|
|
 Previous Article | Table of Contents | Next Article 
Blood, Vol. 93 No. 5 (March 1), 1999:
pp. 1579-1585
Receptor Clearance Obscures the Magnitude of Granulocyte-Macrophage
Colony-Stimulating Factor Responses in Mice to Endotoxin or Local
Infections
By
Donald Metcalf,
Nicos A. Nicola,
Sandra Mifsud, and
Ladina Di Rago
From the Division of Cancer and Haematology, The Walter and Eliza
Hall Institute of Medical Research, Royal Melbourne Hospital, Victoria,
Australia.
 |
ABSTRACT |
Marrow cells from mice lacking high-affinity receptors for
granulocyte-macrophage colony-stimulating factor (GM-CSF;
 c / mice) were shown to bind and internalize much
less GM-CSF than cells from normal (c+/+) mice.
c/ mice were used to determine the effect of
negligible receptor-mediated clearance on detectible GM-CSF responses
to the intravenous injection of endotoxin or the intraperitoneal
injection of casein plus microorganisms. Unlike the minor serum GM-CSF
responses to endotoxin seen in c+/+ mice, serum
GM-CSF levels rose 30-fold to 9 ng/mL in c/ mice
even though loss of GM-CSF in the urine was greater than in
c+/+ mice. Organs from c/ and
c+/+ mice had a similar capacity to produce GM-CSF
in vitro, as did peritoneal cells from both types of mice when
challenged in vitro by casein. However, when casein was injected
intraperitoneally, c/ mice developed higher and
more sustained levels of GM-CSF than did c+/+ mice.
The data indicated that receptor-dependent removal of GM-CSF masks the
magnitude of GM-CSF responses to endotoxin and local infections.
Because of this phenomenon, serum GM-CSF concentrations can be a
misleading index of the occurrence or nonoccurrence of GM-CSF responses
to infections.
© 1999 by The American Society of Hematology.
| |
INTRODUCTION |
THE GLYCOPROTEIN COLONY-stimulating
factors were first detected because of their proliferative action in
vitro on hematopoietic cells.1 Subsequent studies have
shown that these regulators have a variety of additional actions,
including stimulation of the functional activity of mature cells in
responding lineages.2
Serum and tissue concentrations of colony-stimulating factors are low
under basal conditions3 but are elevated in infections or
in response to the injection of bacterial lipopolysaccharide (LPS).2,4,5 The presumption is that these responses are designed to elicit prompt antimicrobial responses by activating existing mature cells and, if necessary, to sustain and amplify such
responses by stimulating the production of additional cells.
Gene inactivation studies have indicated that granulocyte
colony-stimulating factor (G-CSF) is a major regulator of granulocyte production.6 Despite its stronger proliferative action in
vitro, loss of granulocyte-macrophage colony-stimulating factor
(GM-CSF) or its high-affinity receptor has no apparent effect on the
numbers of granulocytes, monocytes, or eosinophils.7,8
However, mice lacking GM-CSF develop alveolar proteinosis often with
secondary lung infections, diseases that are likely to be based on loss of an essential stimulating action of GM-CSF on macrophage function. Mice with inactivation of the genes encoding G-CSF and/or
GM-CSF die prematurely with a miscellany of infections.9
Although serum levels of G-CSF can be high in spontaneous and
experimental infections, serum GM-CSF levels are typically low or
undetectible in these situations.10-13 This pattern has led many to conclude that GM-CSF is likely to be of only minor importance as a mediator of responses to microbial infections.
As with other glycoprotein regulators, the serum half-life of agents
such as thrombopoietin or GM-CSF is strongly influenced by
receptor-mediated endocytosis and degradation of the regulator by
responding cells.14,15 In principle, observed serum levels of a regulator may be low as a consequence of this process and then be
a misleading index in monitoring regulator responses elicited by an
inducing stimulus.
In the present studies, mice lacking high-affinity receptors for GM-CSF
because of inactivation of the gene encoding the -common chain of
the GM-CSF receptor (c/ mice)8
were used to reexamine GM-CSF responses to the injection of LPS and to
a simple model bacterial infection. Changes in G-CSF levels were
monitored as a control for the general responsiveness of the mice. The
data revealed that c/ mice do respond to
LPS by sharply elevating serum GM-CSF levels and to local infections by
clear elevations of local GM-CSF concentrations, suggesting that these
responses are largely obscured in normal animals because of
receptor-mediated clearance by cells that include the hematopoietic
cells responding to GM-CSF.
| |
MATERIALS AND METHODS |
Mice.
