Figure 6. Guanabenz treatment protects oligodendrocytes and alters CD4+ T cell populations in two EAE mouse models.

Flow cytometry analysis of immune cell populations in the (a) inguinal lymph nodes, (b) spleen and (c) CNS, taken from PID15 mice with actively induced chronic EAE treated daily with vehicle or guanabenz beginning PID7. Data are representative of four mice per group; experiment performed twice. (d,e) Analysis of IFN-γ and IL-17 protein expression in the lumbar spinal cord of PID9 and PID15 chronic EAE mice. PID9, n=4–5 mice per group, PID15, n=6–9 mice per group. (f–o) Flow cytometry analysis of adoptive transfer EAE mice. Recipient mice were treated daily with vehicle or guanabenz beginning PCT day 0. C57BL/6 mice that received no blast cells and only two pertussis toxin treatments (Ptx only) were included as flow analysis controls. (f) All mice were followed for EAE disease severity until euthanized, with five mice in each group remaining on day 16. The last guanabenz treatment was on day 9, and the subsequent rapid clinical decline of these animals demonstrated that they were the recipients of active T cells and the protective nature of guanabenz. On days 3, 6 and 10 the CNS was collected from four representative mice in each treatment group, and the number of (g) total live CD4+ T cells, (h) dead CD4+ T cells, (i) AnnexinV+ CD4+ T cells, (j) Ki67+ CD4+ T cells, (k) CD44hi CD4+ T cells, (l) IFN-γ+ CD4+ Th1 cells and (m) IL-17+ CD4+ Th17 cells was assessed via flow cytometry. The number of (n) live mature GALC+ MOG+ oligodendrocytes present within the CNS and the number of (o) A2B5+ PDGFRα+ early progenitor OPCs were also assessed via flow cytometry. The data are presented as the average number of cells over time. *P<0.05, **P<0.005, ***P<0.0005, as compared with vehicle-treated mice. Data represents average of four mice per group, presented as mean±s.e.m.