Department of Immunology, University of Pittsburgh School of Medicine, Pittsburgh, PA.
Center for Systems Immunology, University of Pittsburgh, Pittsburgh, PA.
J Immunol. 2024 Nov 15;213(10):1508-1515. doi: 10.4049/jimmunol.2400215.
Monocytes and macrophages are central to host defense but also contribute to inflammation-associated pathology. Efforts to manipulate monocyte and macrophage function are limited by our ability to effectively quantify the functional programs of these cells. We identified the gene Fth1, which encodes the ferritin H chain, as highly predictive of alveolar macrophage transcriptomic states during LPS-induced lung inflammation and developed an Fth1-mScarlet reporter mouse. In the steady-state lung, high Fth1-mScarlet expression is restricted to alveolar macrophages. In response to LPS-induced lung inflammation, Fth1 reporter activity is robustly increased in monocytes, with its expression reporting genes that are differentially expressed in monocytes versus macrophages. Consistent with this reporter-associated gene profile, within the Lyz2-GFP+CD11b+Ly6C+ gate, the highest Fth1 reporter expression was observed in CD11c+ cells, indicative of monocyte-to-macrophage differentiation. Although Fth1-mScarlet was induced in monocytes responding to either TLR4 ligation or M-CSF-induced macrophage differentiation in vitro, TLR4-dependent expression occurred with greater speed and magnitude. Considering this, we suggest that Fth1-mScarlet expression reports monocyte-to-macrophage differentiation, with increased expression in proinflammatory states. Dissecting macrophage differentiation from inflammatory programs will be enhanced when combining Fth1-mScarlet with other reporter systems. Thus, the Fth1-mScarlet model addresses an important lack of tools to report the diverse spectrum of monocyte and macrophage states in vivo.
单核细胞和巨噬细胞是宿主防御的核心,但也有助于炎症相关的病理。由于我们有效量化这些细胞功能程序的能力有限,因此,我们对单核细胞和巨噬细胞功能的调控作用受到了限制。我们鉴定了编码铁蛋白 H 链的基因 Fth1,该基因在 LPS 诱导的肺部炎症期间高度预测肺泡巨噬细胞转录组状态,并开发了 Fth1-mScarlet 报告小鼠。在肺部稳态中,高 Fth1-mScarlet 表达仅限于肺泡巨噬细胞。在 LPS 诱导的肺部炎症反应中,Fth1 报告基因的活性在单核细胞中显著增加,其表达报告了在单核细胞与巨噬细胞中差异表达的基因。与该报告基因图谱一致,在 Lyz2-GFP+CD11b+Ly6C+门内,观察到 CD11c+细胞中 Fth1 报告基因表达最高,表明单核细胞向巨噬细胞分化。尽管 Fth1-mScarlet 在单核细胞中响应 TLR4 衔接或 M-CSF 诱导的巨噬细胞分化而被诱导,但 TLR4 依赖性表达发生的速度更快,幅度更大。鉴于此,我们认为 Fth1-mScarlet 表达报告单核细胞向巨噬细胞分化,在促炎状态下表达增加。当将 Fth1-mScarlet 与其他报告系统结合使用时,将增强对巨噬细胞分化与炎症程序的区分。因此,Fth1-mScarlet 模型解决了在体内报告单核细胞和巨噬细胞状态多样化的重要工具缺乏的问题。