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

Yang, Hee Kyu; Kim, Hyo Jung; Oh, Se Jo; Lee, Ki Sun; Lee, Soo Young; Park, Seok Won; Lee, Kyung Eun; Kim, Kyu Earn

Abstract

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.