|
|
 Previous Article | Table of Contents | Next Article 
Blood, Vol. 91 No. 9 (May 1), 1998:
pp. 3439-3446
Reduced Ex Vivo Interleukin-8 Production by Neutrophils in Septic
and Nonseptic Systemic Inflammatory Response Syndrome
By
Christelle Marie,
Jane Muret,
Catherine Fitting,
Marie-Reine Losser,
Didier Payen, and
Jean-Marc Cavaillon
From Unité d'Immuno-Allergie, Institut Pasteur, Paris; and the
Department of Anaesthesia and Intensive Care Medicine, Hôpital
Lariboisière, Paris, France.
 |
ABSTRACT |
Ex vivo cytokine production by circulating lymphocytes and monocytes
is reduced in patients with infectious or noninfectious systemic
inflammatory response syndrome. Very few studies have addressed the
reactivity of polymorphonuclear cells (PMN). To analyze further the
relative contribution of systemic inflammatory response syndrome alone
or in combination with infection we studied the interleukin-8 (IL-8)
production by PMN isolated from patients who had undergone cardiac
surgery with cardiopulmonary bypass (CPB) and patients with sepsis.
Cells were activated with either lipopolysaccharide (LPS) or
heat-killed streptococci. Compared with healthy controls, the release
of IL-8 by PMN in both groups of patients was significantly reduced
whether activated by LPS, independently of its concentration and
origin, or by heat-killed streptococci. These observations suggest that
stressful conditions related to inflammation, independently of
infection, rapidly dampened the reactivity of circulating PMN. We
investigated whether the observed diminished reactivity of PMN might
reflect an endotoxin tolerance phenomenon. Our in vitro experiments
with PMN from healthy controls indicated that PMN could not be rendered
tolerant stricto sensu. However, our data suggested that LPS-induced
mediators such as IL-10 may be responsible for the observed anergy in
patients.
| |
INTRODUCTION |
POLYMORPHONUCLEAR neutrophils (PMN) are
key cells in inflammatory processes and during sepsis syndrome. Their
activation is associated with the release of many inflammatory
mediators such as eicosanoids, free radicals, and proteolytic enzymes.
Activated neutrophils display an upregulated expression of CD11b and
CD351 and bind to endothelial cells, contributing to
vascular endothelium damage.2,3 They migrate toward tissues
where they maintain inflammation and favor organ
dysfunction,4-6 which may lead to lethality.7
The reactivity of neutrophils is modulated by many cytokines.
Pro-inflammatory cytokines such as interleukin-1 (IL-1), IL-8, and
tumor necrosis factor (TNF) are potent activators of PMN functions. The
fact that IL-8 is one of the main cytokines produced by
PMN8-10 allows these cells to perpetuate their own
activation within an autocrine loop. Anti-inflammatory cytokines play a
major role in dampening PMN functions, including IL-8 production. We
and others have shown that IL-10 and, to a lesser degree, IL-4, IL-13, and transforming growth factor- (TGF-), can repress IL-8 release by activated PMN.11-14 However, the nature of the
triggering signal influences the capacity of anti-inflammatory
cytokines to downmodulate cytokine release.14
During the course of inflammation, both pro- and anti-inflammatory
cytokines, rapidly released with excessive production, can be detected
in the blood compartment15 where they modulate the
reactivity of circulating leukocytes. Indeed, we have previously shown
that monocytes isolated from septic patients have a reduced capacity to
produce IL-1 , IL-1, IL-6, and TNF- upon activation by
endotoxins (lipopolysaccharide [LPS])16 or heat-killed
Gram-positive bacteria.17 This observation has been
confirmed by others with whole blood assays of the above
cytokines18,19 and extended to IL-10,20 IL-12,
and interferon- (IFN-),21 but not to IL-1 receptor
antagonist (IL-1ra) production.22 These observations probably follow the events occurring in the blood compartment, and
experiments with whole blood take into account the plasma environment,
which has been shown to play a major role in deactivating circulating
leukocytes.23 However, the source of cytokines cannot be
analyzed in whole blood assays. So far, McCall et al24 have been the only group to our knowledge to investigate the effect of
