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CD64/Fc gamma RI is a granulo-monocytic lineage marker on CD34+
hematopoietic progenitor cells
J Olweus, F Lund-Johansen and LW Terstappen
Becton Dickinson Immunocytometry Systems, San Jose, CA 95131-1807, USA.
The aim of this study was to identify markers specific for granulo-
monocytic commitment of progenitor cells. Large panels of antibodies were
screened for selective staining of subsets of CD34+ cells from fetal and
adult bone marrow. Flow cytometric analysis showed that CD64/fc gamma RI
was undetectable on noncommitted progenitor cells (CD34++, CD38-/lo,
HLA-DR+) and expressed on a subset of lineage- committed progenitors
(CD34+, CD38+) with higher mean orthogonal light scatter than the remaining
CD34+ cells. The CD34+, CD64+ cells were CD19- and the majority were
CD45RA+, CD71lo, suggesting that CD64 recognized granulomonocytic
progenitor cells. Specificity of CD64 for the granulo-monocytic lineage was
shown by demonstrating that colonies arising from CD34+, CD64+ cells
consisted of 98% +/- 2% colony-forming unit-granulocyte-macrophage (CFU-GM)
in semisolid medium containing stem cell factor (SCF), interleukin-3
(IL-3), IL-6, granulocyte- macrophage colony-stimulating factor (GM-CSF),
and erythropoietin (EPO). In contrast, 63% +/- 15% of the colonies from the
CD34+, CD64- cells were burst-forming unit-erythroid/colony-forming
unit-erythroid (BFU-E/CFU-E). Furthermore, four-color immunofluourescence
and cell sorting was used to analyze the progeny of cells cultured in
liquid medium containing identical cytokines as used in the semisolid
medium. This analysis showed that CD34+, CD64+ cells gave rise to 83% +/-
10% granulo-monocytic cells whereas progeny of the CD34+, CD64- cells
contained 81% +/- 11% erythroid cells. Neutrophils as well as basophils and
monocytes/macrophages were present in the cultures from CD34+, CD64+ cells,
showing that this population contains progenitors of most types of
granulo-monocytic cells. Two widely used myeloid markers, CD13 and CD33,
were not myeloid-specific, because both were clearly positive on
noncommitted progenitor cells. Of 40 antigens tested, CD15 was the only
other marker fulfilling the criteria of a myeloid-specific marker. However,
at concentrations of CD15 that did not induce aggregation, CD15+ cells
constituted less than 50% of the CD34+, CD64+ cells. Furthermore, the
CD34+, CD15- cells showed more than 50% higher CD34 mean fluorescence
intensity than the CD64+, CD15+ cells, indicating that CD64 appears earlier
than CD15 during differentiation. Thus, among a large number of antigens
screened, CD64 was the most useful for the identification and purification
of granulo-monocytic progenitor cells.
Volume 85,
Issue 9,
pp. 2402-2413,
05/01/1995
Copyright © 1995 by The American Society of Hematology
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