Endogenous T cells accumulate in K14E7 skin with an enrichment for CCR6-expressing CD4 T cells.

<p>(A) Ear skin tissue was taken from K14E7 mice in addition to control mice expressing the SIY epitope under the K14 promoter. CD4 and CD8 expression was measured on gated CD45⁺CD3⁺ cells. Data is representative of at least 4 mice/group (B) The numbers of CD4⁺ and CD8⁺ T cells in K14E7 skin or control C57 skin were enumerated per square centimetre of ear skin using flow count beads in flow cytometry. Data represents pooled mice from at least two independent experiments. (C) Gated CD3⁺ CD4⁺ T cells from the lymph node or skin of K14E7 mice were analysed for expression of the chemokine receptors, CCR6 and CCR4. The left hand panels show representative plots of isotype and CCR6 antibody staining for CD4⁺ T cells while the graph summarises chemokine receptor staining representative of 6 mice/group from 3 independent experiments. (D) Both K14E7 mice and C57 mice were analysed for the proliferative marker, Ki67, using intracellular staining of lymphocytes derived from the skin and inguinal lymph nodes (iLN). Naive C57 spleen cells (negative control) or C57 spleen cells treated for 3 days with PMA/Ionomycin (positive control) were also analysed. The K14E7 and C57 data represent 7 mice/group derived from 3 independent experiments while controls are spleen cells from a single mouse in 3 independent experiments. The lower right hand panels are representative plots showing isotype and Ki67 staining in K14E7 or C57 mouse skin.