Single-cell transcriptomics identifies the differentiation trajectory from inflammatory monocytes to pro-resolving...

Both monocytes and macrophages are heterogeneous populations. It was traditionally understood that Ly6C^hi classical (inflammatory) monocytes differentiate into pro-inflammatory Ly6C^hi macrophages. Accumulating evidence has suggested that Ly6C^hi classical monocytes can also differentiate into Ly6C^lo pro-resolving macrophages under certain conditions, while their differentiation trajectory remains to be fully elucidated. The present study with scRNA-seq and flow cytometric analyses reveals that Ly6C^hiPD-L2^lo classical monocytes recruited to the allergic skin lesion sequentially differentiate into Ly6C^loPD-L2^hi pro-resolving macrophages, via intermediate Ly6C^hiPD-L2^hi macrophages but not Ly6C^lo non-classical monocytes, in an IL-4 receptor-dependent manner. Along the differentiation, classical monocyte-derived macrophages display anti-inflammatory signatures followed by metabolic rewiring concordant with their ability to phagocytose apoptotic neutrophils and allergens, therefore contributing to the resolution of inflammation. The failure in the generation of these pro-resolving macrophages drives the IL-1α-mediated cycle of inflammation with abscess-like accumulation of necrotic neutrophils. Thus, we clarify the stepwise differentiation trajectory from Ly6C^hi classical monocytes toward Ly6C^lo pro-resolving macrophages that restrain neutrophilic aggravation of skin allergic inflammation.