Figure 8. MIF deficiency correlates with enhanced terminal RBC differentiation and reduced RBC clearance during T. brucei infection.

(A) Gating strategy used to discriminate different stages of erythroid development, starting from nucleated erythroblasts (P1 (pro), P2 (Basophilic + polychromatic) and P3 (orthochromatic)) till enucleated erythrocytes (P4 (reticulocyte) and P5 (erythrocyte)). 7AAD⁻CD45⁻Ter-119+ cells were selected and plotted in a CD44 versus FSC plot (upper panel). The percentage of the different erythroid populations at day 18 p.i. in WT (black box) and Mif ^−/− (white box) mice is shown for the spleen (B) and bone marrow (C). Results are representative of 2 independent experiments and presented as mean of 3 individual mice ± SEM. (D) RBC clearance in non-infected (WT, black bars; Mif ^−/−, white bars) and day 12 p.i. T. brucei infected (WT, dark grey bars; Mif ^−/−, light grey bars) C57Bl/6 mice following i.v injection with 200 µl GFP⁺ RBCs. At different time points after injection the presence of GFP⁺ RBCs following gating on Ter-119⁺ cells in the blood was evaluated. GFP⁺ RBC numbers were normalized, whereby GFP⁺ RBCs present after 1 day post infection are being referred as 100%. Results are representative of 2 independent experiments and presented as mean of 5 individual mice ± SEM (*: p-values ≤0.05, **: p-values ≤0.01).