reviewed. RESULTS: Patients were sensitized to an average of 13.5 food antigens and their mean total IgE was 6,888.7 kU/L. In SDS-PAGE, more than 16 protein bands were observed in the raw rice, whereas only 14-16 kDa and 31-35 kDa protein bands were observed in cooked rice. The common SDS-PAGE protein bands observed in SGF-, SIF-, and heat-treated rice were 9, 14, and 31 kDa. In a heated-rice IgE immunoblot, protein bands of 9, 14, and 31-33 kDa were found in 27.8%, 38.9%, and 38.9% of all sera, respectively, and in 50%, 50%, and 75%, of ser a from the 4 symptomatic patients, respectively. CONCLUSION: The 9-, 14-, and 31-kDa protein bands appeared to be the major allergens responsible for rice allergy symptoms.

Allergen and chow-protein stimulated splenocyte cytokine productions in buckwheat allergic mice / 천식및알레르기

BACKGROUND AND OBJECTIVE: Normal gut immune response to ingestive food proteins is induction of immune tolerance rather than sensitization, even in atopic individuals. Very restricted kinds of food antigens have been known to cause allergic sensitization in humans. To evaluate the differences of systemic T-cell immune responses to sensitized antigen and regular chow-protein, we performed this study. SUBJECTS AND METHODS: Eight naive female, 5-week-old C3H/HeJ mice were grown under the regular mouse chow feeding condition for 4 weeks. During that period, Group I mice were sensitized with buckwheat extract(1mg/dose) mixed with cholera toxin(10 microgram/dose) by intragastric administration at day1, 2, 3, 7, and 21. The sera were obtained at weekly intervals to measure buckwheat-specific and chow-protein-specific IgE, IgG1 and IgG2a antibodies. Splenocyte proliferation assays and cytokine productions were evaluated with buckwheat. chow-protein. and Con A stimulation. Levels of antibodies (IgE, IgG1, IgG2a) and cytokines (IL-4, IL-10, IL-12, INF-gamma were measured by ELISA. RESULTS: The levels of buckwheat specific IgE, IgG1 and IgG2a were markedly increased in Group I mice, but not in Group II mice. Chow-protein specific IgE and IgG1 antibodies were not increased in both groups of mice. The degrees of buckwheat-specific and chow-protein-specific splenocyte proliferations showed low-grade(SI: 6.68 and 4.48. respectively) compared to those by Con A stimulation(SI : 58). Buckwheat stimulated IL-4 productions were markedly increased in Group I mice, which were higher than Con A stimulated production. INF-gamma production was increased in Group I mice by buckwheat stimulation, and in both groups of mice by Con A stimulation. IL-12 production was shown in Con A stimulated culture supernatants in both groups of mice, but in Group I mice with buckwheat stimulation. IL-10 productions were increased in Group I mice with buckwheat, Con A, and chow-protein stimulations, furthermore, markedly increased IL-10 levels were also shown in chow-protein stimulated splenocyte cultures in both groups of mice. CONCLUSION: While Th1 and Th2 immune responses were induced by intragastricly sensitized buckwheat extract, only regulatory immune responses were dominated by regular chow proteins in this system. The minimum ability of chow-protein specific splenocyte proliferation was preserved and IL-10 was the unique cytokine produced by chow-protein simulation.

IgE and IgG Immunoblot Patterns to Three Major Egg Allergens : According to the Routes of Sensitization in Atopic Children(<3 Years Old) / 소아알레르기및호흡기학회지

PURPOSE: Atopic dermatitis is a chronic, inflammatory relapsing skin disease fre- quently seen in children with a history of food allergy. Recently, acute allergic reaction to egg in patients with atopic dermatitis who have never been exposed to egg white protein has been reported. The pattern of antigen-antibody reaction by means of IgE and IgG western blotting was analysed to evaluate the possibility of non- dietary sensitization. METHODS: Total 16 children(<3 years old) with egg allergy were enrolled. Eight patients of them had never ingested egg before(Non-dietary sensitization, NDS) and the other 8 patients had a history of previous egg ingestion without significant clinical symptoms(Dietary sensitization, DS). Egg proteins were analysed by SOS-PAGE and antigen-antibody reaction by means of IgE and IgG was deteded by Western-Blotting. RESULTS: The pattern of IgE antibodies to egg white protein were similar in the two groups. IgG Western blotting to egg white in the two groups, the ovotran sferrine and ovalbumin were significantly bound by 100% of sera, respectively. In DS patients, specific IgG antibodies to ovomucoid was significantly bound by 87% (7/8) of sera whereas only 38%(3/8) of sera had detectable specific IgG antibodies against ovomucoid in NDS patients. CONCLUSION: Theses results suggest that specific IgE to egg white related to exposure of egg white protein regar specific IgG to ovomucoid is related to dietary ingestion of egg. Specific IgG to ovalbumin and ovotrans-ferrine show good clinical correlation, further study might be essential to clarify to this hypothesis.

Age-dependent changes of B cell and T cell-mediated immune responses in naive C3H/HeJ mice under regular mouse chow feeding conditions / 천식및알레르기

BACKGROUND AND OBJECTIVE: Murine system for studying allergic diseases has been popular in the fields of food allergy and development of their therapeutic strategies. However, there has been no information about the age-dependent changes of natural immune responses of naive C3H/HeJ mice. The purpose of this study was to evaluate the age-dependent changes of B and T-cell mediated immunologic parameters in naive C3H/HeJ mice, which can provide information for experimental planning and analysis of research results. SUBJECTS AND METHODS: Eight naive, female, 5-week-old C3H/HeJ mice were grown under the regular mouse chow feeding conditions for 6 weeks. Sera were obtained at week (w) 5, w6, w8 and w10 for measuring total and chow-specific IgE, IgG1 and IgG2a antibodies. Splenocyte proliferation (at w8 and w10) and cytokine production (at w6, w8 and w10) were evaluated with or without Con A stimulation with pooled splenocytes from two mice of each age group. Serum antibodies and cytokines (IL-4, IL-5, IL-12, INF-gamma, TGF-beta1) were measured by ELISA. Using RT-PCR, IL-4 and INF-gamma mRNA expressions were measured in Peyer's patch and spleen tissue at w10. RESULTS: The levels of total IgE and IgG1 were increased by age while the level of IgG2a was decreased. Chow-specific IgE and IgG2a responses were neglectable through out the whole experimental period (20-30 ng/ml or less). Chow-specific IgG1 levels were measured in the significant concentrations (200-300 ng/ml) but there was no age-dependent change through out the experiment. Con A stimulated-splenocyte proliferation indexes were variable according to the culture-durations and ages of mice. The higher proliferation indexes were observed in the wells receiving thymidine pulse at 48-hour culture, especially in the mice at w10. Con A stimulated IL-4 production in the 72-hour splenocyte culture supernatant was significantly increased at w8, and w10 while INF-gamma production increased only at w10. The changes in the production of IL-5, IL-12 and TGF-beta did not provide significant information in the present study. The ratio of IL-4/IFN-gamma mRNA expression was higher in Peyer's patch than in the spleen. CONCLUSION: The changes of B-cell and T-cell mediated immunologic parameters were complex and variable according to the age in naive C3H/HeJ mice under regular chow feeding conditions. For that reason, the information from the present study needs to be considered in the course of planning or analysing research/data using murine systems.