Figure 5. The reduction in IL‐6 production and the suppression of Th17 differentiation rely on the co‐inhibition of LMP7 and LMP2.

• A, B
  Splenocytes from C57BL/6 mice (A) or human PBMCs (B) were incubated (continuous treatment) with DMSO, ONX 0914 (300 nM), PRN1126 (300 nM), ML604440 (300 nM), LU‐001i (300 nM), PRN1126 + ML604440 (300 nM each), or PRN1126 + LU‐001i (300 nM each) and stimulated with LPS overnight. The IL‐6 concentration in the supernatant was analyzed by ELISA. The highest IL‐6 concentration from each experiment was set to 100%. (A) Data are presented as the mean ± s.e.m. of four independent experiments each measured in triplicates. (B) Data are presented as the mean ± s.e.m. of six different healthy donors. All data were statistically compared to the DMSO‐treated group. ***P < 0.001. One‐way ANOVA.
• C
  The differentiation of CD4⁺ T cells isolated from the spleens of C57BL/6 mice that were exposed (continuous treatment) to DMSO, ONX 0914 (300 nM), PRN1126 (300 nM), ML604440 (300 nM), LU‐001i (300 nM), PRN1126 + ML604440 (300 nM each), or PRN1126 + LU‐001i (300 nM each) and that were stimulated with plate‐bound antibodies to CD3/CD28 in the presence of TGF‐β and IL‐6 and scavenging antibodies neutralizing IL‐4 and IFN‐γ was measured in 3‐day cultures. IL‐17 expression was detected by intracellular cytokine staining after a short restimulation with PMA/ionomycin. IL‐17 expression in activated T cells cultured in the absence of Th17‐polarizing conditions is shown as a comparison (no cocktail). Values reflect the percentage of CD4⁺ cells that are IL‐17A⁺. Data are presented as the means ± s.e.m. of three independent experiments each measured in triplicates. All data were statistically compared to the DMSO‐treated group. ***P < 0.001. One‐way ANOVA.