State Key Laboratory of Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing 100094, People's Republic of China.
Immunopharmacol Immunotoxicol. 2010 Dec;32(4):656-62. doi: 10.3109/08923971003671090. Epub 2010 Mar 10.
Although researchers have recently begun to pay more attention to the immunological characteristics of microvascular endothelial cells (MVECs), there are no reports on whether activation of MVECs by interferon-γ (IFN-γ) exerts any influence on the expressions of IFN-α/β. In the present study, we examined the influence of IFN-γ on the expressions of IFN-α/β in rat intestinal mucous MVECs (RIMMVECs). Different concentrations of IFN-γ were used to stimulate cultured RIMMVECs in vitro, and the cells and cell supernatants were collected at different time intervals. The influence of IFN-γ on the expressions of IFN-α/β in the RIMMVECs was examined at the mRNA and protein levels by real-time quantitative PCR and enzyme-linked immunosorbent assay (ELISA), respectively. The results indicated that IFN-γ was able to activate RIMMVECs, thereby leading to upregulated expressions of IFN-α/β. The real-time quantitative PCR analyses indicated that the IFN-α/β mRNA expression levels in RIMMVECs achieved their peak values after stimulation with IFN-γ at 20 ng/mL for 6 h and were increased by 14.88- and 3.82-fold, respectively, when compared with the levels in negative control cells. The ELISA analyses revealed that the IFN-α/β protein expression levels achieved their peak values after stimulation with IFN-γ at 40 ng/mL. The expression of IFN-α protein achieved its peak value at 12 h, while the expression of IFN-β protein achieved its peak value after 6 h. The present results suggest that the expression and secretion of IFNs may participate in the immunologic barrier function of MVECs.
虽然研究人员最近开始更加关注微血管内皮细胞(MVEC)的免疫学特性,但目前尚无关于干扰素-γ(IFN-γ)是否会激活 MVEC 从而影响 IFN-α/β 的表达的报道。本研究检测了 IFN-γ对大鼠肠黏膜 MVEC(RIMMVEC)中 IFN-α/β 表达的影响。体外培养 RIMMVEC 时,用不同浓度的 IFN-γ 刺激细胞,在不同时间点收集细胞和细胞上清液。通过实时定量 PCR 和酶联免疫吸附试验(ELISA)分别检测 IFN-γ在 mRNA 和蛋白水平对 RIMMVEC 中 IFN-α/β 表达的影响。结果表明,IFN-γ能够激活 RIMMVEC,从而上调 IFN-α/β 的表达。实时定量 PCR 分析表明,IFN-α/β 的 mRNA 表达水平在 20 ng/mL IFN-γ刺激 6 h 后达到峰值,与阴性对照组相比,分别增加了 14.88 倍和 3.82 倍。ELISA 分析表明,IFN-α/β 蛋白表达水平在 40 ng/mL IFN-γ刺激下达到峰值。IFN-α 蛋白的表达在 12 h 时达到峰值,而 IFN-β 蛋白的表达在 6 h 时达到峰值。这些结果提示 IFN 的表达和分泌可能参与 MVEC 的免疫屏障功能。