Fig. 3
From: Inflammatory monocytes and microglia play independent roles in inflammatory ictogenesis
Viral load in the brain is not reduced in the low inoculum group, but inflammatory chemokine and cytokine production are reduced by more than 90% at 1 dpi. A Viral load was assessed at 1, 3, and 7 dpi in mice inoculated with 200,000 PFU or 12,500 PFU of TMEV. Despite a 16-fold reduction in the amount of virus delivered to the brain, the groups exhibit similar viral loads at each timepoint. F(5,70) = 0.1534, P = 0.6964 between 200,000 PFU and 12,500 PFU by two-way ANOVA on ranks (inoculum x day); Shapiro–Wilk P = 0.0033; Tukey HSD pairwise analysis on ranks: 200,000 PFU vs 12,500 PFU @ 1 dpi: P = 0.6938; @ 3 dpi: P = 0.9900; @ 7 dpi: P = 0.9989; results from 5 separate experiments. B, C Representative images of TMEV immunostaining (green) in CA1 pyramidal neurons at 3 dpi in a mouse inoculated with 200,000 PFU (B) or 12,500 PFU (C) of virus; DAPI is shown in blue. Boxes indicate higher magnification insets “i” and “ii”. The amount of CCL2 (D), TNFα (E), and IL6 (F) in hippocampal homogenates was measured at 24 and 72 hpi. All of the factors were profoundly reduced at 24 hpi in the mice inoculated with 12,500 PFU of TMEV relative to mice inoculated with 200,000 PFU, with TNFα and IL6 indistinguishable from vehicle control mice. D CCL2, F(2,12) = 318.3, P < 0.0001 by two-way ANOVA; Shapiro–Wilk P = 0.8826 (24 hpi); Tukey’s pairwise analysis: 24 hpi: 200,000 PFU vs 12,500 PFU: P < 0.0001; 200,000 PFU vs vehicle: P < 0.0001; 12,500 PFU vs vehicle: P = 0.5123; 72 hpi: 200,000 PFU vs 12,500 PFU: P = 0.4512; 200,000 PFU vs vehicle: P = 0.9999; 12,500 PFU vs vehicle: P = 0.5252; ds @ 24 hpi = 11.8; results from 3 separate experiments. E TNFα, F(2,28) = 15.92, P < 0.0001 by two-way ANOVA; Shapiro–Wilk P = 0.7674; Tukey’s pairwise analysis: 24 hpi: 200,000 PFU vs 12,500 PFU: P < 0.0001; 200,000 PFU vs vehicle: P < 0.0001; 12,500 PFU vs vehicle: P > 0.9999; 72 hpi: 200,000 PFU vs 12,500 PFU: P = 0.1562; 200,000 PFU vs vehicle: P = 0.3342; 12,500 PFU vs vehicle: P > 0.9999; ds @ 24 hpi = 4.6; results from 3 separate experiments. F IL6, F(2,28) = 28.975, P < 0.0001 by two-way ANOVA; Shapiro–Wilk P = 0.5052; Tukey’s pairwise analysis: 24 hpi: 200,000 PFU vs 12,500 PFU: P < 0.0001; 200,000 PFU vs vehicle: P < 0.0001; 12,500 PFU vs vehicle: P = 0.9993; 72 hpi: 200,000 PFU vs 12,500 PFU: P = 0.9975; 200,000 PFU vs vehicle: P = 0.9914; 12,500 PFU vs vehicle: P > 0.9999; ds @ 24 hpi = 4.15; results from 3 separate experiments. Scale bar in C is 200 μm and refers to B; scale bar in i is 100 μm and refers to i