Regulation of Intestinal Permeability and Epithelial Cell Tight Junctions by the Ubiquitin-Editing Enzyme TNFAIP3

UC-CMFs.Myeloid dendritic cells (DC) generated from peripheral blood mononuclear cells by IL-4/GM-CSF treatment were used as control APCs.T cell phenotype was determined by immunostaining followed by FACS analysis.CFSE-proliferation assays, real-time RT-PCR were used to evaluate activation of Treg cells co-cultured with CMFs.Results: Co-culturing of allogeneic TH0 cells with both N-and UC-CMFs leads to the upregulation of FoxP3 mRNA expression.FACS analysis demonstrated that N-CMF induced generation of FoxP3+ T cells from TH0 cells at a rate of 10.2±3.6%, comparable to those co-cultured with DCs.N-CMFs induced FoxP3+ T cells were CD25highCD127-, express IL-10 and TGFβ and possess suppressive activity.Co-culture of the TH0 cells with UC-derived CMFs induced FoxP3 expression in cells bearing CD127+CD25low T effector cell phenotype.Since FoxP3 expression in CD127+CD25low T effectors has been associated with anergy, our data suggest that in contrast to N-CMFs, UC-derived CMFs have a reduced capacity to induce an active Treg and may lead to the induction of anergic FoxP3+ CD4+ T effector cells.Conclusions: These results support the notion that in normal colonic mucosa CMFs have an anti-inflammatory role and contribute to tolerance by supporting the Treg cell function.Our data also suggest that disruption in the capacity of UC-CMFs to induce active Treg may contribute to the progression of UC.