Fig. 5.

Leukocyte accumulation in the CNS of viperin^−/− mice after WNV infection. Wild-type and viperin^−/− mice were inoculated with 10² PFU of WNV by subcutaneous injection into the footpad. Brains (A to D) and spinal cords (E to H) were harvested on days 9 and 10, respectively, and leukocytes were isolated by Percoll gradient centrifugation. (A and E) The total number of CD3⁺ CD4⁺ or CD3⁺ CD8⁺ T cells was determined by multiplying the percentage of CD3⁺ CD8⁺ cells by the total cell count. (B and C) Brain leukocytes were stimulated ex vivo with a D^b-restricted NS4b WNV peptide; stained for CD3 and CD8 and intracellular granzyme B, IFN-γ, or TNF-α; and analyzed by flow cytometry. (F and G) Spinal cord leukocytes were stained directly with D^b-NS4b tetramers and antibodies to CD8 and granzyme B. (D and H) The numbers of macrophages (CD11b^high/CD45^high) and microglia (CD11b^high/CD45^low) were also measured. Differences were not statistically significant, and data represent the averages of data from 3 to 4 independent experiments with 3 to 5 mice per experiment (n = 11 to 19).