Figure 3.

IL-12p35 treatment suppressed adoptive transfer of experimental autoimmune encephalomyelitis (EAE) by encephalitogenic T cells. EAE was induced in mice by active immunization with MOG_35–55-peptide in complete Freund’s adjuvant, and the mice were treated with p35 or PBS. Spleen cells from the mice on day 17 after EAE induction were restimulated ex vivo with MOG_35–55 for 72 h. Activated cells (1 × 10⁷ cells/mouse) were transferred to naïve syngeneic mice and clinical disease monitored until day 17 days after adoptive transfer of encephalitogenic cells. (A) EAE clinical scores. (B,C) On day 17 after adoptive transfer, cells were isolated from the brain or spinal cord and analyzed by intracellular cytokine staining. Cells were gated on CD4⁺ cells, and numbers in quadrants indicate percentage of cells IFN-γ-, IL-17-, and IL-10-expressing CD4⁺ T cells. (D) Proliferative responses of cells in the spinal cord and brain of C57BL/6J recipient mice 17 days after adoptive transfer were assessed by ³H-thymidine incorporation assay. Five replicate cultures were analyzed, and data presented as mean value of CPM of the five replicate cultures. The data are presented as the mean ± SEM of three determinations. ****p < 0.0001; ***p < 0.001; **p < 0.01; *p < 0.05, significantly different from adoptive transfer of MOG_35–55-stimulated with PBS.