Constitutive secretion of the granule chymase mouse mast cell protease-1 and the chemokine, CCL2, by mucosal mast cell homologues
Constitutive secretion of the granule chymase mouse mast cell protease-1 and the chemokine, CCL2, by mucosal mast cell homologues
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DOI：
10.1046/j.1365-2222.2003.01571.x
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发表时间：
2003-01-01
影响因子：
6.1
通讯作者：
Miller, HRP
中科院分区：
文献类型：
--
作者：
Brown, JK;Knight, PA;Miller, HRP
粘膜肥大细胞（MMC）颗粒特异性β-糜蛋白酶，小鼠肥大细胞蛋白酶-1（mMCP-1），在线虫感染早期，在寄生虫特异性IgE反应形成之前，全身释放到血流中，TGF-β 1通过MMC的同源物诱导mMCP-1的组成性释放。上皮内MMC也可能表达趋化因子CCL 2，目的研究糜酶mMCP-1和趋化因子CCL 2在线虫感染过程中的表达，并比较二者组成性释放的机制。方法通过在TGF-β 1存在下培养骨髓细胞产生MMC同源物，IL-3、IL-9和干细胞因子（SCF）。共聚焦显微镜观察mMCP-1和CCL 2在细胞内的分布。使用高尔基体破坏剂布雷菲德菌素A和放线菌酮阻断蛋白质合成，研究了高尔基体复合体和蛋白质合成在mMCP-1和CCL 2组成型释放中的参与。结果mMCP-1与高尔基体基质蛋白130共定位，但在颗粒中含量最高，而CCL 2在颗粒中未发现，但似乎仅位于高尔基体复合体中。放线菌酮和高尔基体破坏剂布雷菲德菌素A显著抑制mMCP-1的细胞外释放（约40%），表明通过高尔基体复合体的连续蛋白质合成和运输是最佳mMCP-1分泌所需的。结论促进MMC同源物表达和释放mMCP-1的培养条件也促进了CCL 2的表达和释放。组成性释放涉及从头蛋白质合成，并需要一个功能性高尔基复合体，这表明细胞外分泌的机制相似的两个介质。
Background The mucosal mast cell (MMC) granule-specific beta-chymase, mouse mast cell protease-1 (mMCP-1), is released systemically into the bloodstream early in nematode infection before parasite-specific IgE responses develop and TGF-beta(1) induces constitutive release of mMCP-1 by homologues of MMC in vitro . Intraepithelial MMC may also express the chemokine CCL2 (monocyte chemotactic protein-1) during nematode infection but the expression of this chemokine by MMC homologues has not been investigated.Objective To investigate the expression and to compare the mechanisms of constitutive release of the chymase, mMCP-1, and the chemokine, CCL2.Methods MMC homologues were generated by culturing bone marrow cells in the presence of TGF-beta(1) , IL-3, IL-9 and stem cell factor (SCF). The intracellular distribution of mMCP-1 and CCL2 was examined by confocal microscopy. The involvement of the Golgi complex and of protein synthesis in the constitutive release of mMCP-1 and CCL2 was investigated using the Golgi-disrupting agent brefeldin A and cycloheximide to block protein synthesis. Secreted analytes were quantified by ELISA.Results mMCP-1 colocalized with Golgi matrix protein 130 but was most abundant in the granules, whereas CCL2 was not found in the granules but appeared to be located uniquely in the Golgi complex. Extracellular release of mMCP-1 was significantly inhibited (approximate to 40%) by cycloheximide and by the Golgi-disrupting agent brefeldin A, indicating both continuous protein synthesis and transportation via the Golgi complex are required for optimal mMCP-1 secretion. A similar but more marked inhibitory effect with both compounds was demonstrated on the constitutive secretion of CCL2.Conclusion The culture conditions that promote mMCP-1 expression and release by MMC homologues also promote the expression and release of CCL2. Constitutive release involves de novo protein synthesis and requires a functional Golgi complex, suggesting that similar mechanisms of extracellular secretion operate for both mediators.