Tracing LYVE1+peritoneal fluid macrophages unveils two paths to resident macrophage repopulation with differing reliance on monocytes

SUMMARY Mouse resident peritoneal macrophages, called large cavity macrophages (LCM), arise from embryonic progenitors that proliferate as mature, CD73 + Gata6 + tissue-specialized macrophages. After injury from irradiation or inflammation, monocytes are thought to replenish CD73 + Gata6 + LCMs through a CD73 - LYVE1 + LCM intermediate. Here, we show that CD73 - LYVE1 + LCMs indeed yield Gata6 + CD73 + LCMs through integrin-mediated interactions with mesothelial surfaces. CD73 - LYVE1 + LCM repopulation of the peritoneum was reliant upon and quantitatively proportional to recruited monocytes. Unexpectedly, fate mapping indicated that only ∼10% of Gata6-dependent LCMs that repopulated the peritoneum after injury depended on the LYVE1 + LCM stage. Further supporting nonoverlapping lifecycles of CD73 - LYVE1 + and CD73 + Gata6 + LCMs, in mice bearing a paucity of monocytes, Gata6 + CD73 + LCMs rebounded after ablative irradiation substantially more efficiently than their presumed LYVE1 + or CD73 - LCM upstream precursors. Thus, after inflammatory insult, two temporally parallel pathways, each generating distinct differentiation intermediates with varying dependencies on monocytes, contribute to the replenish hment of Gata6 + resident peritoneal macrophages.