Fig 2: Plasmodium‐specific memory CD4+ T cells induced under IL‐27 neutralization exhibit enhanced cytokine productionCytokine production was analyzed for PbT‐II cells transferred to B6 mice infected with Pcc and were treated with control (IgG, blue) or anti‐IL‐27 mAb (α‐IL‐27, red) between −1 and 7 days after infection. A, BSplenocytes from IgG and α‐IL‐27‐treated mice collected 7 and 28 days after infection were stimulated with PMA and ionomycin for 4 h (A), and splenic CD4+ T cells were cultured with PbT‐II peptide‐pulsed dendritic cells for 4 h (B), stained for surface markers, fixed, permeabilized, and stained for IFN‐γ, TNF, IL‐10, and IL‐2. Representative flow cytometry profiles (left) of cytokine expression of PbT‐II cells (CD45.1+CD45.2−TCRβ+CD4+) and frequencies of cytokine‐producing PbT‐II cells (right) are shown for control (blue) and anti‐IL‐27 mAb (red)‐treated mice at day 7 (n = 4 mice/group) and day 28 (n = 5 mice/group) days pi. Representative data of two independent experiments are shown.CProportions of IFN‐γ‐ or TNF‐producing cells in response to PbT‐II peptide within indicated subpopulations of control (n = 4) and anti‐IL‐27 mAb‐treated (n = 3) mice.D, ESplenic cells were prepared 28 days after Pcc infection and stained for TCRβ, CD4, CD45.1, CD127, and KLRG1, fixed, permeabilized, and stained for Ki67 to analyze expression in PbT‐II cell subpopulations.DRepresentative histogram plots (left) and summary graphs (right) of Ki67 expression in PbT‐II cells, and isotype control of IgG group (gray) at 28 days pi (n = 4 mice/each group). Data are representative of three independent experiments.ERepresentative histograms (left) and summary graph (right) of median fluorescence intensity (MFI) levels of Ki67 expression of total PbT‐II cells from IgG, and CD127+KLRG1− (orange), CD127−KLRG1+ (red), CD127−KLRG1− (green) PbT‐II subpopulations from anti‐IL‐27 mAb‐treated mice (n = 3 mice/each group). Data are representative of three independent experiments. Data information: Statistical significance was assessed by Student's t test for comparing IgG and anti‐IL‐27 mAb‐treated mouse groups in (A), (B), and (D), and per subpopulation in (C), and one‐way ANOVA followed by Tukey's multiple comparison test in (E). P values (< 0.05) are shown. Error bars represent SD in all graphs. Source data are available online for this figure.