The interaction of B7 family members with appropriate receptors is essential for an effective T cell response. CD80 and CD86 are the principal co-stimulatory molecules of this family and they are mainly expressed on professional antigen presenting cells (APCs), but also on several non-lymphoid cells. CD86 is constitutively expressed in keratinocytes from the spinous layer of normal cervical epithelium. However, the mechanisms that control the expression of this gene in epithelial cells remain unknown. We analyzed the DNA methylation status of the CD86 promoter and a CpG island located in the upstream intergenic region in keratinocyte-derived cell lines. In those cell lines where CD86 is expressed, a high degree of methylation in the CpG island was observed. However, a CpG dinucleotide within the cAMP response element (CRE) in the promoter region was consistently unmethylated and associated to the transcription factor CREB, as demonstrated by ChIP assays. The opposite methylation pattern was observed in cell lines where CD86 is not expressed, affecting also the binding of CREB. The analysis of the DNA methylation pattern of this gene in cells from the spinous and basal layers of normal cervical epithelium showed a similar profile to that observed in cell lines with and without expression of CD86 respectively. Our results indicate that the methylation pattern in the CD86 promoter and CpG island is closely related to the expression of this co-stimulatory molecule in keratinocytes. (C) 2013 Elsevier Inc. All rights reserved.