sepsis on PMN behavior in terms of cytokine production. They reported a
decreased capacity of neutrophils to produce IL-1 upon stimulation
in ex vivo experiments. The investigators suggested that the
observation was linked to the well-known endotoxin tolerance phenomenon. However, endotoxin tolerance is not initiated only by LPS
nor is it restricted to the LPS-responsiveness.25
Because IL-8 is the most abundant cytokine produced by neutrophils, we
were interested in analyzing the capacity of PMN from septic patients
to produce this specific chemokine. It was of interest to compare the
observable modifications in septic patients consecutive to the
infectious process itself and those associated with the inflammatory
reaction, independently of any microbial stimulus. Indeed, we had
previously shown that inflammation after surgery was sufficient to
reduce the capacity of monocytes to produce TNF-, IL-1, and
IL-1.26 For that purpose we investigated a group of
patients who had undergone cardiac surgery associated with
cardiopulmonary bypass (CPB). It is well known that, although surgery
itself is an inflammatory process, the extracellular circulation is
associated with an enhanced inflammatory reaction as a consequence of
the interaction of blood with bio-materials.27 Finally, we attempted to mimic some of the events occurring in vivo to find out
whether endotoxin and/or IL-10 could be responsible for the modifications we observed in both groups of patients.
| |
MATERIALS AND METHODS |
Patients.
Eleven patients with sepsis syndrome, as defined by Bone et
al,28 were studied upon admission to intensive care units
(ICU) or at initiation of their sepsis syndrome. The study included 9 men and 2 women, the average age being 49 ± 5 years (range, 21 to 69 years) and the mean simplified acute physiology score II (SAPS II)
being 49 ± 7. There were 8 cases of pneumonia, 1 of pelvic abscess,
and 2 of peritonitis, including 5 in septic shock. The outcome was
survival for 4 patients. The patients' characteristics are given in
Table 1.
Nine patients undergoing cardiopulmonary bypass for selective artery
bypass grafting were studied. The study included 8 men and 1 woman, the
average age being 68 ± 2 years (range, 63 to 74 years). Anesthesia
induction and maintenance was achieved using flunitrazepam, sufentanyl,
pancuronium bromide, and isoflurane. Administration of heparin before
canulation and subsequent neutralization after bypass with protamine
sulfate were performed in a standard fashion. Normothermic CPB was
performed (32.5 ± 0.4°C). The mean timing of aortic clamping was
55 ± 2 minutes. Blood sampling was performed 11/2 hours
after CPB beginning and always before protamine administration.
Blood of healthy controls were obtained from 18 subjects either from
the blood bank (Fondation Nationale de Transfusion Sanguine) or from
the laboratory. The study included 7 men and 11 women, the average age
being 32 ± 2 years (range, 24 to 51 years).
Isolation of human polymorphonuclear cells.
Blood was drawn onto heparin (20 IU/mL) from healthy volunteers or from
patients. Ten volumes of blood was mixed with 2 vol of glucose dextran
(3% glucose; 3% dextran T250; Pharmacia, Uppsala, Sweden) and the
leukocytes were recovered after a 40-minute sedimentation at room
temperature.29 The leukocytes were then diluted 1:2 in
RPMI-1640 medium and layered on Ficoll-Hypaque (Milieu de
Séparation des Lymphocytes [MSL]; Eurobio, Les Ulis, France).
The ratio was 2 vol of leukocytes to 1 vol of MSL. After centrifugation
for 25 minutes at 15°C and 500g, the cell pellet was washed
and centrifuged once for 5 minutes at 300g. Contaminating
erythrocytes were lysed after a 5-minute incubation of the cell pellet
at 4°C by resuspension in 5 mL of lysis buffer
(NH4Cl = 8.32 g/L; NaHCO3 = 0.84 g/L; Na4EDTA = 43.2 mg/L). Lysis was stopped by adding a large
excess of RPMI-1640 medium (Glutamax; GIBCO Life Technologies, Paisley, UK) and the cells were washed and centrifuged for 10 minutes at 200g. The viability of polymorphonuclear cells (PMN)
was assessed by counting the cells in 0.1% eosine. A nonspecific
esterase staining was performed to evaluate the monocyte contamination,
which never exceeded 0.5%.
