FIG 6.

Functional polarization of effector CD4 T cells in vaccinated CCR2^−/−mice. Wild-type (WT) or CCR2^−/− mice were vaccinated as described in the legend to Fig. 2. On day 8 after vaccination, lung cells were cultured with NP311 peptide, recombinant IL-2, and brefeldin A for 5 h. The percentages of NP311-specific CD4 T cells that produced IL-17α, IFN-γ, IL-2, TNF-α, and GM-CSF were quantified by intracellular cytokine staining. (A) Percentages of IFN-γ/IL-17α-producing cells among the gated CD4 T cells. (B) Percentages of GM-CSF-producing cells among the gated CD4 T cells. (C) Percentages of IL-2/TNF-α-producing cells among the gated IFN-γ-producing CD4 T cells. Cells cultured without the NP311 peptide (unstimulated) served as a negative control. Data in each graph are the mean ± SEM. Mann-Whitney U test; *, **, and *** indicate significance at P values of <0.05, <0.01, and <0.001, respectively. Each independent experiment had 3 to 5 mice per group. Data represent one of three independent experiments.