The generation of C57BL6x129Sv mice lacking high-affinity
-common chain (c) receptors for GM-CSF has been
described previously.8 This knockout line is maintained by
interbreeding of mice with homozygous inactivation of the gene encoding
the -common chain (c/ mice). Control
(c+/+) mice used were C57BL6x129Sv mice interbred in
parallel. All mice were raised and housed under pathogen-free
conditions and were used when aged between 2 and 3 months.
Binding and internalization studies.
125I-labeling of recombinant murine GM-CSF (Pepro Tech,
Rocky Hill, NJ) was performed as described previously.16
Internalization and degradation of 125I-murine GM-CSF by
c/ and control bone marrow cells were
performed and analyzed using an acid elution method validated by
extensive previous studies and described in full
previously.16,17 Briefly, bone marrow cells were
resuspended in RPMI medium containing 10% (vol/vol) fetal calf serum
(FCS) and 10 mmol/L HEPES buffer pH 7.4 at 8.5 × 107
cells/mL and equilibrated at 37°C. At time zero,
125I-GM-CSF was added at 100 ng/mL (5 × 106 cpm/mL) with or without unlabeled GM-CSF at 2 µg/mL.
At indicated timepoints 100 µL aliquots were removed (in duplicate),
the cells centrifuged through a cushion of FCS, and resuspended in 1 mL of 3% acetic acid in phosphate-buffered (10 mmol/L, pH 7.4) saline (0.15 mol/L NaCl). The cells were centrifuged, and the radioactivity eluted by acetic acid (cell surface) or retained within the cell (internal) was measured in a gamma counter. Specific counts were determined in each case by subtracting counts present in equivalent incubations in which unlabeled GM-CSF was present.
Endotoxin.
Endotoxin (lipopolysaccharide, Difco, Detroit, MI) was dissolved in
0.9% sodium chloride solution, and 5 µg was injected intravenously in an injection volume of 0.2 mL. Urine was collected at intervals after endotoxin injection and low-molecular-weight inhibitors removed
by passage through an NAP-5 column (Pharmacia, Uppsala, Sweden). At
intervals after injection, mice were anesthetized using penthrane and
bled from the axilla to collect serum.
Local peritoneal cavity inflammation and infection.
Mice were injected interperitoneally with 2 mL of an 0.2% (wt/vol)
solution of casein (Glaxo Laboratories, Melbourne, Australia) or casein
C5890 (Sigma Chemical Co, St Louis, MO) in mouse tonicity phosphate-buffered saline (MTPBS). The Glaxo preparation contains a
contaminating population of nonviable bacteria, and the Sigma preparation is contaminated by viable saphrophytic Bacillus
organisms.18 In both cases, the organisms are cleared by
local neutrophil phagocytosis during the following 2 hours.18 At intervals up to 3 hours after injection, the
mice were anesthetized then their blood collected from the axilla. The
abdominal cavity was then injected with 2 mL MTPBS and, after massage
to mix peritoneal cells with injected harvesting fluid, the cell
suspension was removed using a soft plastic pipette. The collected
volume was usually 3 mL.
Production of peritoneal cell conditioned media.
Resident peritoneal cells were collected using MTPBS and, after
washing, 1 × 106 cells were incubated at 37°C for
3 hours in 1 mL Dulbecco's modified Eagle's medium (DMEM) containing
10% newborn calf serum and 0.1 mL MTPBS or 0.1 mL of a 2% solution of
casein in MTPBS. Media were then obtained, cells removed by
centrifugation then millipore filtration, and the media stored at
4°C before assay.
Production of organ-conditioned media.
Minced organs from 2- to 3-month-old female
c/ and c+/+ mice were
incubated for 4 days in 1-mL volumes of serum-free DMEM as described
previously.19 The media were obtained then millipore filtered and stored at 4°C before assay for CSF content.
Agar cultures.
Agar cultures were performed in 1-mL volumes containing 50,000 bone
marrow cells from C57BL6x129Sv mice in 35-mm plastic petri dishes.1 The medium used was DMEM containing a final
concentration of 20% newborn calf serum and 0.3% agar. Colony
formation was stimulated in replicate cultures by addition of serial
dilutions of 0.1 mL of recombinant murine GM-CSF or human G-CSF.
