Figure 2.

IL-12 induced the differentiation of polyfunctional CD4⁺ (T)cells. Naive CD4⁺ T cells were stimulated for 5 d with anti-CD3 and anti-CD28 mAbs in the presence of IL-12. The cells were rested and re-stimulated for 6 h with PMA and ionomycin in the presence of BFA. The expression of IL-21, IFN-γ, TNF-α and IL-2 was detected by FACS. Most of IL-21-expressing CD4⁺ T cells co-expressed Th1 cytokine IFN-γ, TNF-α or IL-2. The representative dot plots were shown (A). Gated on IL-21⁻ and IL-21⁺CD4⁺ T cells or IFN-γ⁻ and IFN-γ⁺CD4⁺ T cells, the expression of IFN-γ or IL-21 was analyzed. The representative histogram graphs and statistical data of mean MFI were shown (B). Gated on IL-21⁻IFN-γ⁻, IL-21⁺IFN-γ⁺, IL-21⁺IFN-γ⁻ and IL-21⁻IFN-γ⁺ CD4⁺ T cells, the expression of TNF-α and IL-2 in the 4 different subsets were analyzed. The representative dot plots (C) and statistical data of mean percentages (D) of TNF-α⁻IL-2⁻, TNF-α⁻IL-2⁺, TNF-α⁺IL-2⁻ and TNF-α⁺IL-2⁺ were shown.