Effects of macrophage depletion on the expression of IL-12p35, IL-12p40, CD4, CD8, GFAP, and IFN-γ transcripts in brains and spinal cords of HSV-IL-2 infected mice on day 10 PI.

<p>Female WT mice with or without macrophage depletion were ocularly infected with HSV-IL-2 or parental virus as in <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1006401#ppat.1006401.g004" target="_blank">Fig 4</a>. The brains and spinal cords were isolated on day 10 PI and qRT-PCR was performed using total RNA extracted from the individual brains. GAPDH expression was used to normalize the relative expression of each transcript. The expression of each transcript in the brains and spinal cords of naive WT mice was determined and used to estimate the relative expression of each transcript in the brains and spinal cords of the ocularly infected mice. Each point represents the mean ± SEM from 5 brains. Asterisks (*) indicate p < 0.05 by t-test. Each point represents the mean ± SEM from 5 brains or 5 spinal cords. Asterisks (*) indicate p < 0.05 by t-test. Panels: A) IL12p35 transcript in brain and spinal cord of infected mice; B) IL12p40 transcript in brain and spinal cord of infected mice; C) CD4 transcript in brain and spinal cord of infected mice; D) CD8 transcript in brain and spinal cord of infected mice; E) IFN-γ transcript in brain and spinal cord of infected mice; and F) GFAP transcript in brain and spinal cord of infected mice.</p

Role of macrophages in HSV-induced CNS demyelination.

<p>Female FoxP3^DTR mice were depleted of macrophages or mock-depleted and infected ocularly with HSV-IL-2 or parental virus (dLAT2903) (5 mice per group). Optic nerve, brain, and spinal cord were collected from euthanized mice on day 14 PI and post-fixed tissue sections were stained with LFB. Representative photomicrographs are shown (Magnification, 20_×; Size bar, 100 μm). Arrows indicate the areas of demyelination (a plaque). Panels: A) No depletion, HSV-IL-2 infected optic nerve; B) No depletion, HSV-IL-2 infected brain; C) No depletion, HSV-IL-2 infected spinal cord; D) No depletion, parental virus infected optic nerve; E) No depletion, parental virus infected brain; F) No depletion, parental virus infected spinal cord; G) Macrophage depletion, HSV-IL-2 infected optic nerve; H) Macrophage depletion, HSV-IL-2 infected brain; I) Macrophage depletion, HSV-IL-2 infected spinal cord; J) Macrophage depletion, parental virus infected optic nerve; K) Macrophage depletion, parental virus infected brain; and L) Macrophage depletion, parental virus infected spinal cord.</p

Role of FoxP3-expressing cells in HSV-induced CNS demyelination.

<p>Female FoxP3^DTR mice were depleted of FoxP3-expressing cells or depleted of both FoxP3-expressing cells and macrophages then ocularly infected with HSV-IL-2 or parental virus as in <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1006401#ppat.1006401.g001" target="_blank">Fig 1</a>. Optic nerve, brain, and spinal cord were collected from euthanized mice on day 14 PI and post-fixed tissue sections were stained with LFB. Representative photomicrographs are shown (Magnification, 20_×; Size bar, 100 μm). Arrows indicate the areas of demyelination. Panels: A) FoxP3 depleted, HSV-IL-2 infected optic nerve; B) FoxP3 depleted, HSV-IL-2 infected brain; C) FoxP3 depleted, HSV-IL-2 infected spinal cord; D) FoxP3 depleted, parental virus infected optic nerve; E) FoxP3 depleted, parental virus infected brain; F) FoxP3 depleted, parental virus infected spinal cord; G) FoxP3 and Macrophage depleted, HSV-IL-2 infected optic nerve; H) FoxP3 and Macrophage depleted, HSV-IL-2 infected brain; I) FoxP3 and Macrophage depleted, HSV-IL-2 infected spinal cord; J) FoxP3 and Macrophage depleted, parental virus infected optic nerve; K) FoxP3 and Macrophage depleted, parental virus infected brain; and L) FoxP3 and Macrophage depleted, parental virus infected spinal cord.</p

Severity of CNS demyelination in macrophage recipient mice.

<p>The LFB-stained sections of the brains, spinal cords, and optic nerves of WT mice that received adoptively transferred macrophages from WT mice and analyzed as described in <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1006401#ppat.1006401.t002" target="_blank">Table 2</a> were further analyzed in terms of the size of the demyelination plaques in the entire sections of brain, spinal cord, and optic nerves. Data are presented as mean demyelination using a total of 150 sections for brain and spinal cord and 30 sections for optic nerve from 5 mice per group. Arrows indicate no demyelination in the brain, spinal cord and optic nerve of mice received 1 X 10⁷ macrophages infected with HSV-IL-12p70 virus.</p

Comparison of mRNA expression profiles of selected genes between macrophage-intact mice infected with HSV-IL-2 or parental virus on day 5 PI^a.

<p>Comparison of mRNA expression profiles of selected genes between macrophage-intact mice infected with HSV-IL-2 or parental virus on day 5 PI<a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1006401#t001fn001" target="_blank">^a</a>.</p

Effect of adoptive transfer of macrophages on blocking CNS demyelination in HSV-IL-2 infected mice^a.

<p>Effect of adoptive transfer of macrophages on blocking CNS demyelination in HSV-IL-2 infected mice<a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1006401#t002fn001" target="_blank">^a</a>.</p