Immune response induced after cutaneous applications of BLG or whole milk. Mice received 30 μl of PBS, purified BLG or milk (n = 19-20/group) on intact skin. Serum samples were collected after 6 cutaneous applications and BLG-specific IgE (A) and IgG1 (B) antibodies were quantified on individual serum samples. The numbers of positive mice are indicated above each group. Results are reported in ng/ml derived from standard curves obtained with calibrated anti-BLG purified monoclonal IgG1 or IgE antibodies . Non-specific binding (NSB) was determined using sera from naive mice. A response was considered significant when higher than NSB + 3σ. (C) Systemic specific IL-4, IL-5 and IL-13 secretions were assessed after ex vivo restimulation of splenocytes with BLG.