In vitro culture.
PMN cells were cultured in RPMI-1640 medium (Glutamax; GIBCO Life
Technologies) supplemented with antibiotics (penicillin, 100 IU/mL;
streptomycin, 100 µg/mL) and 5% heat-inactivated normal human serum
(a pool of sera from healthy volunteers). 0.5-mL aliquots of PMN
suspension per well were incubated in a 5% CO2 incubator in 24-well multidish plates (Costar, Cambridge, MA) for 24 hours at
37°C. Stimuli in volumes  10 µL were added at the beginning of the
culture. At the end of the culture, the supernatants were obtained,
centrifuged for 10 minutes at 300g and 15°C, and kept at
 20°C before cytokine assessments.
Reagents.
Escherichia coli (0111:B4) lipopolysaccharide was purchased
from Sigma (St Louis, MO), TNF- was obtained from Rhône
Poulenc (Vitry/Seine, France), and Neisseria meningitidis LPS
was the generous gift of Dr Martine Caroff (Institut de Biochimie,
Orsay, France). Heat-killed Streptococcus pyogenes group A
(A78) was a kind gift of Dr H. Müller-Alouf
(Institut Pasteur de Lille). Recombinant human IL-10 was a
generous gift of Dr J. Abrams (DNAX, Palo Alto, CA).
Two-step experiments.
PMN (2 × 106 cells/mL) were cultured in RPMI-1640
medium (Glutamax; GIBCO Life Technologies) supplemented with
antibiotics and 5% heat-inactivated normal human serum (complete
medium). A 0.5-mL aliquot of PMN suspension per well in 24-well
multidish plates was incubated for 24 hours at 37°C in a 5%
CO2 incubator in the presence of different reagents
("pretraitment period"). At the end of this preculture period,
the supernatants were obtained and 0.25 mL of fresh complete medium was
added to each well to preserve adherent neutrophils. Supernatants were
centrifuged for 10 minutes at 200g and 15°C and kept at
20°C before IL-8 assessment. The cell pellets were resuspended in
0.25 mL of fresh complete medium and plated in the same well as during
the first 24 hours. Then, multidish plates were incubated at 37°C for
an additional 24 hours of culture in the presence of various activators
("activation period"). At the end of the activation period, the
supernatants were obtained, centrifuged 10 minutes at 300g and
15°C, and kept at 20°C before IL-8 assessment.
Assessement of cell-associated forms of IL-8 in whole-blood samples.
One milliliter of blood was centrifuged for 10 minutes at 500g.
Plasma was obtained and kept at 20°C before cytokine assessments. The cell pellet was lysed in 100 µL of lysis buffer (TRAx buffer; T-Cell Sciences, Inc, Needham, MA) before addition of RPMI-1640 medium
to achieve a 1-mL total volume.
Assessement of cell-associated forms of IL-8 in PMN.
At the end of the culture period (2 × 106 PMN/mL), the
PMN pellets were lysed by adding 100 µL of TRAx lysis buffer before adding 100 µL of diluent buffer and 300 µL of RPMI medium.
Cytokine enzyme-linked immunosorbent assay (ELISA).
IL-8 ELISA was performed as previously described30 using a
monoclonal anti-human IL-8 antibody obtained by Dr J-C. Mazié (Hybridolab, Institut Pasteur) and a rabbit polyclonal anti-IL-8 antibody graciously provided by Dr N. Vita (Sanofi Recherche, Labège, France). The sensitivity of the ELISA was 3 pg/mL. In collaboration with Dr S. Berthold (DPC Biermann, Bad Nauheim, Germany)
the values obtained with our ELISA were compared with those obtained
for the same samples using the chemiluminescent quantification of IL-8
(IMMULITE; DPC Biermann). A correlation of r = .98 (P = .0001) was achieved. We ensured that the TRAx buffer did
not interfere with the accuracy of the ELISA.
Apoptosis assay.
