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Primitive human hematopoietic progenitors adhere to P-selectin (CD62P)
AC Zannettino, MC Berndt, C Butcher, EC Butcher, MA Vadas and PJ Simmons
Leukaemia Research Unit, Hanson Centre for Cancer Research, Adelaide, South
Australia.
P-selectin was shown to bind committed human hematopoietic progenitors
(colony-forming unit-granulocyte-macrophage [CFU-GM] and burst-forming
unit-erythroid [BFU-E]) as identified by their expression of the CD34
antigen and by in vitro clonogenic assays. In addition, P-selectin bound
all precursors (pre-CFU) of committed myeloid progenitors assayed by their
ability to sustain hematopoiesis in both conventional stroma- containing
and stroma-free, cytokine-dependent systems. Binding of CD34+ cells to
P-selectin was temperature-independent and shear- resistant, occurred only
in the presence of divalent cations, was protease sensitive, and was
completely blocked by anti-P-selectin antibody. Neuraminidase treatment of
CD34+ cells completely abrogated their binding to P-selectin, implying a
prominent role for sialic acid in the structure and function of the
P-selectin ligand on hematopoietic progenitors. Monoclonal antibodies
(MoAbs) CSLEX-1 and HECA-452, which identify carbohydrate epitopes
involving sialic acid, bound to 33% and 35% of CD34+ cells, respectively,
and included the majority of CFU-GM and pre-CFU. Three-color flow
cytometric analysis showed a precise codistribution of CSLEX-1 and HECA-452
antigens on CD34+ cells, implying recognition of the same glycoprotein
antigen by the two MoAbs. Treatment of CD34+ cells with neuraminidase
completely abolished binding of both MoAbs. In addition, HECA-452 partially
blocked the adhesion of CD34+ cells to P-selectin. P-selectin glycoprotein
ligand (PSGL-1), recently molecularly cloned from the promyelocytic
leukemia cell line HL60, was expressed by CD34+ cells as determined by
reverse transcription polymerase chain reaction. Combined with the
functional and biochemical characteristics, these data suggest that PSGL-1
may comprise an important P-selectin ligand expressed by primitive
hematopoietic cells, but do not preclude the existence of additional P-
selectin ligands on these cells.
Volume 85,
Issue 12,
pp. 3466-3477,
06/15/1995
Copyright © 1995 by The American Society of Hematology
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