Glycogen Synthase Kinase-3β regulates antigen/SCF-mediated cytokine production and SCF-mediated chemotaxis in human mast cells (93.6)

Abstract

Abstract Glycogen Synthase Kinase-3β (GSK3β) is a serine threonine kinase downstream of PI3K, which has been implicated in cellular responses including tumorigenesis; cell growth, migration and survival; and cytokine generation. In this study we investigated the role of GSK3β in antigen/SCF-mediated mast cell responses. Short hairpin RNA (shRNA), introduced by lentiviral transfection, was used to knockdown GSK3β in human mast cells. Cytokine production and activation of transcription factors were measured by ELISA and immunoblot analysis. SCF-mediated mast cell chemotaxis was assessed with a transwell migration assay. GSK3β knockdown significantly decreased antigen/SCF-mediated GM-CSF and IL-8 production and SCF-mediated mast cell chemotaxis compared to control treated cells. The decrease in chemotaxis and cytokine production was associated with a pronounced reduction in antigen/SCF-mediated phosphorylation of JNK and the transcription factors c-jun and ATF2. These data are consistent with the conclusion that GSK3β is an essential component of the signalling cascade that regulates antigen/SCF-mediated transcriptional activity and chemotaxis in human mast cells. Thus, targeting GSK3β?may provide one means for down-regulation of these mast cell responses when associated with the allergic inflammatory reaction. Supported by the Division of Intramural Research, NIH, USA.