Figure 7.
Clearance kinetics of SMEs in splenectomized mice. (A) Typical normalized frequency plots of projected surface area for long-stored mouse RBCs, as observed 5 minutes (green line), 2 hours (yellow line), and 24 hours (red line) after transfusion to a splenectomized mouse. Control fresh RBCs from a nontransfused mouse (blue) are shown as reference. Dashed black vertical line defines gating of SME. (B) Delayed clearance of SMEs in circulation after transfusion in splenectomized mice (n = 10 per group). (C) Posttransfusion recovery of long-stored RBCs is increased at 24 hours after transfusion to splenectomized recipients (dashed line; n = 10) compared with controls (solid line; n = 10). (D) Variable persistence in circulation of long-stored RBCs (lavender dashed line) that contain the 2 complementary subpopulations of SMEs (black dotted line) and morphologically normal RBCs (light blue solid line), computed by combining flow cytometric and Imagestream data, after transfusion in splenectomized mice (n = 10 per group). (E) Decreased proportion of long-stored RBCs that were cleared at 240 minutes posttransfusion in splenectomized recipients (dashed line; n = 6 mice per time point) compared with controls (solid line; n = 8 mice per time point). (F) EF (ratio of transfused CFSE+ RBCs in sliced organ/CFSE+ RBCs in venous blood) 5 to 240 minutes posttransfusion in liver (orange line) and bone marrow (blue line) after transfusion in splenectomized mice (n = 3 mice per time point). (G) Posttransfusion erythrophagocytosis of RBCs in liver (i) and bone marrow (ii), estimated by the increase in MFI of CFSE in macrophages (red lines), monocytes (blue lines), inflammatory monocytes (purple lines), and granulocytes (orange lines) in splenectomized recipients compared with control nontransfused mice (n = 3 mice per time point). Data are presented as means ± standard errors of the mean. *P < .05, **P < .01, ***P < .001, ****P < .0001 by Kruskal-Wallis test compared with fresh RBC condition (B), by Sidak multiple comparisons test comparing, at each time point, recovery (clearance) in splenectomized vs control recipients (C,E), and by Sidak multiple comparisons test comparing, at each time point, recovery of SME subpopulation vs normal subpopulation (D).

Clearance kinetics of SMEs in splenectomized mice. (A) Typical normalized frequency plots of projected surface area for long-stored mouse RBCs, as observed 5 minutes (green line), 2 hours (yellow line), and 24 hours (red line) after transfusion to a splenectomized mouse. Control fresh RBCs from a nontransfused mouse (blue) are shown as reference. Dashed black vertical line defines gating of SME. (B) Delayed clearance of SMEs in circulation after transfusion in splenectomized mice (n = 10 per group). (C) Posttransfusion recovery of long-stored RBCs is increased at 24 hours after transfusion to splenectomized recipients (dashed line; n = 10) compared with controls (solid line; n = 10). (D) Variable persistence in circulation of long-stored RBCs (lavender dashed line) that contain the 2 complementary subpopulations of SMEs (black dotted line) and morphologically normal RBCs (light blue solid line), computed by combining flow cytometric and Imagestream data, after transfusion in splenectomized mice (n = 10 per group). (E) Decreased proportion of long-stored RBCs that were cleared at 240 minutes posttransfusion in splenectomized recipients (dashed line; n = 6 mice per time point) compared with controls (solid line; n = 8 mice per time point). (F) EF (ratio of transfused CFSE+ RBCs in sliced organ/CFSE+ RBCs in venous blood) 5 to 240 minutes posttransfusion in liver (orange line) and bone marrow (blue line) after transfusion in splenectomized mice (n = 3 mice per time point). (G) Posttransfusion erythrophagocytosis of RBCs in liver (i) and bone marrow (ii), estimated by the increase in MFI of CFSE in macrophages (red lines), monocytes (blue lines), inflammatory monocytes (purple lines), and granulocytes (orange lines) in splenectomized recipients compared with control nontransfused mice (n = 3 mice per time point). Data are presented as means ± standard errors of the mean. *P < .05, **P < .01, ***P < .001, ****P < .0001 by Kruskal-Wallis test compared with fresh RBC condition (B), by Sidak multiple comparisons test comparing, at each time point, recovery (clearance) in splenectomized vs control recipients (C,E), and by Sidak multiple comparisons test comparing, at each time point, recovery of SME subpopulation vs normal subpopulation (D).

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