Fig. 1: Apoptotic T cells and AE-specific CD8+ T cells accumulate in the CNS of mice with EAE.
A, B C57BL/6 mice (n = 6) were immunized with 100 µg MOG35-55 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 14 days, when all the mice had developed EAE, inguinal lymph nodes (LN) and brains and spinal cords (CNS) were collected, and flow cytometry analysis was performed. A Representative staining of active Caspase 3 in gated live single CD3+ CD4+ T cells (left) and of CD44 versus CD40L in gated apoptotic cells (right). B Cumulative analysis in 6 mice. Bars represent means ± SEM. **P < 0.01, by Mann–Whitney test. C, D Mice were immunized as above (n = 5) and sacrificed between days 28 and 48 after EAE induction. As control, not immunized (CTRL) mice were used. Lymphocytes from spleen (SPL) and central nervous system (CNS) were challenged in vitro with MOG35-55 or with AE-peptide pools and IFN-γ ELISpot was performed. The sum of IFN-γ spots, per 106 cells, against all AE-peptide pools was calculated. C Cumulative analysis of IFN-γ production against MOG35-55 (left) or AE-peptides (right) in MOG35-55-immunized (n = 5) or control (n = 4) mice. Bars represent means ± SEM. *P < 0.05, by Mann–Whitney test. D Spearman correlation between IFN-γ response to the indicated peptides and the cumulative disease score. ns, not significant. The data are from a single experiment representative of three independent experiments.
