(A) CD8+/+ and CD8−/− mice were immunized with 100 μg of MOG35–55 and disease course evaluated for 30 days. Representative of three independent experiments (n=15 per condition).
(B) Draining lymph node cells from WT or CD8−/− MOG35–55-immunized mice were CFSE stained and cultured for 5 days with cognate antigen. CFSE dilution in TCRβ+CD8−CD4+ T-cells was evaluated using flow cytometry. Representative histogram (left panels) and cumulative data (right panel) of three independent experiments are shown (n=9 per condition). Δ PF represents difference between proliferation in the presence of antigen and background.
(C) Five-day CFSE cultures were stimulated with PMA/ionomycin/brefeldin-A and stained for surface and intracellular markers. Representative histograms (left panels) and cumulative graphs (right panels) of gated CFSE-low (proliferating) TCRβ+CD8−CD4+ T-cells are shown for the indicated functional molecule (n=10 per condition).
(D) OVA- or MOG-specific CD8 T-cells were transferred into CD8−/− B6 mice i.v. One day later, all mice where immunized with MOG35–55/CFA and EAE clinical disease evaluated for 30 days. Representative of two independent experiments (n=11 per condition).
(E) Flow cytometry data of MOG35–55-responding CD4 T-cells from CD8−/− recipients of MOG-specific or OVA-specific CD8 T-cells. Representative of two independent experiments (n=11 per condition)