Modèles animaux d’allergie alimentaire. Application aux protéines de blé

Abstract

The allergens responsible for wheat food allergy are beginning to be characterized. Nevertheless, animal models that produce highly-specific IgE and clinical symptoms comparable to those observed in allergic patients are of great interest for precise studies of allergens and of the mechanisms involved in wheat allergy. The aim of our research was to develop, in the first instance, a mouse model of allergy to a total extract of gliadins, then to use this model to study two major wheat allergens involved in wheat food allergy on children and adults, namely, the lipid-transfer protein (LTP) and the omega-5 gliadins.

Methods

First, we determined the mouse strain and the allergen dose required to induce an optimal allergic reaction to wheat. Three strains of mice (Balb/cJ, B 10.A ND c3 h/hEj) received four successive intraperitoneal injections of a total-gliadin extract (10 or 20 μg) adsorbed on alum. The level of sensitization was determined by assay of gliadin-specific IgE and IgG1 and by the level of cytokines secreted by splenocytes activated in vivo by the gliadins; in vitro basophil (RBL) degranulation tests were also done. In addition, the intensity of the allergic reaction was evaluated in vivo by analysis of the production of type Th2 cytokines and the influx of eosinophils in bronchoalveolar fluid (BAF) after intranasal gliadin extract provocation. Sensitization assays and provocation tests with LTP and omega-5 gliadins were done subsequently.

Results

The highest level of sensitization was observed in the Balb/cJ mice, whatever the dose of gliadins used. After the provocation test, these mice developed an intense-allergic reaction, as demonstrated by the strong production of type Th2 cytokines and the influx of eosinophils in the BAF. In contrast, a weak or no reaction was observed in the other two mouse strains. While sensitization of Balb/cJ mice by omega-5 gliadins was not very effective, administration of LTP induced significant production of specific IgE and IgG1 as well as specific degranulation of RBL cells, but little or no secretion of type Th2 cytokines by activated splenocytes. Induction of an allergic response after intranasal introduction of LTP was not very effective, in spite of the cellular influx in the BAF.

Conclusion

Balb/cJ mice, sensitized by intraperitoneal injection of a total-gliadin extract, appeared to produce some of the markers of wheat allergy, but the results of the sensitization studies with the purified allergens appear to be more complex.