Figure 6.
Clearance kinetics of SMEs in clodronate-treated mice. (A) Representative 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 of a clodronate-treated 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 clodronate-treated mice (n = 10 per group). (C) Posttransfusion recovery of long-stored RBCs is increased at 2 and 24 hours after transfusion to clodronate-treated 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 clodronate-treated mice (n = 10 per group). (E) Decreased proportion of long-stored RBCs that were cleared 60 to 240 minutes posttransfusion in clodronate-treated 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 spleen (red line), liver (orange line), and bone marrow (blue line) after transfusion in clodronate-treated mice (n = 3 mice per time point). (G) Posttransfusion erythrophagocytosis of RBCs in spleen (i) and liver (ii), estimated by the increase in MFI of CFSE in monocytes (blue lines), inflammatory monocytes (purple lines), and granulocytes (orange lines) in clodronate-treated recipients compared with control nontransfused mice (n = 3 mice per time point). Data are presented as means ± standard errors of the mean. **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 clodronate-treated 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 clodronate-treated mice. (A) Representative 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 of a clodronate-treated 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 clodronate-treated mice (n = 10 per group). (C) Posttransfusion recovery of long-stored RBCs is increased at 2 and 24 hours after transfusion to clodronate-treated 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 clodronate-treated mice (n = 10 per group). (E) Decreased proportion of long-stored RBCs that were cleared 60 to 240 minutes posttransfusion in clodronate-treated 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 spleen (red line), liver (orange line), and bone marrow (blue line) after transfusion in clodronate-treated mice (n = 3 mice per time point). (G) Posttransfusion erythrophagocytosis of RBCs in spleen (i) and liver (ii), estimated by the increase in MFI of CFSE in monocytes (blue lines), inflammatory monocytes (purple lines), and granulocytes (orange lines) in clodronate-treated recipients compared with control nontransfused mice (n = 3 mice per time point). Data are presented as means ± standard errors of the mean. **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 clodronate-treated 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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