Figure 2

Characterization of the ex vivo CD25⁺CD134⁺ T cell assay. (a) Representative dot plots from healthy donors (HD) showing the staining for CD25⁺CD134⁺ T cells in whole blood cultures in response to either control (left) or phytohemagglutinin (PHA) at 5 μg (right). The percentages of CD25⁺CD134⁺ T cells are shown in upper right quadrants. (b) Frequencies of CD25⁺CD134⁺ T cells in response to PHA in 15 IMID patients after 8 years of treatment with TNF antagonists and in 11 HD are shown. Statistical analysis was performed using the Mann-Whitney test to compare IMID and HD subjects and significant p-values are indicated. (c) Frequencies of CD25⁺CD134⁺ T cells were assessed according to the type of biological agent. Statistical analysis was performed using the Mann-Whitney test to compare subjects treated with Eta and Ada. Horizontal bars represent the median values and horizontal dashed lines indicate the cut-off of positive response corresponding to 0.47% (mean 0.098, SD 0.12) for CD4⁺ T cells, and 0.41% (mean 0.11, SD 0.09) for CD8⁺ T cells. The cut-off for a positive response was calculated as the mean response (%) ± 3SD of negative control.