Abstract
Abstract Epidermal Differentiation Complex (EDC) genes are critical for barrier function in the skin, and Th2 cytokines that promote allergic inflammation inhibit EDC gene expression in keratinocytes. To further define the effects of the atopic immune response on keratinocyte differentiation, we investigated IL-4-mediated changes in gene expression in human keratinocytes via RNA-seq. Human keratinocytes were differentiated for 2 days with calcium chloride and then stimulated with IL-4 for 24 hours, or keratinocytes were differentiated with calcium chloride and chronically stimulated with IL-4 for 5 days. At day 2, IL-4 increased expression of genes associated with cell cycle, and decreased expression of genes involved with cell adhesion. At day 5 of differentiation, IL-4-stimulated keratinocytes showed increased expression of genes involved with cell projection and adhesion, and decreased expression of genes that participate in wound repair, epidermis development and host defense. STAT6 inhibition by siRNA significantly altered the expression of less than 20% of genes induced by IL-4. Thus, although IL-4 dramatically alters keratinocyte gene expression and differentiation, only a subset of this response is dependent upon STAT6, and eliminating STAT6 expression has IL-4-independent changes in gene expression. This suggests that keratinocyte gene regulation by IL-4 requires additional molecular pathways.
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