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Differential Effects of Nitric Oxide on Erythroid and Myeloid Colony
Growth From CD34’ Human Bone Marrow Cells
By Paul J. Shami and J. Brice Weinberg
Nitric oxide (NO) is a reactive molecule
with numerous physiologic and pathophysiologicrolesaffecting
the nervous,
cardiovascular, and immune systems. In previous work, we
have demonstrated that NO inhibits the growth
and induces
the monocytic differentiation of cells of the HL-60 cell line.
We have also demonstrated that NO inhibits the growthof
acute nonlymphocytic leukemia cells freshly isolated from
untreated patients and increases monocytic differentiation
antigens in some. In the present work, we studied theeffect
of NO on the growth and differentiation of normal human
bone marrow cells in vitro. Mononuclear cells isolated from
human bone marrow werecultured in semisolid media and
treated with the NO-donating agents sodium nitroprusside
(SNP) or S-nitroso-acetylpenicillamine (SNAP) (0.25 t o 1
mmol/L). Both agents decreased colony-forming unit-erythroid (CFU-E) and colony-forming unit-granulocyte macrophage (CFU-GM) formation by 34% t o 100%. When CD34+
cells were examined, we noted that these cells responded
t o SNP and SNAP differently
than
did the
mono-
nuclear cells. A t a concentration range of 0.25 t o 1 mmol/L,
SNP inhibited the growth
of CFU-E by 30% t o 75%. However,
at thesame concentration range, SNPincreased the number
of CFU-GM by up t o 94%. At concentrations of 0.25 t o 1
mmol/L, SNAP inhibited the growth of CFU-E by 33% t o
100%. At a concentration of 0.25 mmol/L. SNAP did not affect CFU-GM. At higher concentrations, SNAP inhibited the
growth of CFU-GM. Although SNP increased intracellular
levels of cGMP in bone marrow cells, increasing cGMP in
cells byaddition of 8-Br-cGMP (a membrane permeable
cGMP analogue) did notreproduce the observed NO effects
on bone marrow colonies. These results demonstrate that
NO can influence the growth and differentiation of normal
human bone marrow cells. NO (generated in the bone marrow microenvironment) may play an important role modulating the growth and differentiation of bone marrow cells
in vivo.
0 1996 by The American Society of Hematology.
N
were then incubated for 1 hour at room temperature in anAIS
MicroCELLector T-25 flask coated with anti-CD34 antibodies. The
nonadherent cells were discarded, 4 mL of PBS were added, and
the side of the flask was tapped several times to release the CD34’
cells from the antibody coat. The cells were washed in PBS and
resuspended in RPMI-1640 before plating in the clonogenic assays.
The cell fraction obtained was >85% CD34’ by flow cytometry
and constituted 0.8% to 1% of the original cell number. The viability
of the cells after isolation was 100% as determined by Trypan blue
exclusion. In some experiments, the CD34’ population was further
enriched byflow cytometry and sorting. In brief, the CD34’ cell
fraction obtained using the AIS flasks was incubated at 4°C for 30
minutes with fluorescein isothiocyanate-labeled mouse antihuman
CD34’ antibody (Biosource International, Camarillo, CA). The
CD34’ cells were then sorted under sterile conditions using a cell
sorter. The sorted cells were 100% CD34’ and were used for culture
in methylcellulose media as described in the following section.
Culture conditions. Mononuclear bone marrow cells and CD34’
cells were cultured in semisolid media purchased from Stem Cell
Technologies (Vancouver, BC, Canada). The medium contained
30% fetal bovine serum, 10% agar leukocyte conditioned medium,
1% bovine serum albumin, 0.9% methylcellulose, 0.1 m m o K 2mercaptoethanol, 2 mmolL glutamine, and 3 p/mL purified human
urinary erythropoietin. Bone marrow mononuclear cells were cultured at a density of 50,00O/mL. Fifteen thousand cells per well were
plated in 16 mm tissue culture plates. CD34’ cells were cultured at
a density of 10,000 cells/mL. One thousand six hundred cells per
well were plated in 12 mm tissue culture plates. Samples were
cultured in duplicate at37°C in a humidified room air 5% CO,
atmosphere.
