Dror Natalie, Alter-Koltunoff Michal, Azriel Aviva, Amariglio Ninette, Jacob-Hirsch Jasmine, Zeligson Sharon, Morgenstern Avigail, Tamura Tomohiko, Hauser Hansjörg, Rechavi Gideon, Ozato Keiko, Levi Ben-Zion
Department of Biotechnology and Food Engineering, Technion-Israel Institute of Technology, Haifa, Israel.
Mol Immunol. 2007 Jan;44(4):338-46. doi: 10.1016/j.molimm.2006.02.026. Epub 2006 Apr 4.
Interferon regulatory factor 1 (IRF-1) and IRF-8, also known as interferon consensus sequence binding protein (ICSBP), are important regulators of macrophage differentiation and function. These factors exert their activities through the formation of heterocomplexes. As such, they are coactivators of various interferon-inducible genes in macrophages. To gain better insights into the involvement of these two transcription factors in the onset of the innate immune response and to identify their regulatory network in activated macrophages, DNA microarray was employed. Changes in the expression profile were analyzed in peritoneal macrophages from wild type mice and compared to IRF-1 and IRF-8 null mice, before and following 4 h exposure to IFN-gamma and LPS. The expression pattern of 265 genes was significantly changed (up/down) in peritoneal macrophages extracted from wild type mice following treatment with IFN-gamma and LPS, while no changes in the expression levels of these genes were observed in samples of the same cell-type from both IRF-1 and IRF-8 null mice. Among these putative target genes, numerous genes are involved in macrophage activity during inflammation. The expression profile of 10 of them was further examined by quantitative RT-PCR. In addition, the promoter regions of three of the identified genes were analyzed by reporter gene assay for the ability to respond to IRF-1 and IRF-8. Together, our results suggest that both IRF-1 and IRF-8 are involved in the transcriptional regulation of these genes. We therefore suggest a broader role for IRF-1 and IRF-8 in macrophages differentiation and maturation, being important inflammatory mediators.
干扰素调节因子1（IRF-1）和IRF-8，也被称为干扰素共有序列结合蛋白（ICSBP），是巨噬细胞分化和功能的重要调节因子。这些因子通过形成异源复合物发挥其活性。因此，它们是巨噬细胞中各种干扰素诱导基因的共激活因子。为了更好地了解这两种转录因子在先天免疫反应起始中的作用，并确定它们在活化巨噬细胞中的调控网络，我们采用了DNA微阵列技术。分析了野生型小鼠腹腔巨噬细胞在暴露于IFN-γ和LPS 4小时前后的表达谱变化，并与IRF-1和IRF-8基因敲除小鼠进行了比较。在用IFN-γ和LPS处理后，从野生型小鼠提取的腹腔巨噬细胞中265个基因的表达模式发生了显著变化（上调/下调），而在来自IRF-1和IRF-8基因敲除小鼠的相同细胞类型样本中未观察到这些基因表达水平的变化。在这些假定的靶基因中，许多基因参与炎症期间的巨噬细胞活性。通过定量RT-PCR进一步检测了其中10个基因的表达谱。此外，通过报告基因分析对三个已鉴定基因的启动子区域对IRF-1和IRF-8的反应能力进行了分析。总之，我们的结果表明IRF-1和IRF-8都参与了这些基因的转录调控。因此，我们认为IRF-1和IRF-8在巨噬细胞分化和成熟中具有更广泛的作用，是重要的炎症介质。
