Figure 5

Increased production of INF-γ and TNF-α by CD8⁺ T cells of N. caninum infected mice.

(a) Percentage of splenic CD8-gated T cells expressing IFN-γ or TNF-α of infected mice (NcT) and controls (PBS), detected by intracellular staining after stimulation with PMA/ionomycin. Mean fluorescence intensities due to respective cytokine staining are also presented. Bars represent means plus one SD of pooled data from two independent experiments (n = 6 for controls and 4-day infected mice and n = 9 for 7-day infected mice). Unpaired two-tailed t-test was used to compare parasite-inoculated vs respective control mouse groups. Statistical significance between infected mice and controls is indicated above bars. (b) Percentage of IFN-γ⁺ cells on total CD8⁺ T cells of infected mice (NcT) and controls (PBS) detected in in vitro splenocytes cultures non-stimulated (−) or stimulated for 16 h with N. caninum sonicates (+); n = 5 and n = 7 for non-stimulated and stimulated groups, respectively. Contour plots correspond to a representative example of CD8-gated T cells of the analysed samples. Analysis regions were set according to isotype control-stained samples. Numbers within contour plots correspond to the percentage of cells in the region shown. (c) IFN-γ concentration in the supernatants of splenocyte cultures non-stimulated (−) or stimulated for 16 h with N. caninum sonicates (+); n = 5 and n = 7 for non-stimulated and stimulated groups, respectively; BDL-below detection limit (15 pg/ml). Statistical significances between indicated pair groups on panels (b) and (c) were determined by one-way ANOVA and Tukey’s post-hoc test and are shown above bars.