Wakahara S, Fujii Y, Nakao T, Tsuritani K, Hara T, Saito H, Ra C
Molecular Biology Laboratory, Medicinal Research Laboratoiries, Taisho Pharmaceutical Co., Ltd, Saitama, Japan.
Cytokine. 2001 Nov 21;16(4):143-52. doi: 10.1006/cyto.2001.0958.
Mast cells have been reported to release not only chemical mediators, but also cytokines upon Fc epsilon receptor I(Fc epsilon RI) 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 Fc epsilon RI, and most of them contained tryptase. These pHCMCs sensitized with immunoglobulin E (IgE) and interleukin 4 (IL-4) were activated through cross-linking of Fc epsilon RI. The time-dependent mRNA expression profiles of Fc epsilon RI subunits, cytokines and chemokines in the sensitized pHCMCs upon Fc epsilon RI 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. Fc epsilon RI-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.
据报道，肥大细胞在Fcε受体I（FcεRI）交联时不仅会释放化学介质，还会释放细胞因子。最近，我们建立了一种培养系统，可从外周血单核细胞中获得富含糜蛋白酶的人肥大细胞。然而，这些肥大细胞的功能特性尚未明确。在本研究中，我们检测了外周血来源的人培养肥大细胞（pHCMC）的功能。pHCMC表达功能性FcεRI，并且大多数含有类胰蛋白酶。这些用免疫球蛋白E（IgE）和白细胞介素4（IL-4）致敏的pHCMC通过FcεRI交联而被激活。通过逆转录聚合酶链反应（RT-PCR）检测致敏的pHCMC在FcεRI激活后，FcεRI亚基、细胞因子和趋化因子随时间变化的mRNA表达谱。在活化的pHCMC中检测到了在过敏性炎症中观察到的大多数细胞因子和趋化因子的mRNA。此外，在我们的研究中首次揭示了人肥大细胞中单核细胞趋化蛋白3（MCP-3）以及肥大细胞中肝和活化调节趋化因子（LARC）、胸腺和活化调节趋化因子（TARC）和巨噬细胞衍生趋化因子（MDC）的基因表达。使用酶联免疫吸附测定（ELISA）评估FcεRI介导的细胞因子和趋化因子在蛋白质水平的产生。这些数据表明，能够产生多种细胞因子和趋化因子的pHCMC可能是研究肥大细胞作为过敏性炎症传导者作用的有用候选者。
