Figure 3.

Detection of transplanted WT macrophages in vivo and cytokine profiling. [A and B] Polymerase chain reaction [PCR] was used to detect wild-type macrophages harbouring the Il10rb gene; glycosyltransferase-like domain containing 1 [Gtdc1] was used as a control in [A] peritoneal cavity macrophages and [B] cells isolated from the lamina propria of animals treated with: SV129 IL10rb^-/- IBD animals treated with BMDMs, or clodronate liposomes, or a combination of clodronate liposomes and BMDMs, or clodronate liposomes only. Untreated IBD animals, dilutions of WT gDNA and H₂0 are shown as controls [two individual animals/group are shown]. [C and D] Flow cytometry analysis of CD45.1⁺ expression on donor-derived macrophages 6 days after transplantation into Bl6 Il10rb^-/- IBD animals. Animals were either transplanted with BMDMs only or with a combination of clodronate liposomes and BMDMs [Clodro/BMDMs]. [C] Frequency of CD45.1⁺ donor-derived cells among F480^high/CD11b^high peritoneal cavity macrophages; and [D] Frequency of CD45.1+ donor-derived cells among LP macrophages [mean ± SD]. [E] Secretion of IL1b, IL17a, IL27, and GM-CSF by macrophages derived from the peritoneal lavage fluid of different experimental groups [WT untreated, IBD untreated, and IBD+clodro/BMDMs] after LPS stimulation [data represent two independent experiments, individual values with mean ± SD, n = 4–6 per group, cells isolated after 4–6 weeks of treatment]. Significances are calculated by one-way ANOVA with Dunnett’s multiple comparison test, *p <0.05, **p <0.01, ***p <0.001, ****<0.0001, ns not significant. IBD, inflammatory bowel disease; SD, standard deviation; WT, wild-type; LP, laminapropria; LPS, lipopolysaccharide; BBMDM, bone-marrow derived macrophages; ANOVA, analysis of variance.