ITRIC OXIDE (NO) isa reactive molecule that has
multiple biologic effects. Numerous cells have been
demonstrated to produce NO. These include fibroblasts, endothelial cells, Kupffer cells, mononuclear phagocytes, and
certain neurons.”’ NO has physiologic and pathophysiologic
effects on the cardiovascular, neurologic, and immune systems.’”In previous work, we have demonstrated that NO
inhibits the growth of cells of the HL-60 human myeloid
leukemia cell line.6Along with growth inhibition, we demonstrated that NO induces the monocytic differentiation of HL60 cells and modulates the gene expression of cytokines and
oncogenes6 Wehave also shown that NO inhibits the growth
and induces the differentiation of acute nonlymphocytic leukemia (ANLL) cells freshly isolated from untreated patient^.^
The purpose of the present work was to determine the effects
of NO on the growth and differentiation of normal human
bone marrow cells.Our results show that NO modulates
the growth of erythroid and myeloid colonies. NO could,
therefore, play a significant physiologic and pathophysiologic role modulating the growth and differentiation of bone
marrow cells in vivo.
MATERIALS AND METHODS
Cell source. After obtaining informed consent, iliac crest bone
marrow samples were aspirated from patients undergoing bone marrow cell harvest in preparation for autologous bone marrow transplantation according to a protocol approved by the Duke and VA
Institutional Review Boards. Mononuclear cells were isolated using
a ficoll-Hypaque density gradient and were resuspended in RPMI1640 culture medium (GIBCO, Grand Island, NY). CD34’ cells
were isolated using panning techniques with flasks purchased from
AIS MicroCELLector (Santa Clara, CA) according to the manufacturer’s methods. In brief, 60 X 10‘ bone marrow mononuclear cells
were suspended in 4 mL of phosphate buffered saline solution (PBS)
containing 1 mmoUL ethylene diamine tetra-acetic acid (EDTA)
and 0.5% human gamma globulin. After 15 minutes, the cells were
incubated for 1 hour at room temperature in an AIS MicroCELLector
T-25 flask coated with soybean agglutinin. The nonadherent cells
were collected, washed in PBS and resuspended in 4 mL PBS containing 1 mmolL EDTA and 0.5% human gamma globulin. Cells
Blood, Vol 87, No 3 (February l ) , 1996: pp 977-982
From the Department of Medicine, Division of Hematology-Oncology, VA and Duke University Medical Centers, Durham, NC.
Submitted April 15, 1995; accepted September 12, 1995.
Address reprint requests to J. Brice Weinberg, MD, VA and Duke
University Medical Centers, 508 Fulton St, Durham, NC 27705.
The publicationcosts of this article were defrayed in partby page
charge payment.This article must therefore be hereby marked
“advertisement” in accordance with 18 U.S.C. section I734 solely to
indicate this fact.
0 1996 by The American Society of Hematology.
0006-4971/96/8703-0013$3.00/0
977
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978
SHAM1 AND WEINBERG
1251+
cGMP rneasurernenfs. Mononuclear cells isolated from the bone
marrow by density centrifugation were cultured at a density of 1 x
CFU-E
CFU-GM
10‘ cells/mL in RPMI-I640 at37°C in a humidified atmosphere with
5% COz. The cells were harvested at 15, 30, 45, and 60 minutes,
100
lysed in ethanol, and cGMP levels were measured using the Scintillation Proximity Assay (Amersham, Arlington Heights,1L) according
to the manufacturer’s recommendations. Briefly, cells were lysed in
75
2,OOOg at4”C,the
65%icecoldethanol.Aftercentrifugationat
supernatant was collected and evaporated at60°C in a vacuum oven.
Cell extracts were then suspended in assay buffer supplied by the
50
manufacturer. One hundred microliters of cell extract were mixed
with 100 pL of 1-121 labeled cGMP solution. One hundred microli25
ters of rabbit anti-cGMP antiserum were added followed by 100 p L
of donkey antirabbit IgG antibody coupled to fluomicrospheres (all
solutions and reagents supplied by the manufacturer). The suspension was incubatedatroomtemperaturefor
15 to 20 hours.The
samples’luminescencesweremeasured
in ascintillationcounter.
Measurementsweremade
in duplicate.ThecGMPconcentration
-25
was deduced from a standard curve and expressed
as picograms/
million cells.
0.75
0.25
0.5
1
0
NO donors and chemicals. Sodium nitroprusside (SNP), and Snitrosoacetyl penicillamine (SNAP) were used as NO sources. SNP
SNP (mM)
was purchased from Elkins-Sinn (Cherry Hill, NJ), and SNAP was
purchased from Chem Biochem Research Inc (Salt Lake City, UT).