PMN apoptosis was assessed as the percent of cells with hypodiploid DNA
by using the technique described by Nicoletti et al.31 After 24 or 48 hours of culture of PMN with or without activators, cells were centrifuged at 200g for 10 minutes and washed in
phosphate-buffered saline (PBS). The cell pellets were gently
resuspended in hypotonic fluorochrome solution (50 µg/mL propidium
iodide [PI], 0.1% sodium citrate, 0.1% Triton X-100) and stored at
4°C in the dark overnight before the flow-cytometric analysis using a
FACScan flow cytometer (Becton Dickinson Immunocytometry System, San
Jose, CA). The red fluorescence of PI in individual nuclei and the
forward and side scatter were simultaneously measured. Cell debris were
excluded from acquisition by raising the forward scatter threshold.
Apoptotic nuclei were easily distinguishable from residual debris by
the high size side scatter value due to the condensation of nuclear chromatin. Ten thousand events were collected and analyzed using the
software Lysis II program. Apoptotic PMN nuclei were distinguished by
their hypodiploid DNA content from the diploid DNA content of normal
PMN nuclei.
Statistical analysis.
Statistical analyses were performed using the nonparametric
Mann-Whitney U-test for comparing data between healthy controls and
patients and the Wilcoxon signed-rank test for the two-step experiments
performed with the PMN from healthy controls.
| |
RESULTS |
Circulating and cell-associated IL-8.
Levels of circulating IL-8 were under the detection limit in controls
and in all but two patients undergoing cardiac surgery before
cardiopulmonary bypass (Fig 1), whereas
after cardiopulmonary bypass all patients but two had detectable levels
of plasma IL-8. Similarly, IL-8 was measurable in all but two septic
patients. The highest values were found among septic patients but the
mean levels between post-CPB and sepsis were not significantly
different.
View larger version (18K):
[in this window]
[in a new window]
| Fig 1.
Measurement of circulating IL-8 in plasma (left) of
healthy controls, patients undergoing cardiac surgery before (pre-CPB), and after (post-CPB) cardiopulmonary bypass and patients with sepsis.
Cell-associated IL-8 was assessed in whole-blood cell pellet of the
same groups of donors (right). Each symbol represents an individual
subject and the thick short lines represent the median value for each
group.
|
|
The pattern was quite different for cell-associated IL-8. We have
previously shown that cell-associated IL-8 assessed in whole blood is
essentially associated with leukocytes whereas the contribution of red
blood cells is limited.32 Naturally occurring
cell-associated IL-8 was detected in healthy controls, and levels were
similar in pre-CPB patients. Post-CPB patients had significantly higher levels than those measured before CPB (P = .015). The levels
of cell-associated IL-8 in septic patients were also higher than in
controls as well as higher than in post-CPB patients
(P = .007). Although circulating levels of IL-8 were similar
in infectious and noninfectious inflammation, the levels of
cell-associated IL-8 were not.
IL-8 production by isolated PMN.
We studied the capacity of isolated circulating PMN to release IL-8 in
response to either 0.1 or 1 µg/mL of LPS of two different bacterial
origins or heat-killed streptococci (Fig
2). A significant reduction in the levels
of IL-8 produced in response to the different concentrations of LPS
from different origins and to heat-killed streptococci was observed in
both groups of patients. Sepsis or infectious systemic inflammatory
response syndrome as well as noninfectious inflammation both led to a
reduced capacity of PMN to release IL-8 upon in vitro stimulation. IL-8
production in patients undergoing cardiac surgery was also tested in a
few patients before the initiation of the cardiopulmonary bypass, and
the levels of released IL-8 were similar to those obtained in healthy
controls (data not shown).
View larger version (25K):
[in this window]
[in a new window]
| Fig 2.
IL-8 production by isolated polymorphonuclear cells (PMN)
after in vitro activation with LPS from different origins
(Escherichia coli: E.c.; Neisseria meningitidis: N.m.)
or with heat-killed, group A streptococcus. The data are the mean of
experiments performed with isolated PMN from healthy controls
(n = 13), post-CPB patients (n = 9), and patients with sepsis
(n = 11). P values correspond to statistical analysis
between post-CPB or sepsis groups versus control one.
|
|
After cell activation, the levels of IL-8 were also analyzed at the
cellular levels, ie, following cell lysis of PMN (Table 2).
