Figure 5
In vivo IL-30 administration develops significantly exacerbated EAE. C57BL/6 mice were induced for EAE. Osmotic pumps containing IL-27 (400 ng, n = 5), IL-30 (400 ng, n = 5) were subcutaneously implanted or sham surgery (n = 4) was performed at 12 days post induction. (a) EAE score. (b) Total CD4+ T cell numbers in the CNS at 22 days post induction. (c) The numbers of GM-CSF, IFNγ, IL-17, and TNFα+ CD4 T cells were determined by intracellular cytokine staining at day 22 post immunization. (d) qPCR analysis of the indicated chemokine expression in the brain from the sham, IL-27-pump and IL-30-pump group. Gene expression was normalized by Gapdh and compared to that of sham surgery group. (e) Lag3 expression of CNS infiltrating Treg cells was determined by flow cytometry. (f) Cx3cr1WT and Cx3cr1ΔIl27p28 mice induced for EAE as described in Fig. 4 were used to measure CNS infiltrating Treg cell expression of Lag3. (g,h) IL-30 and IL-27 levels in the CNS homogenates were measured by ELISA and were normalized to those of naïve wild type mice. n = 7 per group. *p < 0.05; **p < 0.01; ***p < 0.001; as determined by Mann–Whitney nonparametric test.
