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Prepublished online as a Blood First Edition Paper on May 24, 2002; DOI 10.1182/blood-2002-01-0062.
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Blood, 15 August 2002, Vol. 100, No. 4, pp. 1318-1325
HEMATOPOIESIS
Functional interleukin-7 receptors (IL-7Rs) are expressed by
marrow stromal cells: binding of IL-7 increases levels of IL-6 mRNA and
secreted protein
Mineo Iwata,
Lynn Graf,
Norihiro Awaya, and
Beverly Torok-Storb
From the Clinical Research Division, Fred Hutchinson
Cancer Research Center, Seattle, WA.
DNA spotted microarrays were used to compare gene expression
profiles from 2 functionally distinct human marrow stromal cell lines:
HS-27a, which supports cobblestone area formation by early hematopoietic progenitors, and HS-5, which secretes multiple cytokines that support the proliferation of committed progenitors. One unexpected result was the high level of interleukin-7 receptor (IL-7R) gene expression in HS-27a stromal cells. Northern blot analysis confirmed the IL-7R RNA expression, and Western blots for the IL-7R protein detected both a full-length (90-kd) IL-7R and a smaller 30-kd fragment
in both HS-27a cells and primary stromal cell cultures, whereas only
the 90-kd receptor protein was detected in peripheral blood mononuclear
cells. Biotinylated IL-7 was shown to bind to HS-27a cells
under physiologic conditions, and this binding was inhibited by
blocking anti-IL-7 antibodies. Tyrosine phosphorylation of several
proteins (55 kd, 30 kd, and 24 kd) in HS-27a cells was rapidly
increased after incubation with recombinant IL-7. One of the
phosphorylated proteins proved to be the 30-kd IL-7R fragment. Exposure
of HS-27a cells to IL-7 resulted in a 10-fold increase in secretion of
IL-6 into culture supernatants but no increase in the cytokines stromal
cell-derived factor 1, macrophage inflammatory protein 1 ,
or IL-1 . The up-regulation of IL-6 secretion is associated
with a rapid but transient increase in detectable levels of IL-6
messenger RNA. These data suggest that IL-7 may function to regulate
the milieu of the microenvironment by modulating IL-6 secretion by the
IL-7R-expressing stromal elements.
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