There was no difference between the levels of
cell-associated IL-8 in septic and control groups whereas in most cases
there was a significant reduction of the levels found in post-CPB
patients.
In vitro modulation of IL-8 production by PMN from healthy donors.
The reduced levels of IL-8 release by PMN isolated from septic patients
or patients who underwent cardiopulmonary bypass could be the
consequence of in vivo exposure of PMN to endotoxin and/or to
IL-10. We attempted to mimic this situation by preculturing PMN from
healthy donors in the presence of LPS, IL-10, or both, before further
activation by LPS. Numerous investigators have reported a high percent
of apoptotic PMN after a 24-hour culture period.33,34
However, our PMN preparations (dextran + glucose sedimentation and a
Ficoll-Hypaque [MSL] step) and our culture conditions in the presence
of normal human serum led to a low percentage of apoptotic cells after
a 24-hour culture period. Recovery of live cells after an overnight
culture period was 89% ± 14% with 15% ± 2% and 9% ± 2%
apoptotic cells in the absence or in the presence of LPS, respectively.
After 48 hours the recovery was 46% ± 15% live cells and 35% ± 11% apoptotic cells. Thus, it was possible to perform the
two-step experiments over 48 hours. The mean values of four experiments
are shown in Fig 3: PMN maintained for 24 hours in culture medium alone were fully responsive to LPS during an
additional 24-hour culture period and release significant amounts
of IL-8 in a dose-dependent fashion, illustrating the good viability of
the cells. Such IL-8 production (24 to 48 hours) also occurred when the
cells had been precultured in the presence of increasing amounts of LPS
and then left in cultured medium alone. LPS-pretreated cells responded
in an additional fashion when reexposed to LPS. Similarly,
LPS-pretreated PMN had an enhanced IL-8 production upon a further
challenge by recombinant human TNF (10 ng/mL) compared with cells
maintained for 24 hours in culture medium alone (8,631 ± 3,880
v 3,293 ± 925, P = .04; mean of five
experiments). Thus, in contrast to monocytes which upon preexposure to
LPS are "tolerized" and have a lower capacity to release
cytokines,35 PMN were not rendered tolerant to LPS by prior
encounter with LPS. To see whether this effect was specific for LPS or
could be obtained with other activating signals, PMN were first
cultured in the presence of TNF, and further cultured in the presence
of LPS (Table 3). TNF pretreated PMN
continue to release IL-8 once TNF has been withdrawn. Addition of LPS
acted synergistically and led to an enhanced release of IL-8, while a
second TNF stimulation had an insignificant enhancing effect.
View larger version (86K):
[in this window]
[in a new window]
| Fig 3.
In vitro model of tolerization. PMN from healthy donors
were first cultured over 24 hours in the presence or absence of
increasing amounts of E coli LPS. Supernatants were obtained
and fresh medium was added to the cells. PMN were further cultured for
an additional 24 hours (24-48 h) in the presence or absence of
increasing amounts of E coli. IL-8 was measured in the
supernatants. The results are the mean of four different experiments;
the mean of SEM was ±30%.
|
|
To further understand the mechanism leading to the PMN anergy we
observed in the patients, we analyzed the effects of IL-10, known to
circulate in these patients. As shown in Fig 4A,
simultaneous addition of IL-10 and LPS led to an 88%
inhibition of IL-8 production by PMN, confirming numerous previous
studies, including ours.11,14 Pretreatment of PMN with
IL-10 render the cells less reactive to a subsequent stimulation with
LPS (P = .02) (Fig 4B). Furthermore, pretreatment of PMN with
IL-10 abolished the priming effect of LPS to a further activation by
LPS (P = .01).
View larger version (24K):
[in this window]
[in a new window]
| Fig 4.
In vitro model of tolerization. PMN from healthy donors
were first cultured over 24 hours in the presence or absence of either E coli LPS (100 ng/mL) or recombinant human IL-10 (10 ng/mL) or both. Supernatants were obtained and IL-8 was measured (A). Fresh medium was added to the cells and the PMN were further cultured for an
additional 24-hour period (24-48 h) in the presence or absence of E
coli LPS (1 µg/mL). IL-8 was measured in the supernatants (B).
