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Granulomonocyte-associated lysosomal protein expression during in vitro
expansion and differentiation of CD34+ hematopoietic progenitor cells
C Scheinecker, H Strobl, G Fritsch, B Csmarits, O Krieger, O Majdic and W Knapp
Institute of Immunology, Vienna International Research Cooperation Center
at SFI, University of Vienna, Austria.
Using an in vitro expansion and differentiation system for human CD34+ cord
blood (CB) progenitor cells, we analyzed the induction and expression
kinetics of the granulomonocyte associated lysosomal proteins
myeloperoxidase (MPO), lysozyme (LZ), lactoferrin (LF), and macrosialin
(CD68). Freshly isolated CD34+ CB cells were negative for LZ and LF, and
only small proportions expressed MPO (4% +/- 2%) or CD68 (3% +/- 1%).
Culturing of CD34+ cells for 14 days with interleukin (IL)- 1, IL-3, IL-6,
stem cell factor, granulocyte-macrophage colony- stimulating factor
(GM-CSF), and G-CSF resulted in on average a 1,750- fold amplification of
cell number, of which 83% +/- 7% were MPO+. Without addition of GM-CSF and
G-CSF, lower increases in total cell numbers (mean, 211-fold) and lower
proportions of MPO+ cells (54% +/- 11%) were observed. The proportion of
MPO+ cells slightly exceeded but clearly correlated with the proportion of
cells positive for the granulomonocyte associated surface molecules CD11b
(Mac-1), CD15 (LeX), CD64 (Fc gamma RI) CD66, or CD89 (Fc alpha R). At day
14 MPO+ and LZ+ cells were virtually identical. However, at earlier time
points during culture (days 4 and 7), single MPO+ or LZ+ cell populations
were also observed, which only later acquired LZ and MPO, respectively.
Maturation of cells into the neutrophilic pathway was indicated by the
acquisition of MPO, followed by LZ. In contrast, maturation of cells into
the monocytic pathway was indicated by the acquisition of LZ followed by
MPO and CD14. CD68 was found to be expressed at day 4 by the majority of
cells and was not restricted to the granulomonocytic cells, as cells with
megakariocytic (CD41+) or erythroid (CD71hi) features were CD68+. LF
expression was observed only in GM- plus G-CSF- supplemented cultures, in
which only 26% +/- 5% of cells expressed LF by day 14.
Volume 86,
Issue 11,
pp. 4115-4123,
12/01/1995
Copyright © 1995 by The American Society of Hematology
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