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. 2021 Sep 7;12:700501. doi: 10.3389/fimmu.2021.700501

Figure 5.

Figure 5

The presence of L. donovani parasites in the BM is necessary to maintain monocyte activation. (A–E) B6.Il10-/- mice were re-infected (RI) at d56 with 3x107 L. donovani amastigotes and killed 1 day post re-infection. (A) Experimental strategy. (B) Parasite load per one million cells determined by limiting dilution assay in the BM at day 56 or 1 day post RI. Mann-Whitney test applied: P=0.006 (**); 95% CI: 87.73 – 200; statistical power: 100%; n=7. (C) Percentage of MHCII on Ly6C+CCR2+ monocytes in the BM. Brown-Forsythe ANOVA applied only to day 56 data: P=0.0019; statistical power: 99.5%; n≥3; post hoc test Dunnett’s T3: statistical difference reached in B6.wt vs. B6.il10-/- (P=0.0073; **); 95% CI: 5.06 – 44.58) and B6.il10-/- vs. B6.il10-/- RI (P=0.0024; **); 95% CI: -51.32 – -12.13). (D) Level of MHCII expressed by the BM Ly6C+CCR2+ monocytes represented on histogram. (E) MFI of MHCII on Ly6C+CCR2+ monocytes in the BM of the indicated mice. Brown-Forsythe ANOVA applied only to day 56 data: P=0.0019; statistical power: 26.7%; n≥3; post hoc test Dunnett’s T3: statistical difference reached in B6.wt vs. B6.il10-/- RI (P=0.0006; ***); 95% CI: -4297 – -1310) and B6.il10-/- vs. B6.il10-/- RI (P=0.0002: ***); 95% CI: -5099 – -2266). (F–H) B6 mice infected for 28 days were treated with 10mg/kg of Ambisome®. Data were collected 3 days post-treatment. (F) Parasite load measured by limiting dilution assay in the BM. Welch’s t test applied in (F) P<0.0001; ****; 95% CI: -3.31 – -1.67; statistical power: 100%; n≥6. Percentage (G) and MFI (H) of MHCII expressed on Ly6C+CCR2+ monocytes. Brown-Forsythe ANOVA applied in (G): P<0.0001; statistical power: 100%; n≥3; post hoc test Dunnett’s T3: statistical difference reached in all pairs (P<0.0001; ****); and Kruskal-Wallis test applied in (H): P=0.0002; statistical power: 100%; n≥3; post hoc test Dunn’s MCT in Na+Amb vs. Inf (P=0.0071; **). In (A–E), data are from 3 to 5 mice per group/experiment. Experiments were performed twice and pooled data are shown. In (F–H), data were derived from 3 to 4 mice/group. Mean ± SD is shown except in A Median with 95% CI is shown.