Expression of CCR7, CD62L and CX3CR1 on CD27-defined T-helper cell subsets. (A) CCR7 expression of T helper cells from blood (PBMCs; left panel), spleen (middle panel) and mediastinal lymph node (Med. LN; right panel) was investigated by five-colour FCM including a live/dead discrimination dye. CD4+ T helper cells (upper left contour plots; black gates) were gated and analysed for CD8α (y-axis) and CD27 (x-axis) expression (lower left contour plots). Gates were set on CD8α-CD27+ (blue gates), CD8α+CD27+ (green gates) and CD8α+CD27 - (red gates) cell subsets for analyses of CCR7 expression indicated by coloured histograms. Numbers show percentage of gated cell subsets. See main text for description of arrow heads and numbers. Representative data from one individual out of six is shown. Per sample at least 1 × 105 lymphocytes were acquired. (B) CD62L and CX3CR1 mRNA expression of total PBMCs and CD27-sorted T-helper cell subsets ex vivo was examined by use of quantitative RT-PCR. The graphs show normalized 2^ΔΔCt values obtained from cells of three animals (symbols) and geometric means (coloured bars) relative to IS. Significant difference is indicated (* p < 0.05).