TY - JOUR
T1 - The value of flow cytometric analysis of platelet glycoprotein expression on CD34+ cells measured under conditions that prevent P-selectin-mediated binding of platelets
AU - Wouter Dercksen, M.
AU - Weimar, Iris S.
AU - Richel, Dick J.
AU - Breton-Gorius, Janine
AU - Vainchenker, William
AU - Slaper-Cortenbach, Ineke C.M.
AU - Pinedo, Herbert M.
AU - Von Dem Borne, Albert E.G.Kr
AU - Gerritsen, Winald R.
AU - Van Der Schoot, C. Ellen
N1 - Copyright:
Copyright 2020 Elsevier B.V., All rights reserved.
PY - 1995/11/15
Y1 - 1995/11/15
N2 - 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) containing O-linked glycans, and elastase- sensitive 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, GPIbα, GPV, P-selectin, and the α-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 to 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 106 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.
AB - 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) containing O-linked glycans, and elastase- sensitive 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, GPIbα, GPV, P-selectin, and the α-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 to 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 106 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.
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U2 - 10.1182/blood.v86.10.3771.bloodjournal86103771
DO - 10.1182/blood.v86.10.3771.bloodjournal86103771
M3 - Article
C2 - 7579344
AN - SCOPUS:0028863496
SN - 0006-4971
VL - 86
SP - 3771
EP - 3782
JO - Blood
JF - Blood
IS - 10
ER -