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Interleukin-10 increases Bcl-2 expression and survival in primary human
CD34+ hematopoietic progenitor cells
RM Weber-Nordt, R Henschler, E Schott, J Wehinger, D Behringer, R Mertelsmann and J Finke
Department of Hematology & Oncology, Albert-Ludwigs-University Medical
Center, Freiburg, Germany.
Bcl-2 expression has been shown in hematopoietic progenitor cells. Through
the use of Bcl-2 specific antisense oligonucleotides we herein report the
biologic importance of Bcl-2 expression in primary human CD34+
hematopoietic progenitor cells committed to the myeloid lineage. In bone
marrow or peripheral blood derived CD34+ cells Bcl-2 specific antisense
decreased cell survival and inhibited the outgrowth of mixed myeloid
colonies. A short-term overnight pretreatment of CD34+ cells with 25
mumol/L of Bcl-2 antisense in liquid culture completely ablated the growth
of granulocyte-macrophage colony-forming cells (GM-CFC) in a subsequent 14
days methylcellulose colony assay. Control experiments using corresponding
Bcl-2 sense or nonsense oligonucleotides did not significantly impair cell
survival or growth of GM-colony-forming unit. Western blot analyses
revealed the Bcl-2 antisense dependent inhibition of expression of the
Bcl-2 protein in CD34+ progenitor cells. Furthermore, regulation of Bcl-2
expression by various cytokines including interleukin-10 (IL-10) was
studied. IL-10's effects on the formation of mixed myeloid colonies were
examined in the absence or presence of Bcl-2 specific antisense. In the
absence of Bcl-2 antisense IL-10 significantly extended the colony forming
potential of mixed myeloid colonies to 14 days. In the presence of Bcl-2
antisense rhIL-10 completely restored GM-CSF driven colony growth.
Fluorescent microscopy, Western blot analysis, and reverse
transcriptase-polymerase chain reaction revealed the IL-10 dependent
increase in cellular expression of Bcl-2 protein and Bcl-2 mRNA transcripts
in CD34+ cells. Thus these results show that Bcl-2 expression is necessary
for the formation of GM-CSF-dependent colony growth in vitro and that
rhIL-10 increases Bcl-2 expression and survival in primary human CD34+
hematopoietic progenitor cells that are committed to the myeloid lineage.
Volume 88,
Issue 7,
pp. 2549-2558,
10/01/1996
Copyright © 1996 by The American Society of Hematology
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