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Am J Physiol Lung Cell Mol Physiol 275: L223-L230, 1998;
1040-0605/98 $5.00
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Vol. 275, Issue 2, L223-L230, August 1998

Human lung fibroblasts release chemokinetic activity for monocytes constitutively

Sekiya Koyama1,2, Etsuro Sato1, Tsuyoshi Masubuchi1, Akemi Takamizawa1, Hiroshi Nomura1, Keishi Kubo1, Sonoko Nagai2, and Takateru Izumi2

1 Shinshu University School of Medicine, First Department of Internal Medicine, Matsumoto 390; and 2 Kyoto University Chest Disease Research Institute, Kyoto 606-01, Japan

We determined whether human lung fibroblasts (HLFs) might release mediators that are responsible for monocyte chemokinetic activity (MCA) constitutively. HLF supernatant fluids showed MCA in a time-dependent manner (P < 0.001). Checkerboard analysis of 24- and 72-h supernatant fluids showed that the activity was chemokinetic. Partial characterization of 24- and 72-h supernatant fluids revealed that the mediators released after 24 h were predominantly composed of lipid-soluble activity, and MCA was blocked by lipoxygenase inhibitors. The mediators released after 72 h were predominantly trypsin sensitive and blocked by cycloheximide. Molecular-sieve column chromatography identified four peaks of MCA. A polyclonal antibody to monocyte chemoattractant protein-1 (MCP-1) inhibited MCA by 20% after 24 h and by 40% after 72 h. Granulocyte-macrophage colony-stimulating factor (GM-CSF) and transforming growth factor-beta (TGF-beta ) antibodies attenuated MCA released after 72 h by 30 and 10%, respectively. These antibodies inhibited corresponding molecular-weight peaks separated by molecular-sieve column. The concentrations of MCP-1, GM-CSF, and TGF-beta were 4,698 ± 242, 26.8 ± 3.8, and 550 ± 15 pg/ml, respectively. A leukotriene B4 (LTB4)-receptor antagonist attenuated the total MCA and the lowest molecular weight peak of MCA. The concentrations of LTB4 were 153.4 ± 12.4 (24 h) and 212 ± 16.6 (72 h) pg/ml. These findings suggest that HLFs may modulate the recruitment of monocytes into the lung by releasing MCP-1, GM-CSF, TGF-beta , and LTB4 constitutively.

monocyte chemoattractant protein-1; granulocyte-macrophage colony-stimulating factor; transforming growth factor-beta ; leukotriene B4


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