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Thrombospondin mediates adherence of CD36+ sickle reticulocytes to
endothelial cells
K Sugihara, T Sugihara, N Mohandas and RP Hebbel
Department of Medicine, University of Minnesota Medical School,
Minneapolis.
Initiation of vasocclusion in sickle disease pathophysiology may involve
abnormal red blood cell (RBC) adhesivity to endothelium, a phenomenon
influenced by both RBC and plasma factors. Using human umbilical vein
endothelial cells and a gravity sedimentation adherence assay, we have
examined thrombospondin (TSP) as a plasma factor in this adhesive event.
The already-abnormal adherence of sickle RBCs in buffer/albumin is
significantly augmented (P < .001) by the addition of TSP, with
half-maximal effect at about 0.3 microgram/mL. This effect is abolished by
antibodies to either TSP or glycoprotein (GP) IV (CD36), as well as
peptides RGDS and CSVTCG. The even greater adherence (P < .005) of
sickle RBCs in autologous platelet-rich plasma (without added TSP) is
dramatically inhibited by alpha CD36 antibodies (OKM5 and alpha GPIV) and
significantly diminished by alpha TSP, by peptides RGDS and CSVTCG, and by
two antibodies to the vitronectin receptor (7E3 and LM609). Studies of
density-separated subpopulations and of RBC adhesion to immobilized
proteins, as well as analysis of sickle RBCs using fluorescence-activated
cell sorting and single cell microfluorometry, show that TSP responsiveness
is a feature of the immature sickle "stress" reticulocytes, which carry
CD36 (and not GPIIbIIIa-like receptors) as the TSP-receptive moiety. The
endothelial cell's participation in this phenomenon appears to be more
complex, and the data are consistent with the notion that it involves TSP
interaction with other plasma proteins and/or multiple receptor structures.
Other potential adhesogenic proteins (plasma von Willebrand factor,
vitronectin, fibrinogen, and fibronectin) neither exhibited an affinity for
reticulocytes nor supported increased sickle RBC adherence when added to
buffer/albumin in these assay systems. In aggregate, our results indicate
that TSP may be the major promoter of RBC adhesivity in plasma, and they
suggest that therapeutic benefit might derive from interference with sickle
reticulocyte CD36, as achieved by antibodies and CSVTCG in these studies.
Volume 80,
Issue 10,
pp. 2634-2642,
11/15/1992
Copyright © 1992 by The American Society of Hematology
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