FIGURE 3.

Cytokine production and reactivity to MBP by CCR2⁺CCR5⁺ T cells in the PB. (A) Memory CD4⁺ T cell subsets purified from PBMC of HS by flow cytometry were stimulated with PMA and ionomycin. Concentrations of IFN-γ and IL-17 in the supernatants were measured by ELISA. Data represent mean ± SD of three HS. (B) Purified CCR2⁺CCR5⁺ T cells (left panel) and CCR2⁻CCR5⁺ T cells (right panel) were stimulated with PMA and ionomycin for 18 h, and the production of IL-17 and IFN-γ was assessed by intracellular cytokine staining. Numbers indicate the frequency (%) of cells in each quadrant. One representative experiment from three independent experiments with PBMC from HS is shown. (C) Purified memory CD4⁺ T cell subsets were cultured in duplicate with irradiated APC in the presence of MBP (10 μg/ml) or OVA (100 μg/ml) for 5 d. Concentrations of IFN-γ and IL-17 in the supernatants were measured by ELISA. Data represent mean ± SD of six MS patients in relapse, three MS patients in remission, and three HS. *p < 0.05, **p < 0.005.