Complementary methods for contact hypersensitivity (CHS) evaluation in mice.

Contact hypersensitivity (CHS) is an experimental model of allergic contact dermatitis (ACD) that can be studied in mice. For CHS responses, mice are immunized by painting with a reactive hapten, such as 1-fluoro-4,6-dinitrobenzene (DNFB), on the shaved abdominal and chest skin. Subsequently, the ears are challenged with diluted hapten, eliciting 'hypersensitive' ear-swelling reactions, which can be measured with a micrometer. In this manuscript we present complementary methods that can be used to evaluate CHS in mice that include: ear weight, vascular permeability, myeloperoxidase (MPO) activity, IFN-γ concentration in ear extracts and also IFN-γ production by auricular lymph node cells (ELNC). The biochemical evaluation of CHS can be also supported by proliferation assay, measurement of IFN-γ production by skin-draining lymph node cells employing ELISA test and by evaluation of IFN-γ(+) TCRαβ CD8 cells with the use of flow cytometry.