Cultures were incubated for 7 days in a fully humidified atmosphere of 10% CO2 in air. After initial scoring at
×35, cultures were fixed by the addition of 1 mL of
2.5% glutaraldehyde. Four hours later, the cultures were floated
intact onto glass slides and, after drying, were stained in sequence
for acetylcholinesterase, then with Luxol Fast Blue (BDH
Laboratory, Poole, UK) and hematoxylin.1 After mounting
under coverslips, the cultures were analyzed at ×200 and ×100 to
determine the number and composition of colonies in the entire cultures.
GM-CSF and G-CSFassays.
Concentrations of GM-CSF in serum, urine, peritoneal fluid, and cell-
or organ-conditioned media were determined by microwell assays using
Lux 60 well microtiter trays (Nunc, Naperville, IL) containing 200 FDC-P1 cells in 100 µL DMEM with 10% FCS.19 Serial twofold dilutions of 5 µL of the test material were added to
duplicate wells and, after incubation for 48 hours at 37°C in a
fully humidifed atmosphere of 10% CO2 in air, viable cells
were counted using an inverted microscope. GM-CSF concentrations were
calibrated using a parallel titration of 1 ng/ mL of purified
recombinant murine GM-CSF (Pepro Tech).
FDC-P1 cells respond to stimulation either by GM-CSF or
interleukin-3 (IL-3). The validity of the present assays as specific measurements of GM-CSF was verified by showing that all FDC-P1 stimulating activity in the samples tested was able to be inhibited by
1 µg/mL of a rat anti-mouse GM-CSF monoclonal antibody (Genzyme, Cambridge, MA), an antibody having no inhibitory action on the stimulation of FDC-P1 cells by IL-3. All serum samples were also assayed in parallel using Ba/F3 cells, a cell line responding only to
stimulation by IL-3. No stimulation was observed with any sample,
indicating the absence of detectible IL-3.
The assay of serum and urine concentrations of G-CSF was performed
using a similar general procedure but using 200 Ba/F3 cells engineered
to express G-CSF receptors.20 G-CSF concentrations were
calibrated using parallel titrations of purified recombinant human
G-CSF (Amgen, Thousand Oaks, CA).
The lower detection limit of both CSF assays was 50 pg/mL. Titrations
of serum GM-CSF and G-CSF were commenced using a 1:4 serum dilution to
avoid the inhibiting effects of serum lipoprotein, and the lower
detection limits for sera were therefore 200 pg/mL.
| |
RESULTS |
Colony formation stimulated by GM-CSF or G-CSF.
Titration of recombinant GM-CSF or G-CSF using cultures of 50,000 control C57BL6x129Sv (c+/+) marrow cells showed
(Fig 1) that maximal numbers of
granulocytic and/or macrophage colonies were stimulated to
develop by concentrations of 5 to 10 ng/mL of either CSF. Use of higher
G-CSF concentrations, up to 1 µg/mL, did not further increase the
number of colonies developing and had only a moderate effect on mean
colony cell numbers, which remained relatively low.
View larger version (13K):
[in this window]
[in a new window]
| Fig 1.
Stimulation of colony formation by recombinant GM-CSF or
G-CSF in agar cultures of 50,000 control C57BL6x129Sv bone marrow
cells. Each point represents the mean value from duplicate cultures.
|
|
Internalization of GM-CSF by
c/ and
c+/+ cells.
The ability of c/ or c+/+
bone marrow cells to use up GM-CSF was evaluated by incubating cells
with 125I-labeled murine GM-CSF (100 ng/mL) at 37°C and
measuring cell surface-associated (acid elutable) and internalized
(nonelutable by acid) radioactivity (Fig
2). As expected, given the presence of only low-affinity receptors on
c/ bone marrow cells,8
significantly less GM-CSF was bound by c/
cells (~2,000cpm) than bound by an equivalent number of
c+/+ cells (~14,000cpm). In addition, GM-CSF was
rapidly internalized in c+/+ cells (ke = 0.12/min) and after 30 minutes showed clear evidence of degradation of
GM-CSF because total cell-associated counts decreased to about 3,000 cpm (80% degradation by 3 hours). In contrast, very little if any of
the cell-associated GM-CSF was internalized by
c/ cells, and this situation did not
change with time. This very weak internalization precluded any accurate
estimate of internalization or degradation rates but allowed the
conclusion that little internalization occurred.
View larger version (17K):
[in this window]
[in a new window]
| Fig 2.
Binding and internalization of 125I-labeled
rmGM-CSF to c+/+ bone marrow cells (upper panel) and
the much lower binding and insignificant internalization by
c/ marrow cells (lower panel).
|
|
Serum GM-CSF and G-CSF responses to intravenous endotoxin.
