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Fig. 6 | Journal of Neuroinflammation

Fig. 6

From: CSF1R antagonism limits local restimulation of antiviral CD8+ T cells during viral encephalitis

Fig. 6

CSF1R antagonism does not hinder antiviral T cell recruitment to the CNS, but recruited T cells lack full activation during i.c. infection with WNV-NS5-E218A. Mice were fed PLX5622 chow or control chow for 2 weeks, then infected i.c. with WNV-NS5-E218A (104 PFU). At 8 dpi, leukocytes were isolated from the cortex and analyzed by flow cytometry. a Representative flow cytometry plots of CD45+-gated cells analyzed by CD11b vs CD45 expression, identifying three populations of cells: CD11b+CD45lo (P1), CD11b+CD45hi (P2), and CD11bnegCD45hi (P3). b Quantification of percentages and c number of cells in each gate. d Representative flow cytometry plots of CD4 vs CD8 expression and e quantification of percentages and f total numbers of CD4+ vs CD8+ cells within P3 gate. g Representative flow cytometry plots of NS4B+WNV-specific tetramer staining and h quantification of percentages and i total number of NS4B+CD8+CD45+ cells. j Representative flow cytometry plots of CD69 expression on NS4B+CD8+CD45+ cells and k quantification of percentages, l total numbers, and m MFI of CD69+NS4B+CD8+CD45+ cells. n Representative flow cytometry plots of CD160 expression on NS4B+CD8+CD45+ cells and o quantification of percentages, p total numbers, and q MFI of CD160+NS4B+CD8+CD45+ cells. MFI quantifications are shown as arbitrary units (AU) by normalizing each sample to the MFI of FMO negative control value [77]. For quantification panels, each symbol represents an individual control- (black) or PLX5622-treated (red) mouse, and bars indicate mean ± SEM. Data presented are the compilation of two independent experiments with seven to nine mice per group. Statistical significance was calculated using unpaired t tests with Welch’s correction. For all data: ns, not significant at P < 0.05; *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001

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