FIGURE 6.

VentX regulates maturation of lamina propria DCs. A, LPDCs were isolated from inflamed and control non-inflamed intestinal mucosa of five IBD patients. VentX expression level was determined by real-time PCR. B, LPDCs were transfected with siRNA targeting VentX or control GFP through electroporation. Two days post-transfection, cells were stimulated with LPS for overnight to induce maturation and harvested for flow cytometry analysis of surface markers and intracellular cytokines/chemokines. Filled histograms indicate siGFP transfection. C, bar graph showed the mean fluorescence intensity (MFI) of four independent experiments in B. Results are expressed as the percentage of MFI normalized to control. There was a significant difference between these two groups (p < 0.05) except for CD83, which was not significant (N.S.). D, LPDCs transfected with siGFP or siVentX were analyzed for the mRNA expression levels of indicated genes. There was a significant difference between the two groups (p < 0.05) except for CCL19 and CXCL1. E, knockdown of VentX impaired the migration capability of DCs. F, knockdown of VentX impaired DCs capability to stimulate allogenic T cells proliferation in a mixed lymphocyte reaction. G, DCs were treated with prednisolone (10 μg/ml) for 48 h or mock treated; then VentX level was determined with real time PCR. H, DCs were transfected with GFP or VentX and then treated with PDL as indicated. LPS was added 24 h later, and cells were harvested 48 h after transfection to analyze the mRNA levels of CCL5, IL12A, and TNFα. Results shown are mean ± S.D. of triplicates from one of two independent experiments. I, schematic of VentX regulation of monocyte (Mo)-derived DC differentiation.