Hawkins D L, MacKay R J, MacKay S L, Moldawer L L
Department of Large Animal Clinical Sciences, College of Veterinary Medicine, University of Florida, Gainesville 32610, USA.
Vet Immunol Immunopathol. 1998 Nov 6;66(1):1-10. doi: 10.1016/s0165-2427(98)00181-0.
To investigate the ability of recombinant human interleukin 10 (rhuIL-10) to suppress the release of inflammatory mediators from lipopolysaccharide (LPS)-stimulated equine macrophages, rhuIL-10 was added to equine peritoneal macrophage monolayers at concentrations of 0, 0.1, 1, 10, or 100 ng/ml. Thirty minutes later, LPS (E. coli O55:B5) was added at final concentrations of 0, 1, 10, 100 ng/ml. Macrophages were incubated for 16 h at 37 degrees C, then supernates were harvested and assayed for tumor necrosis factor (TNF) activity (L929 cytotoxicity), interleukin-6 (IL-6) activity (B9 proliferation), prostaglandin E2 concentration (ELISA), and nitric oxide (Griess reaction for nitrite). Preincubation of LPS-stimulated peritoneal macrophages with rhuIL-10 caused significant (P<0.05) reduction in secretion of TNF, IL-6, and PGE2, in a dose-dependent manner. Of the inflammatory mediators, TNF was most sensitive to the effects of rhuIL-10. At concentrations of rhuIL-10> or =1 ng/ml, TNF activity in the supernate was inhibited significantly at all concentrations of LPS. At one or more LPS concentrations, there was significant inhibition of each mediator in the presence of 1 ng rhuIL-10/ml and, at 100 ng/ml, rhuIL-10 significantly inhibited production of each mediator at all LPS concentrations tested. When data were expressed as a percentage of control values and pooled across all LPS concentrations, both PGE2 and TNF values were significantly reduced at rhuIL-10 concentrations of > or =1 ng/ml, whereas IL-6 was inhibited significantly at concentrations of > or =10 ng rhuIL-10/ml. Tumor necrosis factor production was more completely suppressed (7.8% of control) by the highest concentration of rhuIL-10(100 ng/ml) than was PGE2 (27.2%) or IL-6 (43.8%). Nitrite was not detected in any supernate from peritoneal macrophage monolayers.
为研究重组人白细胞介素10（rhuIL-10）抑制脂多糖（LPS）刺激的马巨噬细胞释放炎性介质的能力，将rhuIL-10以0、0.1、1、10或100 ng/ml的浓度添加到马腹膜巨噬细胞单层中。30分钟后，以0、1、10、100 ng/ml的终浓度添加LPS（大肠杆菌O55:B5）。巨噬细胞在37℃孵育16小时，然后收集上清液并检测肿瘤坏死因子（TNF）活性（L929细胞毒性）、白细胞介素-6（IL-6）活性（B9增殖）、前列腺素E2浓度（ELISA）和一氧化氮（亚硝酸盐的格里斯反应）。用rhuIL-10预孵育LPS刺激的腹膜巨噬细胞可导致TNF、IL-6和PGE2的分泌呈剂量依赖性显著（P<0.05）降低。在炎性介质中，TNF对rhuIL-10的作用最敏感。当rhuIL-10浓度≥1 ng/ml时，在所有LPS浓度下，上清液中的TNF活性均被显著抑制。在一个或多个LPS浓度下，存在1 ng rhuIL-10/ml时，每种介质均受到显著抑制，在100 ng/ml时，rhuIL-10在所有测试的LPS浓度下均显著抑制每种介质的产生。当数据表示为对照值的百分比并汇总所有LPS浓度时，当rhuIL-10浓度≥1 ng/ml时，PGE2和TNF值均显著降低，而当rhuIL-10浓度≥10 ng/ml时，IL-6受到显著抑制。最高浓度的rhuIL-10（100 ng/ml）比PGE2（27.2%）或IL-6（43.8%）更能完全抑制肿瘤坏死因子的产生（为对照的7.8%）。在腹膜巨噬细胞单层的任何上清液中均未检测到亚硝酸盐。
