Figure 7. CD4⁺Foxp3⁺ T cells in the skin have regulatory function.

Normal skin biopsies were collected from healthy individuals or form discard surgery skin and cells were isolated by enzymatic digestion. (A) Skin cells and PBMC were stained in parallel and analysed by multi parameter flow cytometry. Live cells were gated on either CD4⁺Foxp3⁺ or CD4⁺Foxp3⁻ (left dotplot) and representative expression of CD25, CD127 and CD39 is shown in the histograms. (B) Skin cells were fractionated into CD25⁺ and CD25⁻ fraction using MACS beads. CD25 depleted cells and mixed CD25⁻/CD25⁺ cells (1:2 ratio) were stimulated o/n with antiCD3/antiCD28 and TNF-α production was measured by intracellular cytokine staining. Dotplots are gated on CD4⁺Foxp3⁻ cells to allow a comparison between the two samples.