Figure 4

Frequencies of IFN-γ (A), IL-2 (B) and IL-17 (C) producing PLPp specific cells in spinal cords of animals immunized with PLPp in CFA and pertussis toxin after stimulation with PLPp, CpG or combined activation. Spinal cords were isolated on day 12 after immunization, and single cell suspensions were made as described in Materials and Methods. Cells were plated at a density of 100.000 cells per well. Raw data is adjusted to 1.000.000 cells per well for a direct comparison with frequencies of cytokine-producing spleen cells. Each bar represents two independent experiments with pooled spinal cord cultures of totally 7 mice (4 mice in the first experiment, 3 mice in the second experiment) due to the small number of mononuclear cells which can be recovered the CNS. Measurements were conducted in duplicate wells each. p values are given in Section 3.