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The value of flow cytometric analysis of platelet glycoprotein expression
of CD34+ cells measured under conditions that prevent P- selectin-mediated
binding of platelets
MW Dercksen, IS Weimar, DJ Richel, J Breton-Gorius, W Vainchenker, CM Slaper- Cortenbach, HM Pinedo, AE von dem Borne, WR Gerritsen and CE van der Schoot
European Cancer Centre, Amsterdam, The Netherlands.
In the present study, we show by adhesion assays and ultrastructural
studies that platelets can bind to CD34+ cells from human blood and bone
marrow and that this interaction interferes with the accurate detection of
endogenously expressed platelet glycoproteins (GPs). The interaction
between these cells was found to be reversible, dependent on divalent
cations, and mediated by P-selectin. Enzymatic characterization showed the
involvement of sialic acid residues, protein(s). The demonstration of mRNA
for the P-selectin glycoprotein ligand 1 (PSGL-1) in the CD34+ cells by
polymerase chain reaction (PCR) analysis suggests that this molecule is
present in these cells. Under conditions that prevent platelet adhesion, a
small but distinct subpopulation of CD34+ cells diffusely expressed the
platelet GPIIb/IIIa complex. These cells were visualized by immunochemical
studies. Furthermore, synthesis of mRNA for GPIIb and GPIIIa by CD34+ cells
was shown using PCR analysis. The semiquantitative PCR results show
relatively higher amounts of GPIIb mRNA than of PF4 mRNA in CD34+CD41+
cells in comparison with this ratio in platelets. This finding is a strong
indication that the PCR results are not caused by contaminating adhering
platelets. MoAbs against GPIa GPIb alpha, GPV, P- selectin, and the
alpha-chain of the vitronectin receptor did not react with CD34+ cells. The
number of CD34+ cells expressing GPIIb/IIIa present in peripheral blood
stem cell (PBSC) transplants was determined and was correlated with
platelet recovery after intensive chemotherapy in 27 patients. The number
of CD34+CD41+ cells correlated significantly better with the time of
platelet recovery after PBSC transplantation (r = .83, P = .04) than did
the total number of CD34+ cells (r = .55). Statistical analysis produced a
threshold value for rapid platelet recovery of 0.34 x 10(6) CD34+CD41+
cells/kg. This study suggests that if performed in the presence of EDTA the
flow cytometric measurement of GPIIb/IIIa on CD34+ cells provides the most
accurate indication of the platelet reconstitutive capacity of the PBSC
transplant.
Volume 86,
Issue 10,
pp. 3771-3782,
11/15/1995
Copyright © 1995 by The American Society of Hematology
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