Figure 2.

Bimekizumab suppresses γδ T cell and Th17 supernatant-mediated osteogenic differentiation of hPDCs. Heat maps summarising inhibition of osteogenic marker expression in hPDCs stimulated with DM or DM supplemented with (A) γδ T cell supernatant or (B) Th17 cell supernatant (± preincubation with 10 µg/mL anti-IL-17A; anti-IL-17F or bimekizumab) for 9 days. Results are expressed as the Log₂ mean fold change in expression compared to growth media control. Quantification of matrix mineralisation after 9 days incubation in (C) γδ T cell supernatant or (D) Th17 cell supernatant (± preincubation with 10 µg/mL anti-IL-17A; anti-IL-17F or bimekizumab). Representative images of matrix mineralisation stained with alizarin red are presented below the quantification graphs. Results are expressed as the mean fold change in expression compared to GM±SEM. ***p<0.001; **p<0.01; *p<0.05; comparisons between each condition by one-way ANOVA with Fisher’s LSD post hoc test (n=3). (E) MSD-multi-array v-plex pro-inflammatory panel expression of IFNγ, TNFα, IL-1β, IL-8, IL-10, IL-17A, IL-17F cytokines from Th17 and γδT cell supernatants. Results are expressed as mean pg/mL ± SEM. ***p<0.001; **p<0.01; comparisons between Th17 and γδT cell supernatants by two-tailed Student’s t-test for each cytokine tested. (F) Expression of DKK1 in hPDCs stimulated with DM or DM supplemented with either γδ T cell or Th17 cell supernatant (± preincubation with 10 µg/mL anti-IL-17A; anti-IL-17F or bimekizumab) for 9 days. Results are expressed as the mean fold change in expression compared to day 0 GM ± SEM. ***p<0.001; **p<0.01; *p<0.05; comparisons between each condition by one-way ANOVA with Fisher’s LSD post hoc test (n=3). ANOVA, analysis of variance; DM, differentiation medium; GM, growth medium; hPDCs, human periosteum-derived cells; IL, interleukin; LSD, least significant difference; MSD, Meso Scale Discovery; Th17, T helper 17; TNF, tumor necrosis factor.