BACKGROUND: It is increasingly evident that there is a close connection between the generation of cutaneous inflammatory cytokines and elevated neuropeptide signaling in complex regional pain syndrome (CRPS) patients. Previously, we observed in the rat tibia fracture model of CRPS that activation of caspase-1 containing NALP1 inflammasomes was required for interleukin (IL)-1β production in keratinocytes, and that administration of an IL-1 receptor antagonist (anakinra) reduced the fracture-induced hindpaw mechanical allodynia. We therefore hypothesized that neuropeptides lead to nociceptive sensitization through activation of the skin's innate immune system by enhancing inflammasome expression and caspase-1 activity. METHODS: We determined whether the neuropeptides substance P (SP) and calcitonin gene-related peptide (CGRP) require IL-1β to support nociceptive sensitization when injected into mouse hindpaw skin by testing mechanical allodynia. We then investigated whether these neuropeptides could stimulate production of IL-1β in a keratinocyte cell line (REKs), and could increase the expression of inflammasome component proteins including NALP1 and caspase-1. Finally, we determined whether neuropeptide-stimulated IL-1β production required activation of caspase-1 and cathepsin B. RESULTS: Intraplantar injections of SP and CGRP lead to allodynia in mouse hindpaws but CGRP was approximately 10-fold less potent in causing this response. Moreover, systemic administration of the IL-1 receptor (IL-1R) antagonist anakinra prevented sensitization after neuropeptide injection. Also, mouse skin keratinocytes express IL-1R, which is up-regulated after local neuropeptide application. In vitro data demonstrated that both SP and CGRP increased IL-1β gene and protein expression in REKs in a dose-dependent manner. Furthermore, SP time- and dose-dependently up-regulated NALP1 and caspase-1 mRNA and protein levels in REKs. In contrast, CGRP time- and dose-dependently enhanced NALP1 and caspase-1 mRNA levels without causing a significant change in NALP1 or caspase-1 protein expression in REKs. Inhibition of caspase-1 activity using the selective inhibitor Ac-YVAD-CHO reduced SP and, less effectively, CGRP induced increases in IL-1β production in REK cells. The selective cathepsin B inhibitor CA-74Me inhibited neuropeptide induced IL-1β production in REKs as well. CONCLUSIONS: Collectively, these results demonstrate that neuropeptides induce nociceptive sensitization by enhancing IL-1 β production in keratinocytes. Neuropeptides rely on both caspase-1 and cathepsin B for this enhanced production. Neurocutaneous signaling involving neuropeptide activation of the innate immunity may contribute to pain in CRPS patients.