Figure 3.

Hypothermia increases interleukin (IL)-22⁺ cells in total peripheral blood mononuclear cells (PBMC). (A) PBMC were kept as unstimulated control or stimulated for 7 h with agonistic anti-CD3 (0.2 µg/ml)/-CD28 (0.02 µg/ml) antibodies at 37 or 33°C. Thereafter, brefeldin A (2 µg/ml) was added for another 4 h, followed by intracellular staining and flow cytometry for IL-22 (A,B) and interferon (IFN)γ (C) detection. Left panel (A–C): depicted are percentages of IL-22⁺ cells in total PBMC (n = 6) (A) and the CD4⁺ PBMC fraction (n = 7) (B) as well as IFNγ⁺ cells in total PBMC (n = 6) (C). Data are shown as means ± SEM. **P < 0.01, ***P < 0.001 compared to unstimulated control at the respective temperature; ^##P < 0.01, ^###P < 0.001; statistical analysis, one-way ANOVA with post hoc Bonferroni correction. Right panel: depicted are representative dot blots displaying IL-22⁺ cells in total PBMC (A) and the CD4⁺ PBMC fraction (B) as well as IFNγ⁺ cells in total PBMC (C).