Figure 3
Polyfunctional LLO-specific CD4 Th1 cells in intestinal tissues. Single-cell suspensions isolated from the spleen, MLN, LP, or IEL during the primary (9 d.p.i.), memory (60 d.p.i.), or secondary (60+7 d.p.i.) responses were stimulated with LLO190–201 peptide in the presence of brefeldin A, with BD leukocyte activation cocktail (PMA/Iono), or with brefeldin A alone (unstim). (a) IFNγ and IL-17A expression was determined by intracellular cytokine staining after 5 h at 37 °C and 5% CO2. Representative zebra plots are gated on live CD45+ CD4+ T cells and numbers within quadrants correspond to the percentage of cells within gates. Data are representative of 3 experiments with at least three mice per group. (b) Pie charts display the multifunctional nature of CD4 T cells. Multifunctionality was determined among an IFNγ+ CD4+ parent population. Numbers in pie charts represent the mean percentage of IFNγ+ cells that produced the indicated cytokines. Data depict the mean of three mice per group and are representative of two experiments. (c) Representative zebra plots are presented to demonstrate the multifunctional CD4 T-cell response. Numbers within gates represent the mean percentage of concatenated cells that produced the indicated cytokines and are representative of two experiments.
