PDF(Pubmed)
Abstract：
Macrophages exhibit a spectrum of behaviors upon activation and are generally classified as one of two types: inflammatory (M1) or anti-inflammatory (M2). Tracking these phenotypes in living cells can provide insight into immune function, but remains a challenging pursuit. Existing methods are mostly limited to static readouts or difficult to employ for multiplexed imaging in complex 3D environments while maintaining cellular resolution. We aimed to fill this void using bioluminescent technologies. Here we report genetically engineered luciferase reporters for long-term monitoring of macrophage polarization via spectral phasor analysis. M1- and M2- specific promoters were used to drive the expression of bioluminescent enzymes in macrophage cell lines. The readouts were multiplexed and discernable in both 2D and 3D formats with single cell resolution in living samples. Collectively, this work expands the toolbox of methods for monitoring macrophage polarization and provides a blueprint for monitoring other multifaceted networks in heterogeneous environments.
摘要：
巨噬细胞在活化时表现出一系列行为,并且通常被分类为两种类型之一:炎性(M1)或抗炎(M2)。追踪活细胞中的这些表型可以提供对免疫功能的洞察，但仍然是一个具有挑战性的追求。现有方法大多限于静态读出,或者难以在保持细胞分辨率的同时在复杂的3D环境中用于复用成像。我们旨在使用生物发光技术填补这一空白。在这里，我们报告了基因工程荧光素酶报告基因，用于通过光谱相量分析长期监测巨噬细胞极化。M1和M2特异性启动子用于驱动巨噬细胞系中生物发光酶的表达。在活样品中,读数以具有单细胞分辨率的2D和3D格式复用和可辨别的。总的来说，这项工作扩展了监控巨噬细胞极化的方法工具箱，并为在异构环境中监控其他多方面网络提供了蓝图。
