Figure 1

Effects of plasma from patients with Crohn’s disease (CD) on paracellular permeability in 3D Caco-2 cysts. (a) Representative Caco-2 cysts with luminal and basal compartments indicated. The bar indicates 100 µm (b) Representative images showing the intraluminal accumulation of FITC-dextran-4 (FITC-D4, green) in the cysts treated with plasma (37.5% v/v) from healthy controls (HC), active or inactive CD patients, or EGTA (2 mM, positive control) for 24 hours. Images were captured from the middle of the cysts using confocal microscopy. The bar indicates 50 µm. (c) The mean fluorescence intensity of FITC-D4, expressed as the ratio of the luminal (L) over the basal (BL) compartment, after the abovementioned plasma exposure. The L/BL ratio of EGTA exposure was set to 1. (d) Effect of the abovementioned plasma treatments on LDH release in Caco-2 cysts. Triton X-100 was used as positive control to induce maximum LDH leakage. Data are reported as percentage of maximum LDH release. All graphs are expressed as means ± SEM of three independent experiments with 6 subjects per group and at least 8 cysts per subject. *P < 0.05; **P < 0.01; ***P < 0.001 by one-way ANOVA and Tukey’s post-hoc test.