Murine GMP- and MDP-derived classical monocytes have distinct functions and fates [dataset 1]

Monocytes are ephemeral myeloid immune cells that arise from adult hematopoiesis and circulate in the blood. They comprise two main subsets, in mice defined as classical and non-classical monocytes (CM, NCM). Recent fate mapping and transcriptomic analyses revealed that CM themselves are heterogeneous. Here, we report surface markers that allow segregation of murine GMP- and MDP-derived CM, as well as their functional characterization, including fate definition following adoptive cell transfer. GMP-Mo and MDP- Mo gave equal rise to homeostatic CM progeny, such as blood NCM and gut macrophages; the cells however differentially seeded selected other tissues, including the dura mater and lung. Specifically, GMP-Mo and MDP-Mo gave rise to distinct interstitial lung macrophages, linking CM dichotomy to previously reported pulmonary macrophage heterogeneity. Collectively, we provide evidence for the existence of two functionally distinct CM subsets in the mouse, which differentially contribute to peripheral tissue macrophage populations in homeostasis and following challenge. Bone marrow cells were isolated from femur of a 10 week-old C57BL6 female mouse. The cells were stained with antibody panel containing 174 oligo-conjugated antibodies单核细胞（monocytes）是一类起源于成体造血作用的短命髓系免疫细胞，可在血液中循环流通。单核细胞包含两大主要亚群，在小鼠模型中被分为经典型单核细胞（classical monocytes, CM）与非经典型单核细胞（non-classical monocytes, NCM）。近期的命运图谱分析（fate mapping）与转录组学分析（transcriptomic analyses）研究表明，经典型单核细胞本身亦存在异质性。本研究报道了可用于分选小鼠粒细胞-单核细胞祖细胞（granulocyte-monocyte progenitor, GMP）与单核-树突状细胞祖细胞（monocyte-dendritic cell progenitor, MDP）来源的经典型单核细胞的表面标志物，并对其进行了功能表征，包括过继细胞转移（adoptive cell transfer）后的命运追踪。GMP来源的经典型单核细胞（GMP-Mo）与MDP来源的经典型单核细胞（MDP-Mo）均可分化产生稳态下的经典型单核细胞子代，包括血液中的非经典型单核细胞与肠道巨噬细胞；但二者在部分特定组织的定植模式存在差异，例如硬脑膜与肺组织。具体而言，GMP-Mo与MDP-Mo可分化产生截然不同的肺间质巨噬细胞（interstitial lung macrophages），这一发现将经典型单核细胞的二分特性与此前报道的肺巨噬细胞异质性联系起来。综上，本研究证实小鼠体内存在两类功能迥异的经典型单核细胞亚群，它们在机体稳态及受刺激时，可差异性地补充外周组织的巨噬细胞群体。本研究提取了10周龄雌性C57BL6小鼠股骨中的骨髓细胞，使用包含174种寡核苷酸偶联抗体的抗体组合对细胞进行染色。