Churchill L, Friedman B, Schleimer R P, Proud D
Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, Maryland.
Immunology. 1992 Jan;75(1):189-95.
We have evaluated the capacity of cultured human respiratory epithelial cells (HTE) to produce granulocyte-macrophage colony-stimulating factor (GM-CSF) and have examined the ability of proinflammatory stimuli and of glucocorticoids to modulate the production of GM-CSF by these cells. Conditioned medium (CM) was obtained after 24-hr culture of HTE in the presence or absence of serum (5%) and was assayed for GM-CSF activity using the M-07e cell line, which proliferates in response to GM-CSF and interleukin-3 (IL-3). HTE produced 1.1 +/- 0.7 and 2.1 +/- 1.5 ng GM-CSF/10(6) cells in the presence or absence of serum, respectively (n = 4). The identity of this activity as GM-CSF was established by neutralization with specific antibody to GM-CSF, while antibody to IL-3 was without effect. Dexamethasone (10(-6) M) inhibited basal GM-CSF release to below the limit of detection of the assay (0.12 ng GM-CSF/ml conditioned media). GM-CSF release was significantly enhanced by 10(-6) M histamine (1.6 +/- 0.7 versus 2.13 +/- 0.8 ng GM-CSF/10(6) cells; n = 9) and 5 ng/ml interleukin-1 (IL-1) (0.6 +/- 0.2 versus 3.2 +/- 0.5 ng GM-CSF/10(6) cells; n = 3). Stimulation of GM-CSF release by IL-1 was dose-dependent. A significant increase in GM-CSF activity was observed with 0.1 ng/ml, while maximal stimulation occurred at 5 ng/ml IL-1. In kinetic studies, GM-CSF activity was first detected in CM from cells incubated in the absence of stimulus at 8 hr of incubation, and continued to increase up to 24 hr. IL-1 stimulated GM-CSF activity was detected in CM as early as 4 hr and continued to increase significantly up to 24 hr. Thus, human tracheal epithelial cells release GM-CSF and this release is regulated by inflammatory mediators and glucocorticoids.
我们评估了培养的人呼吸道上皮细胞（HTE）产生粒细胞-巨噬细胞集落刺激因子（GM-CSF）的能力，并研究了促炎刺激物和糖皮质激素调节这些细胞产生GM-CSF的能力。在有或无血清（5%）存在的情况下，将HTE培养24小时后获得条件培养基（CM），并使用M-07e细胞系检测GM-CSF活性，该细胞系会因GM-CSF和白细胞介素-3（IL-3）而增殖。在有或无血清的情况下，HTE分别产生1.1±0.7和2.1±1.5 ng GM-CSF/10⁶个细胞（n = 4）。通过用GM-CSF特异性抗体中和确定该活性为GM-CSF，而IL-3抗体则无作用。地塞米松（10⁻⁶ M）将基础GM-CSF释放抑制至检测限（0.12 ng GM-CSF/毫升条件培养基）以下。10⁻⁶ M组胺（1.6±0.7对2.13±0.8 ng GM-CSF/10⁶个细胞；n = 9）和5 ng/ml白细胞介素-1（IL-1）（0.6±0.2对3.2±0.5 ng GM-CSF/10⁶个细胞；n = 3）可显著增强GM-CSF释放。IL-1对GM-CSF释放的刺激呈剂量依赖性。在0.1 ng/ml时观察到GM-CSF活性显著增加，而在5 ng/ml IL-1时出现最大刺激。在动力学研究中，在无刺激条件下培养的细胞的CM中，在培养8小时时首次检测到GM-CSF活性，并持续增加直至24小时。IL-1刺激的GM-CSF活性最早在4小时时在CM中检测到，并持续显著增加直至24小时。因此，人气管上皮细胞释放GM-CSF，且这种释放受炎症介质和糖皮质激素调节。
