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Thrombopoietin enhances the alpha IIb beta 3-dependent adhesion of
megakaryocytic cells to fibrinogen or fibronectin through PI 3 kinase
G Zauli, A Bassini, M Vitale, D Gibellini, C Celeghini, E Caramelli, S Pierpaoli, L Guidotti and S Capitani
Institute of Human Anatomy, University of Ferrara, Italy.
The effect of thrombopoietin (TPO) on the functional activity of surface
alpha IIb beta 3 (GPIIbIIIa) was investigated in both primary human
megakaryocytic cells, derived from peripheral blood CD34+ cells, and HEL
hematopoietic cell line. TPO (100 ng/mL) induced a sixfold to ninefold
enhancement of adhesion of both primary megakaryocytic and HEL cells to
plates coated with either fibrinogen or fibronectin and a parallel increase
of immunoreactivity to the PAC1 monoclonal antibody (MoAb) and fluorescein
isothiocyanate-fibrinogen, both of which recognize an activated state of
alpha IIb beta 3. The enhanced adhesion to fibrinogen or fibronectin was
mediated by the Arg-Gly-Asp (RGD) recognition sequence of alpha IIb beta 3,
as it was abolished by pretreatment of cells with saturating concentrations
of RGDS peptide. A MoAb specific for the alpha IIb beta subunit of alpha
IIb beta 3 also inhibited cell attachment to fibrinogen or fibronectin,
while MoAb to anti-alpha v beta 3 or anti-alpha 5 integrins were completely
ineffective, clearly indicating that alpha IIb beta 3 participates in this
association. A role for PI 3 kinase (PI 3-K) in the TPO-mediated increase
in alpha IIb beta 3 function in megakaryocytic cells was suggested by the
ability of the PI 3-K inhibitor wortmannin (100 nmol/L) and antisense
oligonucleotides directed against the p85 regulatory subunit of PI 3-K to
completely block the TPO-induced increase in alpha IIb beta 3 integrin
activity upon TPO stimulation. The modulation of adhesiveness to
extracellular matrix proteins containing the RGD motif mediated by TPO
likely plays a physiologic role in megakaryocytopoiesis, as pretreatment of
CD34+ cells with RGDS or anti-alpha IIb MoAb significantly reduced the
number of megakaryocytic colonies obtained in a fibrinclot semisolid assay.
Volume 89,
Issue 3,
pp. 883-895,
02/01/1997
Copyright © 1997 by The American Society of Hematology
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