Erythrocyte deformability has no influence on the rate of erythrophagocytosis in vitro by autologous human monocytes/macrophages

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Abstract
Erythrocytes with decreased deformability are known to be rapidly removed from the circulation by splenic macrophages. The exact mechanism is, however, not well understood. We have analysed the phagocytosis of less-deformable erythrocytes by macrophages in vitro. Human monocytes/macrophages were isolated from peripheral blood and cultured for a total time of 6 h at 37 degrees C with 5% CO2. Autologous erythrocytes of the rhesus positive donor were rigidified by heat treatment (47 degrees C for 1 h). The change in erythrocyte deformability was assessed with a filter aspiration technique; the membrane elastic modulus was found to be increased about 2.5-fold. For controls, untreated erythrocytes and erythrocytes incubated with anti-RhD-antibodies were prepared. The rate of phagocytosis during 2 h at 37 degrees C and 5% CO2 was 0.74 +/- 0.59 (erythrocytes per monocyte/macrophage) for controls, 3.58 +/- 2.72 for anti-RhD-loaded erythrocytes and 0.82 +/- 0.74 for heat-treated erythrocytes, respectively. We conclude that decreased erythrocytes deformability does not cause an increased rate of phagocytosis by monocytes/macrophages compared to normally deformable erythrocytes in our in vitro model. This suggests that the preferential removal of rigid cells in vivo is probably not a specific process, but is due to the increased splenic transit time of rigid erythrocytes and hence longer interaction time between erythrocytes and phagocytes.