Therefore, we studied a large panel of immune markers in this model to represent both the IgE- and non-IgE-mediated regulation of wheat allergenicity

Therefore, we studied a large panel of immune markers in this model to represent both the IgE- and non-IgE-mediated regulation of wheat allergenicity. The cytokine IL-10 is regarded as a regulatory and anti-inflammatory cytokine in humans and in some animal models. with SSWP via skin exposure. In the AA model, mice were sensitized by an intraperitoneal injection of SSWP with alum. In both models, allergic reactions were elicited using an identical protocol. Robust IgE as well as mucosal mast cell protein-1 responses were elicited similarly in both models. However, an analysis of the spleen immune markers recognized strikingly different molecular activation patterns in these two models. Furthermore, a number of immune markers associated with intrinsic allergenicity were also recognized in both models. Since the AF model uses skin exposure without an adjuvant, the mechanisms in the AF model may more closely simulate the human wheat allergenicity mechanisms from skin exposure in occupational settings such as in the baking industry. test, 0.05. 2.2. Comparison of Wheat Protein-Induced Elevation of Total Plasma IgE Antibody Levels in Adjuvant-Free vs. Alum-Adjuvant Mouse Models An allergen-induced elevation of plasma total IgE (TIgE) levels is usually reported as a useful marker of allergenicity CGB in mouse models [23,42,43,44,45,47,48,49]. Therefore, we tested this readout in this study using an optimized ELISA. In the AF model, as obvious in Physique 2A, a significant elicitation of TIgE was noted. The control group of mice did not show significant TIgE responses (Physique 2A). Mice that were sensitized using the AA method also showed a significant elevation of TIgE levels (Physique 2B). The alum-alone injected control mice did not show a significant elevation of TIgE levels (Physique 2B). Open in a separate window Physique 2 (A,B). Comparison of wheat protein-elicited plasma total IgE antibody responses in the adjuvant-free vs. the alum-adjuvant mouse models of wheat allergenicity. (A) In the AF model, Balb/c mice were exposed to SSWP Sesamolin once a week for 6 weeks via the transdermal route, as explained in the methods. A group of control mice did not receive this exposure. Plasma collected after 6 weeks of exposure sensitization was used in the TIgE antibody analysis using an ELISA method explained previously [42]. Physique shows the TIgE levels in allergic mice vs. the control mice in the Sesamolin AF model. (B) In the AA model, Balb/c mice were injected with SSWP along with alum by the intraperitoneal route, as explained in the methods. A group of control mice received alum only for the injection. Plasma collected after 6 weeks of sensitization was used in the TIgE antibody analysis using an ELISA method explained previously [32]. Physique shows the TIgE levels in allergic vs. the control mice in the AA model. * Students test, 0.05. 2.3. Comparison of Wheat Sesamolin Protein-Specfic IgG1 Antibody Responses in Adjuvant-Free vs. Alum-Adjuvant Mouse Models In the AF model, the wheat-specific IgG1 (WSIgG1) antibody levels were measured using a highly sensitive ELISA explained by us before [32,42]. As obvious (Physique 3A), a significant elevation of WSIgG1 was noted in the skin-exposed mice but not in the control group (Physique 3A). In the AA model also, a significant elicitation of WSIgG1 was noted (Physique 3B). The alum-alone injected control mice did not show WSIgG1 responses (Physique 3B). Open in a separate window Physique 3 (A,B). Comparison of the wheat protein-specific IgG1 antibody responses in the adjuvant-free vs. the alum-adjuvant mouse models of wheat allergenicity. (A) In the AF model, Balb/c mice were exposed to SSWP once a week for 6 weeks via the transdermal route, as explained in the methods. A group of control mice did not receive this exposure. Plasma collected after 6 weeks of exposure sensitization was used in the WSIgG1 antibody analysis using an ELISA method explained previously [32]. Physique shows the WSIgG1 levels in allergic mice vs. the control mice in the AF model. (B) In the AA model, Balb/c mice were injected with SSWP along with alum by the intraperitoneal route, as explained in the methods. A group of control mice received alum only for the injection. Plasma collected after 6 weeks of sensitization was used in the WSIgG1 antibody analysis using an ELISA method explained previously [32]. Physique shows the WSIgG1 levels in allergic mice vs. the control mice in the AA model. * Students test, 0.05. 2.4. Comparison of Wheat Protein-Specfic IgG2a Antibody Responses in Adjuvant-Free vs. Alum-Adjuvant Mouse Models The food-specific IgG2a antibody response is commonly used as an in vivo biomarker of a Th1 response because of its dependence on the Th1 cytokine IFN-g [23,47]. The wheat-specific IgG2a.