Figure 4. The JAK1/2 Inhibitor AZD1480 Suppresses Classical EAE.

(A). C57BL/6 mice were immunized with MOG_35-55 peptide (200 μg) emulsified in CFA containing M. tuberculosis. The mice received i.p. injections of 250 ng of pertussis toxin on days 0 and 2. Vehicle Control (0.1% DMSO) (n=14) or AZD1480 (25 mg/kg) (n=15) was administrated i.p. daily for 10 days starting at the onset of EAE (day 9). Mean ± S.D. of classical EAE clinical scores. (B). Protein extracts from the spinal cord of Vehicle Control or AZD1480 treated mice at day 16 were immunoblotted with the indicated antibodies. (C). CNS-infiltrating mononuclear cells were isolated from the spinal cord of Vehicle Control or AZD1480 treated mice at day 16. Cells were stained with trypan blue and counted. Cells were stained with Abs to CD4, CD11b, Gr-1, CD45, CD11c, Ly-6C and B220, and the percentage of CD11b⁺/Gr-1⁺ neutrophils, CD11b⁺/Ly-6C^hi and CD11b⁺/Ly-6C^lo monocytes, CD11b⁺/CD45^hi macrophages, CD11b⁺/CD45^int microglia, CD11c⁺ dendritic cells, CD4⁺ T-cells and B220⁺ B-cells were gated. The absolute number of cells is shown. (D). Mice were perfused, spinal cord removed, and mRNA from spinal cord of Vehicle Control or AZD1480 treated mice at day 16 was analyzed by qRT-PCR. (E). Sections from the spinal cord of Vehicle Control or AZD1480 treated mice at day 16 were stained with H&E for inflammatory infiltrates and luxol fast blue (LFB) for demyelination. (F). C57BL/6 mice were immunized with MOG_35-55 peptide (200 μg) emulsified in CFA containing M. tuberculosis. The mice received i.p. injections of 250 ng of pertussis toxin on days 0 and 2. Vehicle Control (0.1% DMSO) (n=12) or AZD1480 (25 mg/kg) (n=13) was administrated i.p. after mice reached a clinical score of ∼1.5 for up to 12 days. *p<0.05 and **p<0.001.