Fig. 2: AE-specific CD8⁺ T cells display an effector/memory phenotype in the CNS of mice with EAE.

C57BL/6 mice (n = 8) were immunized with 100 µg MOG_35-55 and 400 µg heat-killed M. tuberculosis in IFA at day 0, and received 200 ng PTX i.v. at days 0 and 2. After 28 days, spleens (SPL) and brains and spinal cords (CNS) were collected, and flow cytometry analysis was performed. A Cumulative analysis of CD8⁺ T cell frequency in SPL and CNS of MOG_35-55-immunized or not immunized (CTRL, n = 4) mice. B Representative stainings with pooled dextramers (Dextr) to the 3 selected peptides, VIM_66-74 (SAVRLRSSV), MYH9_120-127 (VINPYKNL), MYH9_761-768 (VFFRAGVL), in gated live single CD8⁺ T cells, from SPL and CNS of mice with EAE (upper panels). CD44 expression in gated dextramer⁺ (red) or dextramer^– (black) CD8⁺ T cells (lower panels). C Cumulative analysis of dextramer⁺ cell percentage in SPL and CNS of MOG_35-55-immunized or not immunized (CTRL, n = 4) mice. D Frequency of CD44⁺ CD62L^– effector/memory cells in total or dextramer⁺ CD8⁺ T cells from SPL and CNS of MOG_35-55-immunized mice. E Frequency of cells producing IFN-γ, TNF or GM-CSF in gated CD44⁺ among total or dextramer⁺ CD8⁺ T cells from SPL and CNS of MOG_35-55-immunized mice. Samples with insufficient numbers of dextramer⁺ events were not included in the analysis. F Spearman correlation between cytokine production in CD44⁺ dextramer⁺ cells and cumulative disease score. The data are from a single experiment representative of two independent experiments. Bars represent means ± SEM. *P < 0.05, **P < 0.01, ***P < 0.005, by Mann-Whitney test. nd, not determined. ns, not significant.