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Blood, Vol. 93 No. 5 (March 1), 1999:
pp. 1511-1523
Phenotypic and Functional Evidence for the Expression of CXCR4
Receptor During Megakaryocytopoiesis
Christel Rivière,
Frédéric Subra,
Karine Cohen-Solal,
Véronique Cordette-Lagarde,
Remi Letestu,
Christian Auclair,
William Vainchenker, and
Fawzia Louache
From the INSERM U 362, Institut Gustave Roussy; and CNRS URA 147, Institut Gustave Roussy, Villejuif, France.
The identification of stromal cell-derived factor (SDF)-1 as a
chemoattractant for human progenitor cells suggests that this chemokine
and its receptor might represent critical determinants for the homing,
retention, and exit of precursor cells from hematopoietic organs. In
this study, we investigated the expression profile of CXCR4 receptor
and the biological activity of SDF-1 during megakaryocytopoiesis. CD34+ cells from bone marrow and
cord blood were purified and induced to differentiate toward the
megakaryocyte lineage by a combination of stem-cell factor (SCF) and
recombinant human pegylated megakaryocyte growth and development factor
(PEG-rhuMGDF). After 6 days of culture, a time where mature and
immature megakaryocytes were present, CD41+ cells were
immunopurified and CXCR4mRNA expression was studied. High transcript
levels were detected by a RNase protection assay in cultured
megakaryocytes derived from cord blood CD34+ cells as
well as in peripheral blood platelets. The transcript levels were about
equivalent to that found in activated T cells. By flow cytometry, a
large fraction (ranging from 30% to 100%) of CD41+
cells showed high levels of CXCR4 antigen on their surface, its expression increasing in parallel with the CD41 antigen during megakaryocytic differentiation. CXCR4 protein was also detected on
peripheral blood platelets. SDF-1 acts on megakaryocytes by inducing
intracellular calcium mobilization and actin polymerization. In
addition, in in vitro transmigration experiments, a significant proportion of megakaryocytes was observed to respond to this chemokine. This cell migration was inhibited by pertussis toxin, indicating coupling of this signal to heterotrimeric guanine nucleotide binding proteins. Although a close correlation between CD41a and CXCR4 expession was observed, cell surface markers as well as morphological criteria indicate a preferential attraction of immature megakaryocytes (low level of CD41a and CD42a), suggesting that SDF-1 is a potent attractant for immature megakaryocytic cells but is less active on
fully mature megakaryocytes. This hypothesis was further supported by
the observation that SDF-1 induced the migration of colony forming
unit-megakaryocyte progenitors (CFU-MK) and the expression of
activation-dependent P-selectin (CD62P) surface antigen on early
megakaryocytes, although no effect was observed on mature megakaryocytes and platelets. These results indicate that CXCR4 is
expressed by human megakaryocytes and platelets. Furthermore, based on
the lower responses of mature megakaryocytes and platelets to SDF-1
as compared with early precursors, these data suggest a role for this
chemokine in the maintenance and homing during early stages of
megakaryocyte development. Moreover, because megakaryocytes are also
reported to express CD4, it becomes important to reevaluate the role of
direct infection of these cells by the human immunodeficiency virus
(HIV)-1 in HIV-1-related thrombocytopenia.
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