Evaluating IgE Response to DDAC

To further evaluate type of hypersensitivity response induced by DDAC, IgE was evaluated following dermal exposure. Since we previously identified that exposure duration may influence the development of allergic responses (Shane et al., 2017 (link)), IgE levels in mice exposed to DDAC for 14-days were assessed at 28 days post initial exposure. Mice (n=5) were treated for 14-days with DDAC and rested for 14-days. Blood samples were collected via cardiac puncture and sera was then separated by centrifugation and frozen at −20°C for subsequent analysis. To quantify serum IgE levels a standard colorimetric sandwich ELISA was performed according to manufacturer directions (Mouse IgE Ready-Set-Go!, eBioscience). To determine local production of IgE the IgE⁺B220⁺ (IgE⁺ B-cells) population were analyzed by flow cytometry as previously described using IgE-FITC (R35–72) (BD Biosciences) (Shane et al., 2017 (link)).