FIGURE 1.
Temporal IL‐22 overexpression via hydrodynamic gene delivery ameliorates experimental autoimmune encephalomyelitis (EAE) pathology and disease scores. (a) IL‐22 plasmid or empty plasmid was injected on day −2, and EAE was induced by MOG35‐55 immunization on day 0. Mice were monitored for 3–4 weeks, and EAE disease was scored. (b) Spinal cords were removed and stained with Luxol fast blue to quantify demyelination 3 weeks after EAE induction (n = 11 mice for both groups). (c) A representative picture of Luxol fast blue staining. (d) Spinal cords and brain tissues from IL‐22 plasmid‐ or empty plasmid‐injected mouse groups were harvested 3 weeks after EAE induction, and infiltrating lymphocytes were quantified based on IFN‐γ, IL‐17A and GM‐CSF production. Absolute number or percentages of indicated cytokine‐producing CD4+ T cells were quantified (n = 11 mice (control), n = 11 mice). (e) Mean fluorescent intensity (MFI) of Foxp3 protein in Treg cells in the brain at 3 weeks after EAE induction, or percentages of Treg cells among CD4+ T cells, or their absolute numbers. (f) Lymph node or splenic IL‐17A production by T cells in IL‐22 plasmid‐ or empty plasmid‐injected mice on day 7 of EAE induction (priming phase of the disease). (g) Treg cell numbers in LN and spleen on day 7 of EAE induction. The experiments were repeated at least three times. (*) indicates p‐value <0·05, (**), p < 0·01