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Blood, Vol. 95 No. 4 (February 15), 2000:
pp. 1293-1300
Adherence of phosphatidylserine-exposing erythrocytes to
endothelial matrix thrombospondin
Annamaria B. Manodori,
Gilda A. Barabino,
Bertram
H. Lubin, and
Frans A. Kuypers
From the Children's Hospital Oakland Research Institute, Oakland,
CA; and Department of Chemical Engineering, Northeastern University,
Boston, MA.
Phospholipid asymmetry is well maintained in erythrocyte (RBC)
membranes with phosphatidylserine (PS) exclusively present in the inner
leaflet. The appearance of PS on the surface of the cell can have major
physiologic consequences, including increased cell-cell interactions.
Because increased adherence of PS-exposing RBCs to endothelial cells
(ECs) may be pathologically important in hemoglobinopathies such as
sickle cell disease and thalassemia, we studied the role of PS exposure
in calcium ionophore-treated normal RBC adherence to human umbilical
vein endothelial cell (HUVEC) monolayers. When HUVEC monolayers were
incubated with these PS-exposing RBCs, the ECs retracted and the RBCs
adhered primarily in the gaps opened between the ECs. A linear
correlation was found between the number of PS-exposing RBCs in
the population and the number of adhering RBCs to the monolayer.
Pretreatment of RBCs with annexin V significantly decreased
adherence by shielding PS on the RBCs. Similarly, PS-containing lipid
vesicles decreased RBC binding by competing for the PS binding sites in
the monolayer. PS-exposing RBCs and PS-containing lipid vesicles
adhered to immobilized thrombospondin (TSP) and matrix TSP,
respectively, and adherence of PS-exposing RBCs to EC monolayers was
reduced by antibodies to TSP and to its EC receptor,
 v 3. Together, these results indicate a
role for PS and matrix TSP in the adherence of PS-exposing RBCs to EC
monolayers, and suggest an important contribution of PS-exposing RBCs
in pathologies with reported vascular damage, such as sickle cell anemia.
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