Langerhans cells from aryl hydrocarbon receptor deficient mice. Mus musculus

The arylhydrocarbon receptor is a ligand inducible transcription factor. Known to control xenobiotic metabolizing enzymes, it also affects - depending on the cell type - numerous other genes, either directly or indirectly. With respect to the immune system, persistent activation leads to immunosuppression. We asked how the AhR is involved in Langerhans cells. These antigen presenting cells of the skin are responsible for allergies against chemicals (thus xenobiotic metabolism might be relevant) and a recently detected endogenous ligand, FICZ made by UVB radiation from tryptophane, is particularly abundant in the skin. Keywords: comparison gene deficient mouse cells with wild-type Overall design: ear skin epidermal sheets from AhR exon2 deficient mice and wildtype C57BL/6 mice were isolated and cultivated for 16 hours in standard medium. Tissues were then homogenized and Langerhans cells isolated via BSA gradient and cell sorting to greater 95% purity. RNA was preparedand amplified prior to chip-hybridization using the MessageAmp™ Kit of Ambion (Woodward St. Austin, USA). RNA was biotinylated (Enzo Bio Array™HighYield™ RNA transcript labeling kit (Affymetrix, High Wycombe, UK)) and purified. RNA was hybridized to MOE430A gene chips (Affymetrix).