Research Article

Combined Exposure of Activated Intestinal Epithelial Cells to Nondigestible Oligosaccharides and CpG-ODN Suppresses Th2-Associated CCL22 Release While Enhancing Galectin-9, TGFβ, and Th1 Polarization

Figure 1

Methods for in vitro studies. HT29 cells (IEC) cultured on solid phase plates were preincubated with IFNγ, TNFα, and IL1α (all 10 ng/mL) alone or in combinations for 6 h, washed, and exposed to medium for 24 h (a). IEC cultured on transwell filters (b, c) were basolaterally preincubated with IFNγ and TNFα in the presence or absence of IL1α (all 10 ng/mL) and apically exposed to scGOS/lcFOS±synthetic CpG-ODN for 6 h, washed, and basolaterally exposed to either medium for 24 h (b) or immature DC for 48 h (c), while apically reexposed to medium, scGOS/lcFOS±synthetic CpG-ODN. After 48 h of IEC-DC coculture, ccDC were added to allogeneic naïve T cells for 6 days (MLR) (d) and immune mediators were measured. scGOS/lcFOS: short-chain galacto- and long-chain fructo-oligosaccharides; CpG-ODN: synthetic CpG-ODN type C (TLR9 ligand); imDC: immature DC; ccDC: coculture DC; MLR: mixed lymphocyte reaction.