Figure 3.

Human Tregs from LAD-1 patients lack suppressive function. A, Proliferation of mouse CD4⁺CD25⁻ T cells alone (black bar), with 1:1 ratio of fresh hTregs (hCD25^hi, gray bar) from healthy donors (D1/D2), or LAD-1 patients (P1/P2). White bar is the hTregs stimulated alone. Upper panel represents post-sort flow cytometric FOXP3 staining on the hTregs. Asterisk (*) represents p < 0.05 for the difference between the black and gray bars in each group. B, Proliferation of mouse CD4⁺CD25⁻ T cells alone, or cultured with varying numbers of pre-activated hTregs (act.hCD25^hi) from healthy donor D1, or LAD-1 patient P1. Upper panel represents FOXP3 staining of 48 h pre-activated hTregs. In these assays, the mouse responders were stimulated with mouse APCs and soluble anti-mCD3 while the fresh hTregs were optimally activated with plate-bound anti-hCD3/CD28. The pre-activated hTregs were not restimulated. C, Proliferation of CD4⁺CD25⁻ or D, CD8⁺CD25⁻ T cells from a normal donor alone (black bar) or with 1:1 ratio of fresh hTregs (gray bar) from healthy donors (D1/D2), or LAD-1 patients (P1/P2). In this assay, the human responders and Tregs were stimulated with human APCs and soluble anti-hCD3. Data are representative of two independent experiments. Asterisk (*) represents p < 0.05 for the differences in suppression between healthy donors and LAD-1 patients.