Fig. 4

Inflammation alters monocyte differentiation program in the small intestine. a Small intestine were surgically manipulated to induce inflammation in the small intestine and 3 days after manipulation, intestinal Ly6G⁻SiglecF⁻CD11b⁺CD11c^lo/intCD64⁺ leukocytes (MNP^CD64+ cells) were divided into five subpopulations based on expression of Ly6C, MHCII, and CX3CR1/GFP; namely Ly6C⁺MHCII⁻, Ly6C⁺MHCII^int, Ly6C⁻MHCII^int, Ly6C⁻MHCII^hiCX3CR1^int and Ly6C⁻MHCII^hiCX3CR1^hi. b Representative histograms demonstrating expression of CX3CR1, CD64, CD11c, and CD206 on subpopulations shown in a and neutrophils (CD11b⁺Ly6G⁺) for comparison. c Schematic illustration of the experimental setup for d–g. Purified monocytes were adoptively transferred to recipient mice that underwent surgical manipulation of the small intestine one day before the transfer. Donor cells were recovered from small intestinal lamina propria (siLP) 1, 2 and 5 days after transfer. d Transferred MNP^CD64+ cells recovered from siLP of the manipulated recipients were identified by congenic markers followed by gating as described in left panel of a. e Representative FACS plots demonstrating gradual phenotypic changes of donor MNP^CD64+ cells over time, with respect to their expression of Ly6C and MHCII. f Frequency of Ly6C- and MHCII-defined subsets among donor MNP^CD64+ cells in the intestine at the indicated time points. Data were pooled from at least three independent experiments with a total of 5–8 mice. g Expression of CX3CR1/GFP, CD64, and CD11c on donor MNP^CD64+ cells at day 1, 2, and 5 post-transfer compared to endogenous neutrophils. Results in g are representative of at least three independent experiments each. h Heat map showing hierarchal clustering of gene expression profiles of adoptively transferred MNP^CD64+ cells. Donor monocyte-derived cells were recovered from siLP of manipulated recipients at day 1, 2, or 5 after transfer and transcriptomic analysis was performed by nanostring nCounter analysis. Differentially expressed genes were determined by Qlucore Omics explorer with a statistical significance p-value of 0.05. Data were pooled from 2–3 independent experiments with a total of 4–5 mice for each time point