In agreement with previous studies, the intravenous injection of 5 µg
endotoxin induced major rises in serum CSF levels peaking 3 to 6 hours
after injection.19,21 In control c+/+ mice,
G-CSF was not detectible in the serum of uninjected mice but became
detectible 1 hour after injection, then rose sharply and attained a
peak at 6 hours of 1,000 ng/mL (Fig 3).
Serum G-CSF levels were still elevated at 24 hours after injection.
GM-CSF also was not detectible in the serum of uninjected
c+/+ mice and, in agreement with previous
studies,21 barely detectible levels of GM-CSF were present
in serum from 1 to 3 hours after endotoxin injection, thereafter again
becoming undetectible.
View larger version (24K):
[in this window]
[in a new window]
| Fig 3.
Serum concentrations of GM-CSF and G-CSF after the
intravenous injection of 5 µg endotoxin to c+/+ or
c/ mice. Each point represents mean CSF values ± SD from three separate mice of each genotype at each timepoint.
|
|
c/ mice injected with 5 ug endotoxin
showed identical G-CSF responses to those seen in c+/+
mice. In the serum of uninjected c/ mice,
GM-CSF was present, but in barely detectible concentrations. However,
in sharp contrast to the situation with control mice, GM-CSF was
readily detectible in the serum of c/ mice
1 hour after the injection of endotoxin, with concentrations rising
gradually to a peak of 9 ng/mL 3 hours after injection. After 6 hours,
GM-CSF concentrations declined and again were barely detectible at 24 hours.
Although serum GM-CSF levels have been noted to be higher in male than
in female c/xGM-CSF transgenic mice,15
it was of interest that after endotoxin injection, GM-CSF
concentrations in female c/ mice were 2- to 16-fold higher than in corresponding male
c/ mice.
Validation of the bioassays.
Bioassays were used in preference to immunoassays because they
certified that the material being assayed was biologically active.
Previous studies on c/ mice showed a major
difference between the half-life of injected GM-CSF when comparing
bioassays with half-lives established using radiolabeled GM-CSF, the
difference being due to inactivation of much of the GM-CSF, although it
remained macromolecular.8,15
The FDC-P1 cells used in the bioassays also respond to proliferative
stimulation by IL-3. Previous studies showed that no detectible IL-3
appears in mouse serum after the injection of endotoxin.19,21 That the active molecule being assayed in
the FDC-P1 assays was indeed GM-CSF was verified in the present
experiments by showing that all FDC-P1-stimulating activity in the
sera was neutralized by a monoclonal GM-CSF antibody as shown in the
examples in Fig 4. This antibody had no
effect in parallel FDC-P1 cultures stimulated by IL-3.
View larger version (18K):
[in this window]
[in a new window]
| Fig 4.
(Upper panel) Stimulation of the proliferation of FDC-P1
cells by serial dilutions of 1 ng/mL GM-CSF or IL-3 and the selective
inhibition of GM-CSF but not IL-3 by a monoclonal  GM-CSF antibody.
(Lower panel) The stimulating activity of postendotoxin sera for FDC-P1
cells is completely inhibited by the same GM-CSF antibody.
|
|
The presence of IL-3 in the serum samples was further excluded by
assaying all sera in parallel in cultures of Ba/F3 cells, a cell line
responding only to stimulation by IL-3.20 No stimulating activity was detected in any sample. This permitted the use of Ba/F3
cells, engineered to express receptors for G-CSF, to be used as a
specific assay for G-CSF.
Urine GM-CSF and G-CSF levels after intravenous endotoxin.
Previous studies have shown that intravenously injected GM-CSF appears
in the urine, although less than 1% of injected GM-CSF is cleared from
the body by this route.15,22 It was also shown that more
injected GM-CSF appeared in the urine of
c/ mice than of c+/+
mice.15 Despite these data, it was possible that in the
special circumstances following the injection of endotoxin, this
situation might change and that an unusually high loss of GM-CSF in the urine might be responsible for the low serum levels of GM-CSF in
c+/+ mice.
To explore this possibility, urine was collected 3, 6, and 24 hours
after the intravenous injection of endotoxin. As shown in
Table 1, GM-CSF was detectible in the urine
of c+/+ mice at all timepoints analyzed in
concentrations that were higher than in the serum. With
c/ mice, GM-CSF was also detected in the
urine at all timepoints but in concentrations that were much higher
than in the urine of c+/+ mice and again in higher
concentrations than in the serum of the c/
mice.
