GENE EXPRESSION PROFILES FOR FcϵRI, CYTOKINES AND CHEMOKINES UPON FcϵRI ACTIVATION IN HUMAN CULTURED MAST CELLS DERIVED FROM PERIPHERAL BLOOD

Abstract

Mast cells have been reported to release not only chemical mediators, but also cytokines upon Fcϵ receptor I(FcϵRI) cross-linking. Recently, we have established a culture system to derive chymase-rich human mast cells from mononuclear cells in peripheral blood. However, the functional properties of these mast cells have remained unrevealed. In this study, we examined the functions of peripheral blood-derived human cultured mast cells (pHCMCs). pHCMCs expressed functional FcϵRI, and most of them contained tryptase. These pHCMCs sensitized with immunoglobulin E (IgE) and interleukin 4 (IL-4) were activated through cross-linking of FcϵRI. The time-dependent mRNA expression profiles of FcϵRI subunits, cytokines and chemokines in the sensitized pHCMCs upon FcϵRI engagement were examined by reverse transcriptase polymerase chain reaction (RT-PCR). mRNA for most of cytokines and chemokines, which were observed in allergic inflammation, was detected in activated pHCMCs. In addition, gene expression for monocyte chemoattractant protein 3 (MCP-3) in human mast cells, and liver and activation-regulated chemokine (LARC), thymus and activation-regulated chemokine (TARC) and macrophage-derived chemokine (MDC) in mast cells was revealed for the first time in our study. FcϵRI-mediated cytokine and chemokine production at protein level was evaluated using enzyme-linked immunosorbent assay (ELISA). These data suggest that pHCMCs, which are capable of producing a variety of cytokines and chemokines, can be a useful candidate for investigating roles of mast cells as a conductor for allergic inflammation.