Takamizawa A, Koyama S, Sato E, Masubuchi T, Kubo K, Sekiguchi M, Nagai S, Izumi T
First Department of Internal Medicine, Shinshu University School of Medicine, Matsumoto, Japan.
J Immunol. 1999 May 15;162(10):6200-8.
We determined whether human lung fibroblasts might release chemotactic activity for neutrophils (NCA) and monocytes (MCA) in response to bleomycin. The human lung fibroblasts supernatant fluids were evaluated for chemotactic activity by a blind well chamber technique. Human lung fibroblasts released NCA and MCA in a dose- and time-dependent manner in response to bleomycin. Checkerboard analysis of supernatant fluids revealed that both NCA and MCA were chemotactic. Partial characterization revealed that NCA was partly heat labile, trypsin sensitive, and predominantly ethyl acetate extractable. In contrast, MCA was partly trypsin sensitive and ethyl acetate extractable. The release of chemotactic activity was inhibited by lipoxygenase inhibitors and cycloheximide. Molecular sieve column chromatography revealed that both NCA and MCA had multiple chemotactic peaks. NCA was inhibited by leukotriene B4 receptor antagonist and anti-IL-8 and G-CSF Abs. MCA was attenuated by leukotriene B4 receptor antagonist, and monocyte chemoattractant protein-1, GM-CSF, and TGF-beta Abs. Leukotriene B4 receptor antagonist and these Abs inhibited the corresponding m.w. chemotactic activity separated by column chromatography. The concentrations of IL-8, G-CSF, monocyte chemoattractant protein-1, GM-CSF, and TGF-beta in the supernatant fluids significantly increased in response to bleomycin. These data suggest that lung fibroblasts may modulate inflammatory cell recruitment into the lung by releasing NCA and MCA in response to bleomycin.
我们确定了人肺成纤维细胞是否会因博来霉素而释放针对中性粒细胞的趋化活性物质（NCA）和针对单核细胞的趋化活性物质（MCA）。采用盲孔室技术评估人肺成纤维细胞的上清液的趋化活性。人肺成纤维细胞因博来霉素而以剂量和时间依赖性方式释放NCA和MCA。对上清液进行棋盘分析显示NCA和MCA均具有趋化性。部分特性分析表明，NCA部分不耐热、对胰蛋白酶敏感，且主要可被乙酸乙酯提取。相比之下，MCA部分对胰蛋白酶敏感且可被乙酸乙酯提取。趋化活性的释放受到脂氧合酶抑制剂和环己酰亚胺的抑制。分子筛柱层析显示NCA和MCA均有多个趋化峰。NCA受到白三烯B4受体拮抗剂以及抗IL-8和G-CSF抗体的抑制。MCA受到白三烯B4受体拮抗剂以及单核细胞趋化蛋白-1、GM-CSF和TGF-β抗体的减弱作用。白三烯B4受体拮抗剂和这些抗体抑制了通过柱层析分离的相应分子量的趋化活性。上清液中IL-8、G-CSF、单核细胞趋化蛋白-1、GM-CSF和TGF-β的浓度因博来霉素而显著增加。这些数据表明，肺成纤维细胞可能通过因博来霉素而释放NCA和MCA来调节炎症细胞向肺内的募集。
