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Although a marked rise in the prevalence of allergic diseases over the past few decades may be related to environmental factors in industrialized countries, evidence for the protective effect of humidity on the barrier function of the skin is still awaited. We asked whether an increase in the water content of stratum corneum at the site of hapten application had a strong impact on the magnitude of contact hypersensitivity (CHS). The magnitude of CHS, induced by either lipid-soluble or water-soluble hapten, was inversely correlated with the water content of stratum corneum at the hapten application site in the elicitation phase. An increase in the water content induced by exposure to high humidity for 6 hours was sufficient to ameliorate the magnitude of CHS even in mice with the genetic defect in attenuating the CHS responses, such as flaky tail mice. The reduced CHS was associated with downregulation of IL-1α, IL-4, and IFN-γ mRNA expression. Epicutaneously applied hapten can penetrate more readily through the stratum corneum with lower water content than that with higher water content, even after tape-stripping. These findings indicate that increased levels of water in the stratum corneum serve to ameliorate the CHS beyond the genetic effects., (Copyright © 2016 The Authors. Published by Elsevier Inc. All rights reserved.)