217 Immune Cell Model for Burn Immune Reactions Figure 4. Pre-activated T cells after 3 days of culture in (burn-injured) FSEs. (A) Immunohistochemical CD3 staining of an injured FSE. (B) Number of T cells (CD3+ cells) per FSE after isolation using flow cytometry; dashed line indicates the number of T cells added to the transwell. (C) Percentage of CD3+ (T cells) that are CD4+. Percentage of CD4+ T cells that were (D) CD25+; (E) CD25+CD127¯; (F) CXCR3+; (G) CCR4+CCR6+. Experiments were performed in duplicate using keratinocytes and fibroblasts from 6 different donors and T cells from 5 different donors. Only comparisons between T cells in matrix, in uninjured FSEs and in burn-injured FSEs are shown. Statistically significant differences were calculated using Wilcoxon signed-rank test. Significant differences are indicated by asterisks: *: p < 0.05. T cells increase cytokines levels in (burn-injured) full skin equivalents To study the effect of incorporating T cells in (burn-injured) FSEs, cytokine levels were analyzed in the culture medium at day 3 (Figure 5). The levels in FSEs without T cells and the effect of burn injury on these FSEs were reported in the previous section (Figure 3). The inclusion of T cells in the FSEs significantly increased the levels of IFN-γ, IL-4, IL-6, IL-8, IL-10, IL-12p70, IL-17A, IP-10 and TGF-β1, irrespective of burn injury. However, the levels of IL-10 and IP-10 were reduced after burn injury. Significant 7
RkJQdWJsaXNoZXIy MTk4NDMw