The results are the mean of eight different experiments. (b v
a: P = .01; c v a: P = .02; d v
a: P = .01; d v b: P = .01.)
|
|
| |
DISCUSSION |
Circulating and cell-associated IL-8.
Systemic inflammatory response syndrome of infectious (eg, sepsis
syndrome) or noninfectious (eg, cardiopulmonary bypass surgery) origin
can both be associated with the detection of circulating cytokines.
Although the plasma levels of IL-8 could not discriminate between the
two types of inflammation, significantly higher levels of
cell-associated IL-8 were measured in septic patients (Fig 1). An
enhanced cell-associated IL-8 was found in patients after cardiac
surgery involving CPB as compared to pre-CPB in agreement with a
previous report.36 We have previously shown that a high proportion of IL-8 in the blood compartment was associated with erythrocytes and mainly with mononuclear and polymorphonuclear cells.32 A major proportion of the cell-associated IL-8
found with leukocytes was the result of internalization of surrounding IL-8. Because post-CPB and septic patients had similar levels of
circulating PMN (data not shown), the relative number of circulating cells may have had little influence on the lower level of
cell-associated IL-8 observed in the CPB group. These results may
rather reflect lower IL-8 receptor expression and IL-8 internalization
process in this group of patients. The analysis of cell-associated IL-8 after in vitro activation which led to a diminished level of
PMN-associated IL-8 among CPB patients as compared with healthy
controls (Table 2) further argues for a lower IL-8 receptor expression.
Indeed, C5a generated during extracorporal circulation and IL-8 itself known to modulate IL-8 receptor expression37,38 may be the causative agents which led to low IL-8 receptor expression. Further experiments will test it.
Among the septic patients, the reduced levels of IL-8 in PMN
supernatants was not associated with reduced levels of the
cell-associated form as compared with healthy donors. We can speculate
that the receptor-mediated trapping of the IL-8 released by PMN upon
activation and its internalization was achieved with a certain amount
of IL-8, a level of which was reached in both healthy and
septic patients. The PMN of healthy donors do produce more IL-8 than those from septic patients and the difference could only be found in
the PMN supernatants.
Sepsis and reduced cytokine production/PMN hyporeactivity.
Whole-blood assays have been widely used these last years to
investigate the capacity of circulating leukocytes to produce cytokines
within their local environment. A reduced capacity was observed when
IL-1, TNF-, IL-6, IL-10, IL-12, and IFN- were investigated.18-22,39,40 We also studied the capacity of
whole-blood samples to generate IL-8 upon in vitro activation. Low
concentrations of endotoxins, independently of their origin, allow the
detection of a diminished capacity to produce IL-8 in septic patients
compared with healthy controls. On the contrary, when higher amounts of LPS or heat-killed streptococci were used, no significant changes were
observed. In CPB patients the capacity to produce IL-8 was unchanged as
compared to controls (data not shown).
Experiments performed with whole-blood assays give no information about
the specific reactivity of a given cell lineage. We have previously
investigated the capacity of monocytes from septic patients to be
activated.16,17 Because IL-8 is mainly produced by both
monocytes and neutrophils within the blood compartment we decided to
analyze the PMN reactivity. Indeed, very few studies have investigated
the production of cytokines by neutrophils from septic patients and we
showed that, as previously reported for IL-1,24
circulating PMN have a significantly reduced capacity to release IL-8.
In contrast to the data reported on IL-1, our observation was made
independently of the nature of the triggering signal. Endotoxins from
various origins used at different concentrations as well as heat-killed
streptococci led to a reduced production of IL-8 compared with healthy
subjects.
