Figure 1.

Gating for flow analysis of T-cell subsets. (a) CD3⁺ and CD4⁺ T cells. Whole blood cells were surface stained with fluorochrome-labeled monoclonal antibodies to CD3, CD4 and CD45. Lymphocytes were gated on the basis of CD45 and granularity (side scatter). CD3⁺ and CD4⁺ T cells were selected on the basis of CD3 and/or CD4 expression. (b) CD4⁺ naive T cells. MNCs were surface stained with fluorochrome-labeled monoclonal antibodies to CD4 and CD45RA. CD4⁺ T cells were gated, and naive CD4⁺ T cells were selected on the basis of CD45RA expression. (c) CD4⁺ Treg. MNCs were stained with fluorochrome-labeled monoclonal antibodies to CD4, CD45RA and FoxP3. Events were gated to the CD4⁺ T cells, and FoxP3⁺ subsets were selected on the basis of CD45RA and FoxP3 expression. CD45RA⁺FoxP3^low, thymus-derived rTreg; CD45RA⁻FoxP3^high, aTreg; CD45RA⁻FoxP3^low, activation-induced FoxP3⁺ T cells.