Figure 4
The Th17 cells migrate towards HNSCC milieu. (A) Migration assay with the ChemoTx Sytem as described in Materials and methods. In the wells of the 96-well plate, 20 × 103 HNSCC cells of the cell line PCI-13 were seeded and on the top of the filter 20 × 103 Th17 cells. After 8 h of incubation, the cells in the well of the 96-well plate were harvested and stained with CD3-FITC, CD4-PerCP, IL-17-APC and analysed with flow cytometry. The first column shows the migration of Th17 cells vs DMEM, the second column shows the migration of Th17 cells vs HNSCC cells (n=25) with P<0.05. The box plots show the median (middle line), 25th and 75th percentiles (box), the extreme values (whiskers, which indicate the 90th and 10th percentile) and the outliers (circles). (B) The migration of Th17 cells towards tumour is CCR6/CCL20 dependent. The Th17 cells were incubated with CCR6 blocking antibody. In the chemotaxis assay, these blocked Th17 cells (column 3) show a reduced migration towards HNSCC compared with the unblocked Th17 cells (column 2) (n=15) with P<0.05.
