Fig. 3. Experimental design, clinical symptoms and histamine levels. (A) Schematic drawing of the experimental protocol: BALB/c mice (n=8 per group) were subjected to weekly intragastric sensitization with cholera toxin and CMP from day 0 through 35. Challenge was performed at day 45 by intragastric (CMP or SP) (n=7 per group) or sublingual (β-Lg, P34 or OVA) (n=5 per group) protein administration. Control mice received only CMP and were then orally challenged. (B) Symptoms were observed 30 minutes following challenge with CMP or SP and scored according to Table 1. (C) Quantification by ELISA of histamine following oral challenge (mean values±SEM). (D) Cutaneous test: sensitized and control mice were subcutaneously injected with CMP, SP and saline (n=4 per group). The presence of blue color in the skin within minutes after injection was considered a positive cutaneous test. Statistically significant difference by ANOVA: ^***P<0.005. CMP, cow's milk proteins; SP, soy proteins; CT, cholera toxin.