159 Local Immune Response in Burn Patients Absolute number of NK and B cells increase after burn injury FCM analysis showed that the absolute number of NK cells (CD56+ lymphocytes) was higher in burn tissue from PBW 2-3 (Figure 5A). Relative to total leukocyte numbers, NK cells were significantly reduced in burn tissue from PBW 1 and normalized afterwards (Supplementary Figure 2F). In both healthy skin and burn tissue, the majority of the NK cells was CD16¯ (Figure 5B), which is opposed to the NK cell composition in peripheral blood where approximately 90% of the NK cells are CD16+ [19]. Differences in CD16 expression of the NK cells were not observed between healthy skin and burn tissue or between time points. The absolute number of B cells (CD19+ lymphocytes) were higher in burn tissue from PBW 3 (Figure 5C), while the proportion of B cells within the leukocyte population in burn tissue was similar to that of healthy skin (Supplementary Figure 2G). We identified 4 clusters using FlowSOM analysis of the FCM data: CD56+CD16+ NK cells, CD56+CD16¯ NK cells, CD9+CD56+ B cells and CD9lowCD56¯ B cells (Figure 5D). Clustering analysis showed a clear shift towards more CD9lowCD56¯ B cells in burn tissue but no significant differences were detected over time (Figure 5E). 5
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