Urinary loss of GM-CSF did not therefore provide an explanation of the
higher serum GM-CSF concentrations in c/
versus c+/+ mice after the injection of endotoxin,
because urinary loss of GM-CSF was in fact much higher in the
c/ mice than in the c+/+ mice.
G-CSF was not detectible in normal mouse urine but was readily
detectible following the injection of endotoxin. However in contrast to
the GM-CSF data, levels of G-CSF were similar in both types of mice at
the timepoints examined.
GM-CSF and G-CSF concentrations in peritoneal exudate fluid.
The injection interperitoneally to mice of casein containing nonviable
or saphrophytic bacteria induces a marked accumulation of neutrophils
within 3 hours and a rise in peritoneal fluid concentrations of GM-CSF
and G-CSF.18 In these earlier studies quantitative cellular
responses in the peritoneal cavity to the injection of casein were
found to be similar in c+/+ and
c/ mice.
As shown in Fig 5, a typical local CSF
response was observed in c+/+ mice following the
injection of casein. As noted in previous studies on normal
mice,18 GM-CSF responses occurred earlier than G-CSF
responses but declined after reaching peak values 2 hours after
injection.
View larger version (16K):
[in this window]
[in a new window]
| Fig 5.
GM-CSF and G-CSF concentrations in the peritoneal cavity
fluid of c+/+ and c/ mice at
intervals after the intraperitoneal injection of casein. Figures are
expressed as total nanograms of CSF in the 3-mL fluid volume collected
after harvesting. Each point is the mean value ± SD from two female
and one male mice of each genotype per timepoint.
|
|
c/ mice showed similar G-CSF responses to
the injection of casein but, in contrast to c+/+ mice,
GM-CSF responses were significantly higher and rose progressively during the 3-hour observation period, reaching a mean level of 20 ng at
3 hours, a level fivefold higher than in casein-injected c+/+ mice.
In Fig 5, the values are expressed as total nanograms CSF per
peritoneal cavity. In normal mice, the resident fluid volume has been
estimated as 70 µL,23 and in a real-life peritoneal infection with the accumulation of smaller fluid volumes than used in
the present model, the observed CSF levels would represent very high
local concentrations of CSF per milliliter fluid.
Production rates of GM-CSF by
c/ tissues.
The higher GM-CSF responses in c/ mice
than in c+/+ mice to the injection of endotoxin or
casein might have been based on a constitutively elevated capacity of
c/ tissues to produce GM-CSF.
This possibility was explored by examining the capacity of peritoneal
cells to produce CSF in short-term in vitro cultures after exposure to
casein and by determining the capacity of various organs to produce
GM-CSF and G-CSF in longer tissue cultures.
In the first approach, peritoneal cells from
c/ and c+/+ mice were
incubated for 3 hours with or without added casein, the concentrations of cells being 1 × 106 cells/mL to reduce the risk of
receptor-mediated removal of CSF. As shown in
Table 2, GM-CSF concentrations after 3 hours of incubation were elevated by addition of casein but the
concentrations were not significantly higher in cultures of
c/ cells than in cultures of
c+/+ cells. In cultures prepared containing 5 × 106 cells/mL, only a minor twofold higher concentration of
GM-CSF was observed in c/ versus
c+/+ cultures (data not shown), the minor difference
possibly reflecting the occurrence of some receptor-based clearance in
the c+/+ cultures.
An analysis of the capacity of various organs from
c/ and c+/+ mice to produce
GM-CSF after 4 days of incubation in vitro is shown in
Table 3. In agreement with previous
studies,19 individual organs varied widely in their
capacity to produce GM-CSF, but levels produced by
c/ organs did not significantly differ
from those produced by c+/+ organs.
Both sets of data suggested that c/ and
c+/+ tissues had an equivalent capacity to produce
GM-CSF under the conditions tested.
| |
DISCUSSION |
The present results document the fallacy of attempting to use serum CSF
concentrations to reach conclusions regarding the involvement or
noninvolvement of a CSF in responses to infections. The intravenous
injection of endotoxin was used as a surrogate model of a microbial
infection and was able to elicit a dramatic rise in serum G-CSF
concentrations in normal mice, but in contrast, only a minor response
in serum GM-CSF concentrations. These data paralleled earlier
observations on the lack of elevation of GM-CSF levels in
infections,10-13 observations that have led some to the conclusion that GM-CSF is likely to be of only minor importance in
systemic responses to infections.
