Fig. 7

Human effector/memory Th17 cells preferentially migrate across the BSCFB but in a CCR6/CCL20 independent manner. (A) Total amount (pg) of CCL20 secretion towards the basolateral (top compartment) and apical (bottom compartment) side of unstimulated and 16 h pro-inflammatory cytokine (76IU/mL TNFα + 20IU/mL IFNγ) stimulated HIBCPP monolayers as determined by ELISA. Bar graph shows the mean ± SD of three independent experiments measured in triplicates. (B) Percentage of human Th17 cells migrated across laminin coated Millicell® filters towards the lower compartment in the presence or absence of 500 ng/mL human recombinant CCL20. Bar graph shows the mean ± SD of three independent experiments measured in triplicates. (C) Percentage of human Th1, Th1*, Th2 and Th17 cells that migrated from the basolateral to the apical side across 16 h pro-inflammatory cytokine (76IU/mL TNFα + 20IU/mL IFNγ)-stimulated HIBCPP monolayers after CCR6 inhibition or vehicle control treatment. Dot plot shows the mean ± SD of 2 independent experiments done in triplicates with two different human Th17-cell donors. (D) Percentage of human Th17 cells that migrated from the basolateral to the apical side across 16 h pro-inflammatory cytokine (76IU/mL TNFα + 20IU/mL IFNγ)-stimulated HIBCPP monolayers in the presence or absence of additional 500 ng/mL human recombinant CCL20 in the basolateral (top compartment) or apical (bottom compartment) compartment, and in the presence or absence of CCR6 inhibition and vehicle control treatment. Bar graph shows the mean ± SD of three independent experiments. Statistical analysis: (B) one-way ANOVA and (A, C and D) two-way ANOVA (*p < 0.05, **p < 0.01, ***p < 0.001 and ****p < 0.0001)