Fig 1. Effect of SNP on the growth of colonies from the mononuSNP and SNAP solutions were made immediately before use and
clear fraction of the bone marrow. SNP was added directly to the
added directly to the cell cultures at the indicated
final concentrasemisolid methylcellulosecell suspension at theindicated concentrations. The cultures that were treated with SNP were exposed to light tions. Colonies were scored at days 10 to 18. There was an average
of 52 CFU-E and 14 CFU-GM per well in the controls. SNP inhibited
for 30 seconds to enhance NO release by a photolytic effect. It is
the growth of CFU-E and CFU-GM in a dose-dependent fashion. The
NO donors do not
important to note that the concentrations of the
data points represent means 2 one standard error of the mean. Astercorrespond to the amount of NO delivered to the bone marrow cells
isks indicate statistically significant differences (P .05) between
in solution. The actual NO concentration delivered to the cells
is
treatments and controls. Results shown are the averages of three
likely much less because of the very short half-lifeof NO in solution.
separate experiments.
8-Bromoguanosine3’,5’-cyclicmonophosphate(8-Br-cGMP)was
(St Louis, MO). 8-Brpurchased from Sigma Chemical Company
cGMP solutions were made immediately before
use and added to
Effect of SNAP on the mononuclearfraction of bone marthe cell cultures at a final concentration of 3 mmol/L.
row cells. To ascertain that the growth inhibitory effects
Analysis. Colonies were scored IO to 18 days after plating using
observed with SNP treatment were due to NO and not to the
an inverted light microscope. A colony was defined as a group of
NO-carrying molecule, we treated cells obtained from the
30 or more cells. Under our culture conditions, the majority of the
mononuclear fraction of the bone marrow withSNAP. SNAP
erythroidcoloniesobservedatthetime
of scoringwerecolonyis a compound that liberates NO in solution.KBone marrow
forming unit-erythroid (CFU-E) colonies and did not display a typicells were cultured in semisolid media with increasing concal burst configuration. Therefore, we used only these colonies to
analyze the effect ofNO on erythroid precursors. Statistical analysis centrations of SNAP as described in the Materials and Methwasdone using theStudent’st-test.Differenceswereconsidered
ods section. Colonies were assayed at days 10to 18. At
statistically significant for P < .05.
concentrations of 0.25 to 1 mmoVL, SNAP inhibited the
-
-
O4
i
*
*
-=
RESULTS
Effect of SNP on the mononuclearfraction of bone murrow cells. We first studied the effect of SNP on colony
formation using the bone marrow mononuclear fraction as
a source of progenitor cells. SNP did not affect the viability
of the bone marrow cells when tested by trypan blue exclusion. SNP was added to bone marrow cells cultured in semisolid media as described in Materials and Methods. Colonies
were assayed at days 10 to 18. At concentrations of 0.25 to
I mmol/L, SNP inhibited the growth of CFU-E by 42% to
80% (with P < .OS for each treatment), and CFU-GM by
34% to85% ( P < .OS for the 0.5, 0.75, and 1 mmoVL
treatments) (Fig 1). The colonies obtained from cells treated
with SNP were smaller than the colonies from untreated
cells. However, this effect was difficult to quantify. There
was no significant difference between the extent of CFU-E
inhibition and CFU-GM inhibition at all the SNP concentrations used.
growth of CFU-E by 66% to 100% ( P < .OS for all treatments) (Fig 2). At the same concentrations, it inhibited the
growth of C m - G M by 53% to 100% ( P < .OS for treatments
SNP
of 0.5, 0.75, and 1 mmol/L) (Fig 2). Asnotedwith
treatments, the colonies obtained from cells treatedwith
SNAP were smaller than colonies obtained from untreated
cells. There was no significant difference between the extent
of CFU-E inhibition and CFU-GM inhibition at all the SNAP
concentrations used.
