Membrane damages under high pressure of human erythrocytes agglutinated by concanavalin A

Human erythrocytes are agglutinated by lectins such as concanavalin A (Con A). The behaviors of agglutinated erythrocytes under pressure are less well understood. Here, we report the effects of erythrocyte agglutination on pressure-induced membrane damages. Small clumps of intact erythrocytes by Con A were dissociated by a pressure of 200 MPa. Further, the observation by scanning electron microscopy demonstrated the generation of vesicles, fragmented particles, and membrane hole. On the other hand, large clumps of trypsin-digested erythrocytes by Con A seemed to be stable against 200 MPa. However, the erythrocytes dissociated from such pressure-treated clumps by methyl α-mannopyranoside also showed the existence of vesicles and fragmented particles except for the membrane hole. Pressure-induced hemolysis was greatly suppressed in such large clumps. Similar suppressive effects were observed in erythrocytes packed by centrifugation. However, the hemolysis occurred when the erythrocytes dissociated from 200 MPa-treated large clumps by methyl α-mannopyranoside were incubated at 0°C and atmospheric pressure. Pyrene excimer fluorescence due to spectrin denaturation was observed in Con A-agglutinated ghosts that were exposed to a pressure of 200 MPa. These results suggest that upon pressure treatment of tightly agglutinated erythrocytes, the hemolysis is greatly suppressed, but membrane damages occur such as spectrin denaturation and vesiculation.