Identification of human neutrophil-derived cathepsin G and azurocidin/CAP37 as chemoattractants for mononuclear cells and neutrophils.
Identification of human neutrophil-derived cathepsin G and azurocidin/CAP37 as chemoattractants for mononuclear cells and neutrophils.
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人类嗜中性粒细胞衍生的组织蛋白酶G和Azurocidin/cap37鉴定为单核细胞和中性粒细胞的趋化剂。
DOI：
10.1084/jem.186.5.739
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发表时间：
1997-08-29
期刊：
影响因子：
--
通讯作者：
Oppenheim JJ
中科院分区：
文献类型：
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作者：
Chertov O;Ueda H;Xu LL;Tani K;Murphy WJ;Wang JM;Howard OM;Sayers TJ;Oppenheim JJ
巨噬细胞通常先于中性粒细胞浸润到炎症部位。这表明中性粒细胞可能是单核细胞趋化因子的来源。为了鉴定这些假定的单核细胞引诱剂，我们系统地制备了中性粒细胞颗粒，将其裂解，并通过几种反相色谱程序依次纯化释放的蛋白质。根据 SDS-PAGE 分析，层析级分的单核细胞趋化活性测定产生了与 30 kD 蛋白质相关的活性主峰。该蛋白的 NH2 末端序列显示其与组织蛋白酶 G 相同。人组织蛋白酶 G 的单核细胞趋化活性呈剂量依赖性，最佳浓度为 0.5–1 μg/ml。棋盘分析证明，组织蛋白酶 G 对单核细胞具有趋化性而非趋化性。组织蛋白酶 G 诱导的单核细胞趋化性对百日咳毒素部分敏感，这意味着 G 蛋白偶联受体的参与。组织蛋白酶 G 的酶活性与其单核细胞趋化活性相关，因为 DFP 或 PMSF 灭活的组织蛋白酶 G 不再诱导单核细胞迁移。组织蛋白酶 G 的趋化活性也可以被 α1 抗胰凝乳蛋白酶完全阻断，α1 抗胰凝乳蛋白酶是人血浆中存在的胰凝乳蛋白酶样蛋白酶的特异性抑制剂。此外，组织蛋白酶 G 也是中性粒细胞的有效化学引诱剂和 T 细胞的化学动力学刺激剂。在进行这些体外研究的过程中，我们确定来自人中性粒细胞颗粒的 T 细胞趋化剂 azurocidin/CAP37，在剂量为 0.05 至 5 μg/ml 时，对单核细胞和中性粒细胞具有趋化作用。正如体外趋化活性所预测的那样，向 BALB/c 小鼠皮下注射组织蛋白酶 G 导致单核细胞和中性粒细胞浸润。因此，炎症渗出物从中性粒细胞向单核细胞的转变可以至少部分地通过中性粒细胞颗粒蛋白（例如组织蛋白酶G和azurocidin/CAP37）的细胞外释放来介导。
Macrophage infiltration into inflammatory sites is generally preceded by neutrophils. This suggests neutrophils may be the source of chemotactic factors for monocytes. To identify these putative monocyte attractants, we have systematically prepared neutrophil granules, lysed them, and sequentially purified the released proteins by several reverse phase chromatography procedures. Assays for monocyte chemotactic activity of the chromatography fractions yielded a major peak of activity associated with a protein of 30 kD, according to SDS-PAGE analysis. NH2-terminal sequence of the protein revealed this to be identical to cathepsin G. The monocyte chemotactic activity of human cathepsin G was dose dependent with optimal concentration at 0.5–1 μg/ml. Cathepsin G is chemotactic rather than chemokinetic for monocytes, as demonstrated by checkerboard analysis. Cathepsin G–induced monocyte chemotaxis is partially pertussis toxin sensitive implying the involvement of a G protein–coupled receptor. Enzymatic activity of cathepsin G is associated with its monocyte chemotactic activity, since DFP- or PMSF-inactivated cathepsin G no longer induced monocyte migration. The chemotactic activity of cathepsin G can also be completely blocked by α1 antichymotrypsin, a specific inhibitor of chymotrypsin-like proteinases present in human plasma. In addition, cathepsin G is also a potent chemoattractant for neutrophils and a chemokinetic stimulant for T cells. In the course of pursuing these in vitro studies, we established that the T cell chemoattractant, azurocidin/CAP37 from human neutrophil granules, at doses of 0.05 to 5 μg/ml, was chemotactic for monocytes and neutrophils. As predicted from the in vitro chemotactic activity, subcutaneous injection of cathepsin G into BALB/c mice led to infiltration of both mononuclear cells and neutrophils. Thus, the transition of inflammatory exudate from neutrophil to mononuclear cells can be mediated, at least in part, by extracellular release of neutrophil granule proteins such as cathepsin G and azurocidin/CAP37.