Fig. 11. The IL-12p40₂ has no effect on pure Treg population as well as pure population of CD4+CD25⁻ T cells whereas NO reduces population of CD4+CD25+ T cells as well as the expression of Foxp3 in pure Tregs.

The pure CD4+CD25+ T cells and CD4+CD25⁻ T cells were isolated as described in ‘Material & Methods’. Following isolation, the cells were cultured in twelve-well plates with appropriate concentrations of plate bound anti-CD3 and anti-CD28 with or without p40₂ (50 ng/ml) or DETA-NONOoate (25 μM). After 24 h, normal control, p40₂- and DETA-NONOate-treated pure CD4+CD25⁺ T cells (A) and CD4+CD25⁻ T cells (C) were incubated with appropriately-diluted PE-conjugated anti-CD4 and FITC-conjugated anti-CD25 Abs as described in ‘Materials and Methods’ followed by FACS analysis. After 24 h of treatment, the mRNA expression of Foxp3 and CD25 were analyzed in CD4+CD25⁺ T cells (B) and CD4+CD25⁻ T cells (D) by semi-quantitative RT-PCR.