Human SI Mf compartment comprises short- and long-lived subsets that differ in marker expression. (A) The percentage of CCR2⁺ cells among CD14⁺ PBMo (n = 5) and SI Mf subsets (n = 10) was determined by flow cytometry (left). Gates were set according to FMO controls. The expression of CCR2 mRNA was determined by RNaseq analysis; n = 5 (right). (B) The percentage of calprotectin⁺ cells among CD14⁺ PBMo (n = 7) and SI Mf subsets (n = 11) was determined by intracellular staining and flow cytometry. Gates were set according to staining with isotype control antibody. The expression levels of S100A8 and S100A9 mRNA were determined by RNaseq analysis; n = 5 (right). (C) The median fluorescence intensity (MFI) of CD209 relative to MFI of the FMO control on CD14⁺ PBMo (n = 5) and SI Mf subsets (n = 9) was determined by flow cytometry. The expression of CD209 mRNA was determined by RNaseq analysis; n = 5 (right). (A–C, left) Bars represent median values; ns, not significant (t test comparing CD14⁺ PBMo and Mf1). *, P < 0.05; ***, P < 0.001; ****, P < 0.0001 (RM-ANOVA of SI subsets). (A–C, right) Expression values (log2FPKM) were mean-centered by transcript. (D) Representative flow cytometric analysis of recipient-specific HLA-A2 staining on Mf subsets from transplanted duodenum 6 wk after transplantation. Numbers on plots represent recipient⁺ cells (mean ± SD from all patients analyzed at this time point; n = 5). Gray histograms represent staining of HLA-DR⁺ stromal cells from the same sample. (E) The percentage of recipient-derived cells in each Mf subset at 3 (n = 6), 6 (n = 5), and 52 wk (n = 6) after transplantation was determined by flow cytometry. Bars indicate median values. *, P < 0.05; ***, P < 0.001; ****, P < 0.0001 (two-way ANOVA with RM on population).