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Constitutive expression of steel factor gene by human stromal cells
MC Heinrich, DC Dooley, AC Freed, L Band, ME Hoatlin, WW Keeble, ST Peters, KV Silvey, FS Ey and D Kabat
Department of Medicine, Oregon Health Sciences University, Portland
97201-3098.
Steel factor (SF), the ligand for c-kit, is an essential regulator of
normal hematopoiesis, melanogenesis, gametogenesis, and mast-cell growth
and development. Hematopoietic stromal cells are important sources of SF,
because inactivation of SF in mice results in defects in the support
function of hematopoietic stromal cells. To identify specific cells that
produce, and factors that govern the expression of the different isoforms
of SF in human hematopoiesis, we quantified levels of SF mRNA and
membrane-bound protein in human stromal cells before and after exposure to
recombinant human interleukin (IL)-1 alpha, a cytokine known to induce the
expression of a variety of hematopoietic growth factors. In addition,
because stromal cells in longterm bone marrow cultures (LTBMC) are
supportive of hematopoietic progenitor cell survival in vitro, while
umbilical vein endothelial cells (EC) and diploid fibroblasts (DF) are not,
we also sought to test the hypothesis that SF gene expression would differ
in cells from LTBMC when compared with EC or DF. Using
reverse-transcription polymerase chain reaction amplification (RT-PCR),
ribonuclease protection assays (RPA), and Northern blot analysis, SF was
found to be constitutively transcribed in EC, DF, and LTBMC. IL-1 alpha
neither induced accumulation of SF mRNA nor altered the ratio of exon 6+ to
exon 6- transcripts in these stromal cells. By Northern blot analysis, the
predominant SF mRNA species was shown to be 5.6 kb; a minor population of
3.6 kb was also found. Low levels of membrane-bound SF protein were found
to be constitutively expressed by all three types of stromal cells, and
were not regulated by IL-1 alpha. We conclude that the unique capacity of
LTBMC to support in vitro hematopoiesis, when compared with EC or DF,
cannot be explained on the basis of qualitative or quantitative differences
in SF gene expression in these cells.
Volume 82,
Issue 3,
pp. 771-783,
08/01/1993
Copyright © 1993 by The American Society of Hematology
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