Fig. 5
IL-10R antagonist reverses the effect of ICOSaa treatment in mice treated with paclitaxel. A Schematic representation of experimental design. Arrow represents days with von Frey testing. Groups are shown as: Vehicle + IL-10Ra (grey), Ptx + IgG (yellow), Ptx + ICOSaa + IgG (blue), Ptx + ICOSaa + IL-10Ra (pink), Ptx + IL-10Ra [32]. B Mechanical nociceptive thresholds are shown for each group with statistical differences represented in the graph (two-way ANOVA, F = 1.921, p-value = 0.0048, post hoc Bonferroni, Vehicle + IL-10Ra vs. Ptx + IgG, p-value = 0.0362, Vehicle + IL-10Ra vs. Ptx + ICOSaa + IgG, p-value = ns, Vehicle + IL-10Ra vs. Ptx + IL-10Ra, p-value = 0.0485, Vehicle + IL-10Ra vs. Ptx + ICOSaa + IL-10Ra, p-value = 0.0452, $Ptx + IgG vs Ptx + ICOSaa + IgG, p-value = 0.0574, Ptx + IgG vs Ptx + IL-10Ra, p-value = ns, Ptx + IgG vs. Ptx + ICOSaa + IL-10Ra, p-value = ns, #Ptx + ICOSaa + IgG vs. Ptx + IL-10Ra, p-value = 0.0697, *Ptx + ICOS + IgG vs. Ptx + ICOS + IL-10Ra, p-value = 0.0697, Ptx + ICOSaa + IL-10Ra vs. Ptx + IL-10Ra, p-value = ns). N = 4, 5, 5, 5, 5 per group, respectively. C The effect size was significant between Ptx + ICOSaa + IL-10Ra and Ptx + ICOSaa + IgG1 (one-way ANOVA, F = 7.211, p-value = 0.0010, post hoc Tukey, Vehicle + IL-10Ra vs. Ptx + IgG, p-value = 0.0108, Vehicle + IL-10Ra vs. Ptx + ICOSaa + IgG, p-value = ns, Vehicle + IL-10Ra vs. Ptx + IL-10Ra p-value = 0.0173, Vehicle + IL-10Ra vs. Ptx + ICOSaa + IL-10Ra, p-value = 0.0079, Ptx + IgG vs. Ptx + ICOSaa + IgG, p-value = 0.0301, Ptx + IgG vs. Ptx + IL-10Ra, p-value = ns, Ptx + ICOSaa + IgG vs.Ptx + IL-10Ra, p-value = 0.0485, Ptx + IL-10Ra + IgG vs.Ptx + ICOSaa + IL-10Ra, p-value = ns). D A significant increase in IL-10 expression was observed in mice subjected to intrathecal injection of ICOSaa using ELISA (one-way ANOVA, F = 8.845, p-value = 0.0075, post hoc Tukey, Vehicle vs. Ptx, p-value = ns, Vehicle vs. Ptx + ICOSaa, p-value = 0.0079, Ptx vs. Ptx + ICOSaa, p-value = 0.0293) N = 4/group. E Representative flow cytometry plots of subset of T cells CD4-positive and CD8-positive (previously gated on CD45posCD3pos) in each group after treatment, isolated on day 13 from L3–L5 DRG. F Percentage of CD4-positive T cells was increased in Ptx and Ptx + ICOSaa groups compared to vehicle treated cohort and no significant changes were observed between Ptx and Ptx + ICOSaa groups (one-way ANOVA, F = 10.86, p-value = 0.004, post hoc Tukey, Vehicle vs. Ptx,p-value = 0.0073, Vehicle vs. Ptx + ICOSaa, p-value = 0.076, Ptx vs. Ptx + ICOSaa, p-value = ns) N = 4/group. G Percentage of CD8-positive T cells were increased only in Ptx + ICOSaa group compared to vehicle control (one-way ANOVA, F = 4.881, p-value = 0.0367, post hoc Tukey, Vehicle vs. Ptx, p-value = ns, Vehicle vs. Ptx + ICOSaa, p-value = 0.0301, Ptx vs. Ptx + ICOSaa p-value = ns), N = 4/group, *#$p < 0.05