Identification of Inducible Murine Mast Cell Genes by Suppression PCR - Based Subtractive Hybridization

Ha, Youn Mun

Abstract

The mast cell is an essential effector cell in allergic inflammation through its capacity to respond to IgE dependent activation. Mast cells also participate in the modulation of physiologic processes, but the role of mast cell in these processes is still unclear. Recently, the number of structurally defined chernoattractants for leukocytes has greatly increased, owing to largely to the identification of the chemokine superfamily. In this study we examined the pattern of expression of chemokines and their receptors in HMC-1 after treatment with PMA/A23187 and/or LPS using RT-PCR and ELISA. Messenger RNA of IL-8, the representative CXC chemokine, was induced after PMA/ A23187 treatment. All of the CC chemokines tested, except eotaxin, were induced after PMA/A23187 treatment. CCR1, CXCR2, CXCR3 and CXCR4 were expressed in all test groups regardless of activation. CCR3 was expressed only at 3 hours of activation. CCR2, CCR5 and CXCR1 were not expressed in mast cell line. Production of most of chemokine proteins was not detected in resting state and increased significantly after 3 hours of activation with PMA/A23187. The effect of LPS treatment was negligible. MCP-1 protein was always produced without activation and accurnulated in a time-dependent rnanner. These data suggest that the expression of mRNA and protein of chemokines and chemokine receptors are regulated transcriptionally and translationally. Human mast cell may respond to various stimuli by producing chemokines and their receptors to regulate their function and may act autonomously or through other inflammatory cell that they recruited.