Fig 1: Calcipotriol exerts strong immunosuppressive effects on CD8+ T cells in the presence of CAFs. PBMCs were cultured alone or co-cultured with CAFs in 2D models. Cultures were treated with either DMSO control (filled circle) or 100 nM calcipotriol (blue filled circle) and PBMCs were stimulated with OKT3 (25 ng/ml) for 5 days. (a) Frequency of proliferating CD8+ and CD4+ T cells in the absence or presence of CAFs as determined by CFSE staining (n = 16). (Right) representative CFSE flow cytometry dot plots. (b) Expression of HLA-DR on CD8+ and CD4+ T cells in the absence or presence of CAFs measured by flow cytometry (n = 16). (Right) representative HLA-DR flow cytometry dot plots. (c) Levels of IFN-? (n = 12), GrzB (n = 9), IL-17 (n = 12) and IL-10 (n = 12) from stimulated PBMCs cultured alone or in the presence of CAFs measured by ELISA. (d) Levels of IL-10 from unstimulated PBMCs cultured alone or in the presence of CAFs as measured by ELISA (n = 12). (e) For some co-cultures CAFs were pre-treated with calcipotriol for 48 h (open circle) before adding them to untreated PBMCs. PBMCs were stimulated with OKT3 (25 ng/ml) for 5 days. PMA/Ionomycin was added on day 5 for 4 h. Frequency of proliferating CD8+ T cells as determined by Ki-67 expression and expression of IFN-? and granzyme B in stimulated CD8+ T cells as measured by flow cytometry. (a–d) Lines between dots indicate paired samples. Wilcoxon matched-pairs singed rank test or Friedman’s test were used to detect statistically significant differences for (a–e), respectively. ns, not significant. *p < 0.05, **p < 0.01, ***p < 0.001.