Effectof SNP on CD34+ cells. The growth inhibitory
effects of NO described above could be due to one or more
of the following effects: a direct effect on bone marrow
progenitors, induction of growth inhibitory factors in accessory cells, and inactivation of growth factors or their receptors. To determine whether NO acted directly on progenitor
cells, we enriched the progenitor cell population by positively selecting CD34’ cells from the bone marrow using a
panning technique with anti-CD34 antibody. CD34+ enriched cells constitute a population of enriched progenitors
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979
NITRIC OXIDE AND HUMAN CD34'CELLS
'251100
CFU-GM
CFU-E
-
75 50
-
25 -
0-
*
*
I
1
I
I
0
0.25
0.5
0.75
*
I
1
SNAP (mM)
Fig 2. Effect of SNAP on the growth colonies
of
from the mononuclear fraction of the bone marrow.SNAP was added directly to the
semisolid methylcellulose cell
suspension at theindicated concentrations. Colonies were scored at days 10 t o 18. There was an average
of 32 CFU-E and 22 CFU-GM per well in the controls. SNAP inhibited
the growthof CFU-E and CFU-GM in a dose-dependent fashion. The
data pointsrepresent means f one standard errorof the mean. Asterisks indicate statistically significant differences (P< .05) between
treatments and controls. Results shown are the averages of two
separate experiments.
with very low numbers of mature or accessory cells. CD34+
cells were cultured in semisolid media in the presence of
increasing concentrations of SNP. Colonies were scored at
days 10 to 18. At a concentration of 0.25 to 1 mmol/L, SNP
decreased the number of CFU-E obtained from CD34+ cells
by 30% to 75% (with P < .05 for all the treatments) (Fig
3). However, at the same concentrations, it increased the
growth of CFU-GM by up to 94% ( P < .05 for the 0.5,
0.75, and 1 mmol/L treatments) (Fig 3). To confirm these
results, we repeated the experiments using a population of
100% CD34' cells obtained by cell sorting as described in
the Materials and Methods section. When such cells were
used, SNP (0.25 to 1 mmol/L) inhibited the growth of CFUE by 29% to 95%, but did not affect the growth of CFUGM (data not shown).
Effect of SNAP on CD34' cells. To provide further evidence that the observed effects on CD34+ cells were due to
NO, we did experiments in which CD34+-enriched cells
were cultured in semisolid media and treated with increasing
concentrations of SNAP. At concentrations of 0.25 and 0.5
mmol/L, SNAP inhibited the growth of CFU-E by 33% and
81%, respectively ( P < .05 for both treatments) (Fig 4). At
a concentration of 0.25 mmol/L, it did not inhibit Cm-GM,
but at a concentration of 0.5 mmoyL it inhibited CFU-GM
by 30% (Fig 4). However, the latter change did not reach
statistical significance. At concentrations of 0.75 and 1
mmol/L, SNAP totally inhibited the growth of CFU-E and
CFU-GM ( P < .05) (Fig 4).
Effect of SNP on cGMP levels in bone marrow cells. One
of the main intracellular targets of NO is the soluble guanylate cyclase
For example, NO induces the relaxation of vascular smooth muscle cells by increasing intracellular levels of cGMP through the activation of guanylate
cyclase.' Consequently, we determined the effect of NO on
cGMP levels in bone marrow cells. Mononuclear cells from
the bone marrow were isolated by density centrifugation and
cultured in RPMI-1640 at a density of 1 X lo6 cells/mL.
The cells were treated with 1 mmoln SNP and harvested
after 15, 30, 45, and 60 minutes. SNP increased the intracellular levels of cGMP in a time dependent manner with almost
a sixfold increase at 60 minutes as compared with controls
(the differences reached statistical significance with P < .05
for the 30-, 45-, and 60-minute time points) (Fig 5).
Effect of cGMP on bone marrow and CD34' cells. Since
NO increased intracellular levels of cGMP in bone marrow
cells, we sought to determine whether cGMP would reproduce the NO-induced changes in the bonemarrow.We,
therefore, treated mononuclear cells obtained from the bone
marrow with the membrane permeable cGMP analog 8-BrcGMP. When added to bone marrow mononuclear cells at
a concentration of 3 mmoVL, 8-Br-cGMP had no effect on
CFU-E, but it inhibited CFU-GM by 53% ( P < .05) (Fig
6A). When added to CD34+-enriched cells at the same concentrations, 8-Br-cGMP had no effect on CFU-E, but it inhibited CFU-GM by 34% ( P < .05) (Fig 6B).