The simple maneuver of using c/ mice,
whose cells lack high-affinity receptors for GM-CSF and have little
capacity to internalize GM-CSF, resulted in a very different outcome.
Following the injection of endotoxin, serum GM-CSF levels rose
approximately 30-fold above preinjection levels and achieved a mean
concentration of 9 ng/mL, a concentration in excess of that required to
stimulate maximal numbers of granulocyte-macrophage colonies to develop
in vitro. It is of interest that these GM-CSF responses always occurred slightly more rapidly than did G-CSF rises, with near-maximal responses
already being evident 1 hour after injection. The very rapid rise in
serum GM-CSF levels suggests that release of preformed GM-CSF to the
circulation may have been the basis for the initial rapid rise. An
increase in GM-CSF mRNA has been noted in macrophages after exposure to
endotoxin or other inducing agents in vitro, commencing within 1 to 2 hours.24 If this response pattern is typical for other cell
types, an increased synthesis of GM-CSF in vivo in response to
endotoxin, presumably therefore supervenes to maintain the observed
elevations in serum GM-CSF levels, because the half-life of GM-CSF in
serum is only 21minutes.15
The present experiments indicated that c/
and c+/+ tissues have a similar capacity to produce
GM-CSF and, while not investigating in detail the metabolic fate of
GM-CSF, eliminated abnormally rapid loss of GM-CSF in the urine of
endotoxin-injected c+/+ mice as a possible basis for
their failure to develop high serum GM-CSF levels.
The most reasonable conclusion is therefore that mice do in fact
respond promptly to challenge by endotoxin by elevating GM-CSF levels
but that this GM-CSF is usually efficiently removed by high-affinity
receptors, resulting in the apparent occurrence of only a very weak
response to challenge.
Earlier studies noted the production of GM-CSF locally in the
peritoneal cavity in response to the combined injection of casein and
nonviable or saphrophytic organisms as a model of local infection and
inflammation.18 The present studies showed that the
magnitude of this response is also underestimated in mice with
high-affinity GM-CSF receptors, not because of lower production rates
of GM-CSF but again presumably because of receptor-mediated endocytosis.
Because it is reasonable to suppose that hematopoietic cells are a
major receptor-bearing population in the body, the data further suggest
that these cells are likely to be very effectively stimulated by the
GM-CSF released or produced in response to endotoxin or local
infections. It therefore appears to be a mistake to regard GM-CSF as an
unresponsive regulator in the context of infections and indeed the
converse may be true, that it is an efficient and effective agent
during such responses. This conclusion is in line with the observations
that GM-CSF-deficient mice are prone to lung infections supervening on
their state of alveolar proteinosis7,8 and die prematurely
with a variety of infections.9
The present data reinforce previous observations19,21 that
dramatic serum G-CSF responses occur following the injection of
endotoxin, with levels reaching more than 1 µg/mL, a more than 1,000-fold rise above baseline levels. In view of the much lower concentrations of G-CSF that are adequate to elicit obvious
hematopoietic responses in vivo and in vitro, such extreme elevations
are remarkable and, if this type of response has a design purpose, it
will be of interest to determine what type of cellular responses
actually require such extreme concentrations.
The present data show that both GM-CSF and G-CSF levels respond
dramatically to the injection of endotoxin or local infections, but
with a differing pattern, local GM-CSF responses being higher and
systemic responses lower than G-CSF responses. The data from c/ mice suggest that much of the CSF
released or produced promptly binds to receptor-bearing cells. This
will lead to activation of the receptor-bearing cells and is thus a
rapid and effective signaling response able to activate appropriate
hematopoietic cells in response to microorganisms.
The phenomenon of receptor-mediated removal from the circulation of
GM-CSF is likely to apply equally to other comparable regulators and
should caution against overinterpretation of the significance of
observed regulator levels in the serum both in normal and abnormal situations.
| |
FOOTNOTES |
Submitted May 11, 1998; accepted October 29, 1998.
Supported by the Carden Fellowship Fund of the Anti-Cancer Council of
Victoria; the National Health and Medical Research Council, Canberra;
the AMRAD Corporation, Melbourne, Australia; and Grant No. CA-22556
from the National Institutes of Health, Bethesda, MD.
The publication costs of this
article were defrayed in part by
page charge payment. This article
must therefore be hereby marked
"advertisement"
in accordance with 18 U.S.C. section
1734 solely to indicate this fact.
