Monocyte-derived dendritic cells (Mo-DC) express functional P2X₇ receptors; however, the expression of these receptors on tissue-derived dendritic cells including epidermal Langerhans cells (LC) is unknown. Using immunolabeling and flow cytometry, we demonstrated that P2X₇ was present on both human epidermal LC and monocyte-derived LC (Mo-LC), as well as on human keratinocytes. The ecto-ATPDase (CD39) was also present on LC, but not keratinocytes. The P2X₇ agonists, 2′- and 3′-0(4-benzoylbenzoyl) adenosine 5′-triphosphate (BzATP) or ATP, but neither adenosine 5′-diphosphate (ADP) nor uridine 5′-triphosphate (UTP), induced ethidium⁺ uptake into these cells. Furthermore, ATP-induced ethidium⁺ uptake into epidermal LC, Mo-LC and keratinocytes was inhibited by the specific P2X₇ antagonist, KN-62 (1-[N,O-bis(5-isoquinolinesulfonyl)-N-methyl-L-tyrosyl]-4-phenylpiperazine). ATP-induced ethidium⁺ uptake into Mo-LC and Mo-DC was 2- and 3-fold greater, respectively, than that for fresh monocytes. P2X₇ activation on LC induced downstream signaling events, as BzATP or ATP, but neither ADP nor UTP, induced shedding of the low-affinity receptor for IgE (CD23) from Mo-LC. This process was inhibited by KN-62. Finally, ATP-induced ethidium⁺ uptake and CD23 shedding were impaired in Mo-LC obtained from subjects homozygous for the loss-of-function Glu-496 to Ala polymorphism in the P2X₇ receptor. These results demonstrate that human LC express functional P2X₇ receptors, and suggest a role for this receptor in the skin immune system.