Figure 4.

Asymptomatic HSV-1 infection increases spinal cord demyelination after EAE induction. Mice were treated with mock, infected with WT HSV-1 (17syn+ strain) or inoculated with HSV-1 Δ34.5 (F strain) (n = 12/group). EAE was induced 30–35 days post-treatment and 4 mice from each group were euthanized at day 14, 21 or 25 post-EAE induction. (A) Representative images of lumbar sections of spinal cords stained with Luxol Fast Blue showing tissue demyelination. (B) Representative images of Luxol Fast Blue staining contrasted with Cresyl violet showing cellular infiltration. Myelin staining is observed in blue in the white matter and cell nuclei are colored purple. (C) Representative images of immunohistochemistry performed against the MBP protein. Image magnifications are 10× (left) and 20× (right) and correspond to day 21 post-EAE induction. (D) Quantitative histopathological analyses of spinal cord lumbar sections. Values represent the mean ± SEM of three independent experiments (4 mice/group per day evaluated). Data were analyzed using one-way ANOVA with Dunnett's multiple comparison post-test; **p < 0.01; n.s. non-significant. (E) Representative western blot images for MBP (upper panel) and β-actin (lower panel) in the spinal cord at day 14 post-EAE induction. The graph shows densitometric analyses for MBP bands that were normalized to β-actin. Data represent the mean ± SEM of two independent experiments (n = 6). Comparisons between ratios were performed using one-way ANOVA with Dunnett's multiple comparison post-test; *p < 0.05.