Wong C K, Tsang C M, Ip W K, Lam C W K
Department of Chemical Pathology, The Chinese University of Hong Kong, Prince of Wales Hospital, Shatin, Hong Kong.
Allergy. 2006 Mar;61(3):289-97. doi: 10.1111/j.1398-9995.2006.00972.x.
Mast cells play pivotal roles in IgE-mediated airway inflammation and other mast cell-mediated inflammation by activation and chemoattraction of inflammatory cells.
We investigated the intracellular signaling mechanisms regulating chemokine release from human mast cell line-1 (HMC-1) cells activated by stem cell factor (SCF) or tumor necrosis factor (TNF)-alpha.
Chemokine gene expressions were assessed by reverse transcription-polymerase chain reaction, while the releases of chemokines were determined by flow cytometry or enzyme-linked immunosorbent assay (ELISA). To elucidate the intracellular signal transduction regulating the chemokine expression, phosphorylated-extracellular signal-regulated kinase (ERK), phosphorylated-p38 mitogen-activated protein kinase (MAPK) and nuclear translocated nuclear factor (NF)-kappaB-DNA binding were quantitatively assessed by ELISA.
Either SCF or TNF-alpha could induce release from HMC-1 cells of interleukin (IL)-8, monocyte chemoattractant protein (MCP)-1, regulated upon activation normal T-cell expressed and secreted (RANTES), and I-309, while SCF and TNF-alpha induced release of macrophage inflammatory protein (MIP)-1beta and interferon-gamma-inducible protein-10 (IP-10), respectively. Using various selective inhibitors for signaling molecules, we found that the inductions of IL-8, MCP-1, and I-309 were mediated by either SCF-activated ERK or TNF-alpha-activated p38 MAPK, while the induction of IP-10 by TNF-alpha was mediated by both activated p38 MAPK and NF-kappaB. The induction of RANTES by SCF or TNF-alpha was mediated by ERK and NF-kappaB, respectively, and SCF induced MIP-1beta release was mediated by ERK.
The above results therefore elucidated the different intracellular signaling pathways regulating the release of different chemokines from SCF and TNF-alpha-activated mast cells, thereby shedding light for the immunopathological mechanisms of mast cell-mediated diseases.
肥大细胞通过激活和趋化炎性细胞，在IgE介导的气道炎症及其他肥大细胞介导的炎症中发挥关键作用。
我们研究了调节干细胞因子（SCF）或肿瘤坏死因子（TNF）-α激活的人肥大细胞系-1（HMC-1）细胞趋化因子释放的细胞内信号传导机制。
通过逆转录-聚合酶链反应评估趋化因子基因表达，同时通过流式细胞术或酶联免疫吸附测定（ELISA）测定趋化因子释放。为阐明调节趋化因子表达的细胞内信号转导，通过ELISA定量评估磷酸化细胞外信号调节激酶（ERK）、磷酸化p38丝裂原活化蛋白激酶（MAPK）和核转位核因子（NF）-κB-DNA结合。
SCF或TNF-α均可诱导HMC-1细胞释放白细胞介素（IL）-8、单核细胞趋化蛋白（MCP）-1、活化正常T细胞表达和分泌调节因子（RANTES）以及I-309，而SCF和TNF-α分别诱导巨噬细胞炎性蛋白（MIP）-1β和干扰素-γ诱导蛋白-10（IP-10）释放。使用针对信号分子的各种选择性抑制剂，我们发现IL-8、MCP-1和I-309的诱导由SCF激活的ERK或TNF-α激活的p38 MAPK介导，而TNF-α诱导的IP-10由激活的p38 MAPK和NF-κB共同介导。SCF或TNF-α诱导的RANTES分别由ERK和NF-κB介导，SCF诱导的MIP-1β释放由ERK介导。
上述结果因此阐明了调节SCF和TNF-α激活的肥大细胞释放不同趋化因子的不同细胞内信号通路，从而为肥大细胞介导疾病的免疫病理机制提供了线索。
