Cysteinyl-leukotrienes are involved in inflammation and act on at least two G-protein-coupled receptors, CysLT1 and CysLT2. However, the role of the CysLT2 receptor as well as its signaling remain poorly understood. Here we show that leukotriene (LT)C4 induced the production of the chemokine IL-8 in endothelial cells. To further study the signaling cascade involved, HEK293 cells were stably transfected with CysLT2 and used to study the transcriptional regulation of the IL-8 promoter. Stimulation of the cells with increasing concentrations of LTC4 resulted in a time- and concentration-dependent induction of IL-8 transcription and protein synthesis. Use of IL-8 promoter mutants with substitutions in their NF-B, AP-1 or NF-IL-6 binding elements revealed an almost total requirement for NF-B and AP-1 elements, and a lesser requirement for the NF-IL-6 element. Overexpression of dominant-negative IBa prevented the IL-8 transactivation induced by LTC4. LTC4 stimulation induced NF-B and AP-1 DNA binding, which involved the formation of a p50/p65 and a c-Jun/c-Fos complex, respectively. Transfection of the cells with a dominant negative (dn) form of PKCe prevented p65 phosphorylation, whereas dnPKCd prevented AP-1 binding. Moreover, dnPKCd, dnPKCe and dnPKC prevented LTC4-induced IL-8 transcription in response to LTC4. Our data show for the first time that LTC4 can act via the CysLT2 receptor to transcriptionally activate chemokine production through induction of NF-B and AP-1 transcription factors. These findings suggest the potential implication of CysLT2 in the inflammatory response through the modulation of chemokine gene transcription.