Italiani Paola, Mazza Emilia M C, Lucchesi Davide, Cifola Ingrid, Gemelli Claudia, Grande Alexis, Battaglia Cristina, Bicciato Silvio, Boraschi Diana
Institute of Biomedical Technologies, National Research Council, Pisa/Segrate, Italy.
Department of Life Sciences, University of Modena and Reggio Emilia, Modena, Italy.
PLoS One. 2014 Feb 3;9(2):e87680. doi: 10.1371/journal.pone.0087680. eCollection 2014.
Monocytes/macrophages are key players in all phases of physiological and pathological inflammation. To understanding the regulation of macrophage functional differentiation during inflammation, we designed an in vitro model that recapitulates the different phases of the reaction (recruitment, initiation, development, and resolution), based on human primary blood monocytes exposed to sequential changes in microenvironmental conditions. All reaction phases were profiled by transcriptomic microarray analysis. Distinct clusters of genes were identified that are differentially regulated through the different phases of inflammation. The gene sets defined by GSEA analysis revealed that the inflammatory phase was enriched in inflammatory pathways, while the resolution phase comprised pathways related to metabolism and gene rearrangement. By comparing gene clusters differentially expressed in monocytes vs. M1 and vs. M2 macrophages extracted from an in-house created meta-database, it was shown that cells in the model resemble M1 during the inflammatory phase and M2 during resolution. The validation of inflammatory and transcriptional factors by qPCR and ELISA confirmed the transcriptomic profiles in the different phases of inflammation. The accurate description of the development of the human inflammatory reaction provided by this in vitro kinetic model can help in identifying regulatory mechanisms in physiological conditions and during pathological derangements.
单核细胞/巨噬细胞是生理和病理炎症各个阶段的关键参与者。为了了解炎症过程中巨噬细胞功能分化的调控机制，我们设计了一种体外模型，该模型基于暴露于微环境条件连续变化的人原代血液单核细胞，概括了反应的不同阶段（招募、启动、发展和消退）。所有反应阶段均通过转录组微阵列分析进行了分析。鉴定出了在炎症不同阶段受到不同调控的不同基因簇。基因集富集分析（GSEA）定义的基因集显示，炎症阶段富含炎症相关通路，而消退阶段则包含与代谢和基因重排相关的通路。通过比较从内部创建的元数据库中提取的单核细胞与M1和M2巨噬细胞中差异表达的基因簇，结果表明该模型中的细胞在炎症阶段类似于M1，在消退阶段类似于M2。通过qPCR和ELISA对炎症和转录因子进行验证，证实了炎症不同阶段的转录组图谱。这种体外动力学模型对人类炎症反应发展的准确描述有助于确定生理条件下和病理紊乱过程中的调控机制。