Address reprint requests to Donald Metcalf, MD, The Walter and Eliza
Hall Institute of Medical Research, PO Royal Melbourne Hospital,
Victoria 3050, Australia.
| |
REFERENCES |
1.
Metcalf D:
The Hemopoietic Colony Stimulating Factors. Amsterdam, The Netherlands, Elsevier, 1984.
2.
Metcalf D, Nicola NA:
The Hemopoietic Colony Stimulating Factors. Cambridge, UK, Cambridge University Press, 1995.
3.
Sheridan JW, Stanley ER:
Tissue sources of bone marrow colony stimulating factor.
J Cell Physiol
78:451, 1971[Medline]
[Order article via Infotrieve]
4.
Metcalf D:
Acute antigen-induced elevation of serum colony stimulating factor (CSF) levels.
Immunology
21:427, 1971[Medline]
[Order article via Infotrieve]
5.
Sheridan JW, Metcalf D:
Studies on the bone marrow colony stimulating factor (CSF): Relation of tissue CSF to serum CSF.
J Cell Physiol
80:129, 1972[Medline]
[Order article via Infotrieve]
6.
Lieschke GJ, Grail D, Hodgson G, Metcalf D, Stanley E, Cheers C, Fowler KJ, Basu S, Zhan YF, Dunn AR:
Mice lacking granulocyte colony-stimulating factor have chronic neutropenia, granulocyte and macrophage progenitor cell deficiency, and impaired neutrophil mobilization.
Blood
84:1737, 1994[Abstract/Free Full Text]
7.
Stanley E, Lieschke GJ, Grail D, Metcalf D, Hodgson G, Gall JAM, Maher DW, Cebon J, Sinickas V, Dunn AR:
Granulocyte/macrophage colony-stimulating factor-deficient mice show no major perturbation of hematopoiesis but develop a characteristic pulmonary pathology.
Proc Natl Acad Sci USA
91:5592, 1994[Abstract/Free Full Text]
8.
Robb L, Drinkwater C, Metcalf D, Li R, Köntgen, Nicola NA, Begley CG:
Hematopoietic and lung abnormalities in mice with a null mutation of the common subunit of the receptors for granulocyte-macrophage colony-stimulating factor and interleukins 3 and 5.
Proc Natl Acad Sci USA
92:9565, 1995[Abstract/Free Full Text]
9.
Seymour JF, Lieschke GL, Grail D, Quilici C, Hodgson G, Dunn AR:
Mice lacking both granulocyte colony-stimulating factor (CSF) and granulocyte-macrophage CSF have impaired reproductive capacity, perturbed neonatal granulopoiesis, lung disease, amyloidosis, and reduced long-term survival.
Blood
90:3037, 1997[Abstract/Free Full Text]
10.
Cebon J, Layton JE, Maher D, Morstyn G:
Endogenous haemopoietic growth factors in neutropenia and infection.
Br J Haematol
86:265, 1994[Medline]
[Order article via Infotrieve]
11.
Watari S, Asano S, Shirafuji N, Kodo H, Ozawa K, Takaku F, Kamachi S:
Serum granulocyte colony-stimulating factor levels in healthy volunteers and patients with various disorders as estimated by enzyme immunoassay.
Blood
73:117, 1989[Abstract/Free Full Text]
12.
Kawakami M, Tsutsumi H, Kumakawa T, Abe H, Hirai M, Kurosawa S, Mori M, Fukushima M:
Levels of serum granulocyte colony-stimulating factor in patients with infections.
Blood
76:1962, 1990[Abstract/Free Full Text]
13.
Cheers C, Haigh AM, Kelso A, Metcalf D, Stanley ER, Young AM:
Production of colony-stimulating factors (CSFs) during infection: Separate determinations of macrophage-, granulocyte-, granulocyte-macrophage and Multi-CSFs.
Infect Immun
56:247, 1988[Abstract/Free Full Text]
14.
Kuter DJ, Rosenberg RD:
The reciprocal relationship of thrombopoietin (c-mpl ligand) to changes in the platelet mass during busulphan-induced thrombocytopenia in rabbits.
Blood
85:2720, 1995[Abstract/Free Full Text]
15.
Metcalf D, Mifsud S, Di Rago L, Robb L, Nicola NA, Alexander W:
The biological consequences of excess GM-CSF levels in transgenic mice also lacking high-affinity receptors for GM-CSF.
Leukemia
12:353, 1998[Medline]
[Order article via Infotrieve]
16.