DISCUSSION
In the present work, we have used clonogenic assays to
study the effect of NO on human bone marrow
growth. When
300 CFU-GM
CFU-E
n
m
2
2
8
250-
rr
O
200-
c)
E
Q)
*
2 150-
Q)
a
W
.CI 100E
0
I
6
50-
I
I
I
I
0
0.25
0.5
0.75
1
1
SNP (mM)
Fig 3. Effect of SNP on the growthof colonies from CD34+ cells.
SNP was added directly t o the semisolid methylcellulose cell
suspension at the indicatedconcentrations. Colonies were scored at days
10 t o 18. There was an average of 98 CFU-E and 33 CFU-GMper well
in the controls. SNP inhibited the growth CFU-E
of
and enhanced the
growth ofCFU-GM. The data pointsrepresent means f one standard
error of the mean. Asterisks indicate statistically significant differbetween treatments andcontrols. Results shown are
ences (P< .M)
the averages of four separate experiments.
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SHAM1 AND WEINBERG
CFU-E
125 -
"f-
CFU-GM
T
75 -
100
50 25 -
0-
*
*
-25 I
0.75
I
00.5
I
0.25
I
*
1
1
SNAP (mM)
Fig 4. Effect of SNAP on the growth of colonies from CD34' cells.
SNAP was added directly to the semisolid methylcellulose cell suspension at the indicated concentrations.Colonies were scored at
days 10 to 18. There was an average of 82 CFU-E and 34 CFU-GM
per well in the controls. SNAP inhibited the growth of CFU-E at all
the concentrations used. SNAPdid not affect CFU-GM at 0.25 mmoll
L but inhibited their growth at higher concentrations.The data points
represent means f one standarderror ofthe mean. Asterisks indicate
statisticallysignificant differences( P < .05) between treatments and
controls. Results shown are the averages of three separate experiments.
the whole mononuclear fraction of the bone marrow was
used as a source of progenitor cells, NO inhibited the growth
of both C W - E and CFU-GM. NO increased intracellular
levels of cGMP in these cells, and exogenously added 8-BrcGMP partially reproduced the NO effects. However, when
the progenitor cell population was enriched and treated with
NO, the erythroid and myeloid lineages were affected differently. SNP inhibited erythroid colony growth, but enhanced
myeloid colony growth. When SNAP was used as a source
of NO, erythroid colonies were inhibited at concentrations
of 0.25 and 0.5 mmoVL, while myeloid colony growth was
not significantly inhibited at the same concentrations. At
higher concentrations, SNAP inhibited both erythroid and
myeloid colony growth totally. While exogenously added 8Br-cGMP partially reproduced the NO effects on the mononuclear fraction of the bone marrow, it did not have the same
effects on CD34' cells.
Our results suggest that NO may affect the growth of
bone marrow colonies from precursor cells both directly and
indirectly. In the presence of accessory and mature mononuclear cells, it inhibits the growth of both C W - E and CFUGM. It could, therefore, induce the production of cytokines
or factors in the accessory cells, which in turn, would inhibit
progenitor cell growth. In fact, we have previously reported
that NO increases the mRNA levels of tumor necrosis factora and interleukin l-p in cells of the IU-60 human myeloid
leukemia cell line.6 NO has also been shown to enhance
the production of tumor necrosis factor in interleukin-l-a
stimulated human mononuclear cells." A similar effect could
be operating here. On the other hand, NO had different effects when added to CD34+ cells. When added to this population of enriched progenitor cells, it inhibited the growth of
CFU-E, but enhanced the growth of CFU-GM. This growth
enhancing effect was seen when SNP was used as a source
of NO. When SNAP was used at 0.25 mmol/L, ithadno
effect on CFU-GM, even though it inhibited CFU-E.
The discrepancies between the observed effects of SNP
and SNAP on CD34+ cells are most likely due to the different amounts of NO liberated in solution by the two compounds. At all the concentrations used, SNAP had a more
marked growth inhibitory effect than did SNP. The effect of
NO on bonemarrow growth and differentiation therefore
appear tobe concentration-dependent. At high concentrations, it potently inhibits the growth of both the erythroid
and myeloid pathways. At lower concentrations, it seems to
act as a signal that preferentially inhibits the growth of erythroid cells and enhances the growth of myeloid cells. The
latter effect may be due to a direct effect on the bone marrow
progenitors because it was observed only when an enriched
population of CD34+ cells was used as a cell source. Even
though it increased the intracellular levels of cGMP in bone
marrow cells, the NO effects appear to be cGMP-independent because raising intracellular cGMP levels with the
membrane permeable cGMP analogue 8-Br-cGMP did not
reproduce the NO effects.
