Elevated IL-4 and GATA-3 and committed Th2 cells in T-bet−/− mice. (A) Naive CD4+ T cells and whole CD4+ T cells from T-bet−/−, T-bet+/+, and wild-type mice were stimulated with anti-CD3/anti-CD28 and maintained under Th1 conditions (IL-12 only) and expanded with IL-2. On day 6, the cells were subjected to intracellular cytokine staining after restimulation with PMA/ionomycin. These data are representative of results obtained from three independent experiments. (B) Naive CD4+ T cells from T-bet−/− and wild-type mice were stimulated with anti-CD3/anti-CD28 under Th1 conditions (IL-12 only) or neutral (no cytokines or antibodies added) conditions for 48 h, and then culture supernatants were harvested for IFN-γ and IL-4 determination by ELISA. The data shown are averages ± SE derived from three independent experiments. (C) Naive CD4+ T cells from T-bet−/− and wild-type mice were stimulated with anti-CD3/anti-CD28 under Th1 conditions (IL-12 only) or neutral conditions and maintained for the time periods shown in the figure. At each time point, cells were harvested and total RNAs were isolated and subjected to real-time PCR analysis for GATA-3 mRNA quantification. Independently, total RNAs from freshly isolated naive and memory CD4+ T cells were prepared for day 9 samples. The data shown are averages ± SE derived from three independent experiments.
Elevated IL-4 and GATA-3 and committed Th2 cells in T-bet−/− mice. (A) Naive CD4+ T cells and whole CD4+ T cells from T-bet−/−, T-bet+/+, and wild-type mice were stimulated with anti-CD3/anti-CD28 and maintained under Th1 conditions (IL-12 only) and expanded with IL-2. On day 6, the cells were subjected to intracellular cytokine staining after restimulation with PMA/ionomycin. These data are representative of results obtained from three independent experiments. (B) Naive CD4+ T cells from T-bet−/− and wild-type mice were stimulated with anti-CD3/anti-CD28 under Th1 conditions (IL-12 only) or neutral (no cytokines or antibodies added) conditions for 48 h, and then culture supernatants were harvested for IFN-γ and IL-4 determination by ELISA. The data shown are averages ± SE derived from three independent experiments. (C) Naive CD4+ T cells from T-bet−/− and wild-type mice were stimulated with anti-CD3/anti-CD28 under Th1 conditions (IL-12 only) or neutral conditions and maintained for the time periods shown in the figure. At each time point, cells were harvested and total RNAs were isolated and subjected to real-time PCR analysis for GATA-3 mRNA quantification. Independently, total RNAs from freshly isolated naive and memory CD4+ T cells were prepared for day 9 samples. The data shown are averages ± SE derived from three independent experiments.