Nicola NA, Peterson L, Hilton DJ, Metcalf D:
Cellular processing of murine colony-stimulating factor (Multi-CSF, GM-CSF, G-CSF) receptors by normal hemopoietic cells and cell lines.
Growth Factors
1:41, 1988[Medline]
[Order article via Infotrieve]
17.
Nicola NA, Metcalf D:
Binding internalization and degradation of 125I-multipotential colony-stimulating factor (interleukin-3) by FDCP-1 cells.
Growth Factors
1:29, 1988[Medline]
[Order article via Infotrieve]
18.
Metcalf D, Robb L, Dunn AR, Mifsud S, Di Rago L:
Role of granulocyte-macrophage colony-stimulating factor and granulocyte colony-stimulating factor in the development of an acute neutrophil inflammatory response in mice.
Blood
88:3755, 1996[Abstract/Free Full Text]
19.
Metcalf D, Willson TA, Hilton D, Di Rago L, Mifsud S:
Production of hematopoietic regulatory factors in cultures of adult and fetal mouse organs: Measurement by specific bioassays.
Leukemia
9:1556, 1995[Medline]
[Order article via Infotrieve]
20.
Metcalf D, Willson T, Rossner M, Lock P:
Receptor insertion into factor-dependent murine cell lines to develop specific bioassays for murine G-CSF and M-CSF and human GM-CSF.
Growth Factors
11:145, 1994[Medline]
[Order article via Infotrieve]
21.
Metcalf D, Roberts AW, Willson TA:
The regulation of hematopoiesis in max41 transgenic mice with sustained excess granulopoiesis.
Leukemia
10:311, 1996[Medline]
[Order article via Infotrieve]
22.
Metcalf D:
Mechanisms contributing to the sex difference in levels of granulocyte-macrophage colony-stimulating factor in the urine of GM-CSF transgenic mice.
Exp Hematol
16:794, 1988[Medline]
[Order article via Infotrieve]
23.
Gearing AJ, Metcalf D, Moore JG, Nicola NA:
Elevated levels of GM-CSF and IL-1 in the serum, peritoneal and pleural cavities of GM-CSF transgenic mice.
Immunol
67:216, 1989[Medline]
[Order article via Infotrieve]
24.
Thorens B, Mermod J-J, Vassalli P:
Phagocytosis and inflammatory stimuli induce GM-CSF mRNA in macrophages through post-transcriptional regulation.
Cell
48:671, 1987[Medline]
[Order article via Infotrieve]
 CiteULike  Connotea  Del.icio.us  Digg  Reddit  Technorati What's this?
This article has been cited by other articles:
|
|
|
|
 
S Saha, C Doe, V Mistry, S Siddiqui, D Parker, M Sleeman, E S Cohen, and C E Brightling
Granulocyte-macrophage colony-stimulating factor expression in induced sputum and bronchial mucosa in asthma and COPD
Thorax,
August 1, 2009;
64(8):
671 - 676.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
K. Uchida, K. Nakata, T. Suzuki, M. Luisetti, M. Watanabe, D. E. Koch, C. A. Stevens, D. C. Beck, L. A. Denson, B. C. Carey, et al.
Granulocyte/macrophage-colony-stimulating factor autoantibodies and myeloid cell immune functions in healthy subjects
Blood,
March 12, 2009;
113(11):
2547 - 2556.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
B. Larrivee, I. Pollet, and A. Karsan
Activation of Vascular Endothelial Growth Factor Receptor-2 in Bone Marrow Leads to Accumulation of Myeloid Cells: Role of Granulocyte-Macrophage Colony-Stimulating Factor
J. Immunol.,
September 1, 2005;
175(5):
3015 - 3024.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. Bozinovski, J. Jones, S.-J. Beavitt, A. D. Cook, J. A. Hamilton, and G. P. Anderson
Innate immune responses to LPS in mouse lung are suppressed and reversed by neutralization of GM-CSF via repression of TLR-4
Am J Physiol Lung Cell Mol Physiol,
April 1, 2004;
286(4):
L877 - L885.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. Bozinovski, J. E. Jones, R. Vlahos, J. A. Hamilton, and G. P. Anderson
Granulocyte/Macrophage-Colony-stimulating Factor (GM-CSF) Regulates Lung Innate Immunity to Lipopolysaccharide through Akt/Erk Activation of NFkappa B and AP-1 in Vivo
J. Biol. Chem.,
November 1, 2002;
277(45):
42808 - 42814.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
Top
Abstract
Introduction