Fig. 3.

γδT17 cells expand due to DC-dependent proliferation. a Ex vivo staining of WT and IL-17r^−/− cLN cells for proliferation marker, Ki-67, to calculate total proliferating cells and contribution of γδ T and specifically CCR6⁺CD27⁻ γδ T to proliferation. Representative of two to three experiments. *p < 0.05, ***p < 0.001. b CFSE in vitro assay with no stimulation, LPS stimulation, and IL-7 titrated stimulation of total cLN cells from WT and IL-17r^−/− mice for 5 days then re-stimulated for 5 h with PMA/Ionomycin to examine proliferation of γδT17. Gated on total γδ T cells. Dot plots and histograms representative of five experiments. c Depletion of CD11c⁺ cells from IL-17r^−/− cLNs or adding back of CD11c⁺ cells to examine γδT cell proliferation after 5 days culture. Cells were gated on 7AAD⁻CD3⁺γδTCR⁺ cells. Representative of five experiments. *p < 0.05, **p < 0.01. d Immunohistochemistry staining of WT and IL-17r^−/− cLNs with pan-γδTCR (green), CD11c (red), and DAPI (blue) to visually show increase in γδ T cells and CD11c⁺DCs. Representative of cLNs from three different mice. Scale bar 20 μm. e High magnification (×20) of cLN image to show γδ T cells (red) and CD11c⁺DC (green) interaction in situ in IL-17r^−/− mice (arrow)