126 Chapter 4 it. Thus, within the lymphocyte population, there was an increase in effector cells and Tregs from week 2 onward that show a mixed pro- and anti-inflammatory phenotype. Figure 5. Supervised analysis of blood lymphocyte subsets after severe burn injury. Flow cytometry results of: (A) CD4¯ T cells (CD3+CD4¯ lymphocytes). (B) CD4+ T cells (CD3+CD4+ lymphocytes). (C) Tregs (CD3+CD4+CD25+CD127¯ lymphocytes). (D) CCR4¯CCR6+ CD4+ (non-Treg) T cells; (E) CCR4+CCR6¯ CD4+ (non-Treg) T cells; (F) CCR4+CCR6+ CD4+ (non-Treg) T cells; (G) CCR4¯CCR6+ Tregs; (H) CCR4+CCR6¯ Tregs; (I) CCR4+CCR6+ Tregs. Number of subjects per time interval is shown on top of the graphs. Cell subset concentrations of burn wound patients and healthy controls (HC) are shown as mean (line and dots) ± standard deviation (colored band). Asterisks indicate significant differences in time within the burn patient group (linear mixed model analysis): *p < 0.05. Significant differences of outcomes in burn patients on PBD 0-3 compared to healthy controls are indicated by × (××p < 0.01). Burn Injury Induces High Levels of Circulating Pro-Inflammatory Immune Mediators To study circulating immune mediators induced by burn injury, we screened a broad panel of 33 cytokines, chemokines and growth factors in plasma of burn wound patients from PBD 0 until 48. To highlight significant changes in burn wound patients, data was transformed to fold changes in relation to the levels detected in healthy controls and presented in volcano plots (Figure 6A-F). Pro-inflammatory cytokines IL-6 and IL-8 were increased at all time intervals. Furthermore, we found an increase in chemokines
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