Keratinocyte Tight Junctions (TJs) in the epidermal layer play a crucial role in skin barrier function. An effective immune response in skin is usually initiated by dendritic Antigen Presenting Cells (APCs) in the epidermis and dermis and is finally executed by T lymphocytes. The aim of this study was to investigate whether increased keratinocyte TJ proteins affect immune responses mediated by Immature Dendritic Cells (imDCs). Keratinocytes were treated with the peptide AdhPep-10-2 (10 nM) and then cocultured with DCs for 24 h. Cytokine production was measured by ELISA and a Luminex Multiplexing System. DCs and T cell subsets were analyzed by flow cytometry. Expression of TJ proteins by keratinocytes treated with AdhPep-10-2 increased markedly, whereas CXCL1 chemokine secretion decreased. Coculture of imDCs with AdhPep-10-2-treated keratinocytes inhibited maturation of DCs, reduced production of pro-inflammatory cytokines, and increased the regulatory T (Treg) cell population in the presence of LPS. These results suggest that AdhPep-10-2 peptide increases keratinocyte TJ proteins, thereby inducing anti-inflammatory responses by imDCs under inflammatory conditions. Taken together, these studies show strong possibility that increased keratinocyte TJ proteins play a key role in DC-mediated immunity.