Fig. 3: The effects of cortisol on cytokine production and immune cell migration.

a A result of cytokine antibody array. Especially, among cytokines associating with lymphocyte migration, interleukin (IL)-8, IL-6 and chemokine (C–C motif) ligand 5 (CCL5) protein expression appeared to be slightly reduced by exposure to hydrocortisone (HC) [100 nM, 24 h, in RPMI 1640 medium containing 10% foetal bovine serum (FBS)]. b–e The effect of HC on mRNA expression of IL-8, IL-6 and CCL5 in A549 (b), LK2 (c), NCI-H23 (d) and PC3 (e). Data were calculated by fold change versus control (Co). mRNA expression of IL-8 and IL-6 was significantly reduced by HC treatment in four cell lines, and that of CCL5 was significantly reduced in three cell lines (100 nM, 48 h, in RPMI 1640 medium containing 10% FBS for A549 and LK2, in charcoal-stripped RPMI 1640 medium containing 10% charcoal-stripped FBS for NCI-H23 and PC3). Dose dependency was confirmed in A549 and LK2. Glucocorticoid receptor (GR) dependency was confirmed by knockdown of GR using siGR in A549 and LK2. f–i Peripheral blood mononuclear cell (PBMC) migration assay. f The images of the lower chamber using conditioned medium (CM)—control and HC (100 nM) (bar: 200 μm). g Treatment by HC (100 nM, 48 h RPMI 1640 medium containing FBS) inhibited the migration ability of PBMC in LK2. h, i Assessment of the subpopulation of migrated cells using Papanicolaou staining and immunostaining for CD3 and CD8. h Representative images of Papanicolaou staining and immunocytochemistry for CD3 and CD8. i HC treatment did not alter the ratio of the number of CD3- or CD8-positive T cells to the total migrated cells. Values are mean ± standard deviation (n = 3). *p value < 0.05, significant.