Figure 4

Pre-treatment of adalimumab attenuates the effects of plasma from Crohn’s disease (CD) patients on paracellular permeability, OCCLUDIN and ZO-1 expression of 3D Caco-2 cysts. Cysts were exposed at the basal side to plasma (37.5% v/v) from healthy controls (HC) or active CD patients with or without pre-incubation of adalimumab (10 or 20 μg/mL) for 24 hours. (a) Representative images showing the intraluminal accumulation of FITC-D4 (green) in Caco-2 cysts. Bar indicates 50 μm. (b) The mean fluorescence intensity of FITC-D4 measured and expressed as the L/BL ratio of the luminal (L) over the basal (BL) compartment. Representative images (c), and fluorescence intensities (d–e) from confocal microscopy analyses of ZO-1, OCCLUDIN in 3D Caco-2 cysts, Images were captured from the middle cross-section of the cysts. Bar indicates 50 μm. All graphs represent the results of three replicate experiments. Data expressed as means ± SEM with at least 8 cysts per subject (6 subjects per group) or per treatment. *P < 0.05; **P < 0.01; ***P < 0.001 by one-way ANOVA and Tukey’s post-hoc test.