The neuropeptide alpha-melanocyte stimulating hormone (alpha-MSH) is recognized as a potent mediator of immune and inflammatory reactions. Accordingly, alpha-MSH in vitro, as well as in vivo, antagonizes the proinflammatory activities of cytokines such as interleukin-1 (IL-1), IL-6, and tumor necrosis factor alpha (TNF alpha). Since the molecular basis of these antiinflammatory effects is not well known, the influence of alpha-MSH on IL-1 beta-induced chemokine production and transcription factor activation was investigated in human keratinocytes. alpha-MSH, in a dose-dependent manner, after 48 h, significantly reduced the IL-1 beta mediated secretion of the C-X-C chemokines IL-8 and Gro alpha. This was confirmed by semiquantitative RT-PCR, which revealed a marked down-regulation in IL-8 and Gro alpha mRNA expression. Furthermore, we determined the effect of alpha-MSH on the IL-1 beta-induced activation of the nuclear factor kappa B (NF kappa B)--a major transcription factor for chemokine genes. Electrophoretic mobility-shift-assays showed that alpha-MSH, in a dose range from 10(-6) to 10(-12) M, significantly downregulated the IL-1 beta-induced activation of NF kappa B 10 minutes after stimulation. Therefore, NF kappa B inactivation by alpha-MSH appears to be a crucial event, one that is responsible for the downregulation of cytokine gene transcription.