NO has been reported to affect cell growth in different
settings. It inhibits tumorcell growth andis cytostatic or
*
6-
4-
2-
T
0
15
30
45
60
Minutes
Fig 5. Effect of SNP on cGMP levels in bone marrow mononuclear
cells. SNPwas addad to bone marrow mononuclear cdle cultured in
RPMI-1640. The cells were lysed at the indicated time points and
intracellularcGMP levels were measured. The data points represent
means 2 one standard error of the mean. Astarisks indicate statistically significant differences ( P < . 0 5 ) between treatments and controls. Results shown are the averages of two separate experiments.
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98 1
NITRICOXIDE AND HUMAN CD34' CELLS
Fig 6. Effect of 8-Br-cGMPon the growthof bone
marrow colonies. 8-Br-&MP was added directly to
the semisolid methylcellulose cellsuspension at a
concentrationof 3 mmol/L. Colonies were scored at
days 10 to 18.8-Br-&MP did not affect the growth
of CFU-E, but inhibked the growth of CFU-GM. The
figures show resultsof experimentsusing the mononuclear fraction of the bone marrow (A) or CD34+
(B) cells as a source of progenitors.The data points
represent means one standard error of the mean.
Asterisks indicate statisticallysignificant differences
( P < .W)
between treatments and controls. Results
shown are the averages oftwo separateexperiments
for each figure.
*
cytocidal for certain microorganisms.'.' NO has also been
shown to inhibit lymphocyte proliferation." NO also affects
cellular progression through the cell cycle of murine macrophage-like cells." While Pipili-Synetos et a l l 3 demonstrated
that NO could inhibit angiogenesis, Ziche et all4 have shown
that NO mediates angiogenesis and promotes endothelial cell
growth and migration. NO has also been shown to amplify
fibroblast growth factor-2-induced mitogenesis of rat aortic
smooth muscle cell^.'^ Punjabi et all6 have shown that interferon-y and lipopolysaccharide inhibited murine bone marrow colony growth through a NO-dependent mechanism.
Finally, Maciejewski et a l l 7 reported that NO inhibits the
growth of human bone marrow cells in vitro. They did not
observe a selective effect on either the erythroid or the myeloid lineage.
NO can alter cell growth and differentiation by affecting
different intracellular targets. Although the guanylate cyclase-cGMP signal transduction pathway is a major target
of NO in the vascular sy~tem,~*'*~
it is not clear from the
results we report here that NO affects the hematopoietic
system solely through cGMP. Other potential mechanisms
of action and intracellular targets include protein sulfhydryl
group nitrosylation,' mitochondrial respiration," G protein
activation," activation of nuclear factor-KB,20inhibition of
ribonucleotide reductase,"." protein adenosine diphosphate
r i b ~ s y l a t i o n and
, ~ ~DNA
~ ~ ~ damage and mutation.** NO has
also been shown to induce apoptosis in murine peritoneal
macrophages and in human bone marrow ~ e l l s . ' ~It, ' is
~ also
remarkable that NO atverylow
concentrations prevents
apoptosis in B cells through the maintenance of bcl-2 leve1s.27
It is clear from our observations and from those reported
by other investigators that NO can play multiple physiologic
and pathophysiologic roles affecting bone marrow cell
growth and differentiation. It affects bone marrow physiology at different levels. It can mediate increased blood flow
to the marrow under hematopoietic stress." Our work and
the work of
show that it can also have a significant
effect on the growth of cells of the erythroid and myeloid
pathways. This is especially significant because the bone
marrow contains numerous cells that could be potential
sources of NO. At low to moderate levels, NO could selectively switch bone marrow cell production from the erythroid
to the myeloid pathway. At high levels of production, it
could potently inhibit the growth of both erythroid and my-
Variable
Varlable
A
B
eloid cells. NO could, therefore, be an important mediator
in the generation of the anemia associated with chronic inflammatory states (such as rheumatoid arthritis and cancer).
Therapeutic modulation of NO production andor NO effects
in vivo might significantly affect hematopoiesis.
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982
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From www.bloodjournal.org by guest on July 28, 2017. For personal use only.
1996 87: 977-982
Differential effects of nitric oxide on erythroid and myeloid colony
growth from CD34+ human bone marrow cells
PJ Shami and JB Weinberg
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