Figure 3
PbNK65F- and PbNK65S-infected mice have similar quantities of CD8+ T cells and iRBCs in their brains. (A, B) Mice were infected as indicated in Fig. 1. At days 10 or 21 post infection mice were injected intravenously with AF-488 conjugated CD45 specific Abs (in vivo) to label intravascular leukocytes. Brains were harvested shortly after and processed as detailed in methods section. Lymphocytes were then incubated with a second Ab including BV421 labelled CD45 specific Abs (in vitro) which stains both vascular and extravascular leukocytes. CD11b was used to gate out microglia and CD8 was used to gate CD8+ T cells. Within the CD8+ T cell gate cells that are double positive for both CD45 stains are gated as intraluminal cells (purple gate) and cells that are single positive for only in vitro CD45 stain were gated as adluminal cells (green gate) (A). Absolute counts of parenchymal CD8+ T cells are graphed (B). Each circle represents an individual mouse n.s. = P > 0.05 (One way ANOVA with Dunnet’s multiple comparisons test). (C) qPCR comparisons of parasite levels in brain specimens taken at day 21 post infection from mice infected as indicated in Fig. 1. Each circle represents an individual mouse. n.s. = P > 0.05 (Welch’s T-test).