Although PMN have been claimed to exist in one of three states
(quiescent, primed, or activated),41 it is clear that a
fourth state, ie, deactivated, exists. Indeed, other PMN functions have been reported to be reduced in infected patients such as the protein kinase C-dependent stimulation of superoxide production.42
In rabbits, in vitro complement (C5)-dependent degranulation of PMN was
reduced after intravenous injection of LPS,43 and in human volunteers administration of endotoxin leads to a markedly reduced ex
vivo neutrophil chemotactic activity.44 Accordingly, in in vivo experiments, LPS-pretreated animals showed a diminished
recruitment of PMN into tissues after local challenge.43,45
Deactivation of PMN has also been reported in noninfectious
pathologies. For example, Parsons et al46 reported that PMN
from patients with adult respiratory distress syndrome (ARDS) produced
less superoxide than did cells from normal subjects when primed with
LPS and stimulated with formyl-methionyl-leucine-phenylalanine.
However, not all PMN functions may be altered during inflammation
and/or infection because H2O2
generation was shown to be enhanced in septic patients47 and in a PMN subpopulation from ARDS patients.48
Inflammatory stress and ex vivo cytokine production.
To see whether the reduced IL-8 production by PMN we observed in septic
patients was specific to an infectious process or related to the
systemic inflammatory response, we included another group of patients
who underwent cardiac surgery associated with cardiopulmonary bypass.
Inflammatory stress that occurs during CPB is linked to the surgical
procedure itself as well as to the interaction of circulating
leukocytes with biomaterials during extracorporal
circulation.27 To our knowledge, very few investigators have tested the ex vivo capacity of leukocytes of CPB patients to
release cytokine upon activation. Naldini et al49 have
reported that phytohemagglutinin-induced production of IFN-, IL-2,
and TNF by peripheral blood mononuclear cells were significantly
diminished while IL-1, IL-6, and IL-8 were not affected. In
addition, we have previously shown that surgery itself leads to a
reduced capacity of monocytes to release IL-1, IL-1, and TNF-,
but not IL-6.26 Other stressful conditions such as
trauma,21,50,51 thermal injury,52 and
hemorrhage53 are also associated with a reduced capacity of
leukocytes to produce cytokine upon cell stimulation. We observed that
IL-8 production by PMN as well as IL-10 production by peripheral blood
mononuclear cells (manuscript submitted) were significantly reduced in
CPB patients. In addition to the inflammatory stress, other parameters
such as some drugs may affect cytokine production by circulating
leukocytes. Among the drugs used in the patients, aprotinin, a serine
protease inhibitor, has been shown to reduce IL-8 levels in
broncho-alveolar lavages of patients after CPB.54 We have
performed experiments by adding aprotinin to whole blood of healthy
controls at concentrations similar to those generated in the patients'
blood (500 KIU/mL). Blood was maintained at 37°C for 3 hours before
performing the PMN preparation, similarly to the in vivo situation
before sampling the blood from the post-CPB patients. After PMN
activation by either LPS or streptoccoci, an enhanced release of IL-8
was observed (49% ± 9%) (data not shown), just at the
opposite of the ex vivo observation. This observation
indicates that aprotinin is not the causative agent of the decreased
IL-8 production observed in CPB patients. This does not mean that other
drugs such as those used for anesthesia do not interfere in the ability
of PMN to produce IL-8. However, in a preliminary study on four
patients undergoing prolonged anesthesia with a light surgical stress
(arterial embolization for aneurysm), we did not observe any
significant modification of the IL-8 production by isolated PMN upon
activation.
The presence of circulating pro- and anti-inflammatory cytokines such
as IL-6,55 IL-8,55,56 TNF,57
IL-10,58 TGF-,59 as well as the induction of
cell-associated IL-160 in patients undergoing CPB, confirms
that a systemic inflammatory process does occur. Thus, a rapid
deactivation of circulating PMN in terms of IL-8 production was
observed in CPB patients, suggesting that the inflammatory component of
sepsis syndrome may be sufficient to alter the IL-8 synthesis machinery
in PMN. Although CPB is not associated with infectious processes,
circulating LPS has been detected in these patients61 and,
as in infected patients, its role in affecting PMN reactivity could be
considered as endotoxin tolerance.25
In vitro tolerization of PMN from healthy donors.
To determine whether an LPS encounter in vivo in patients could alter
PMN reactivity to a later in vitro LPS stimulation, we performed
two-step experiments with PMN prepared from healthy controls. PMN were
first stimulated with LPS overnight and resuspended in fresh medium in
the presence of activating LPS for another 24-hour culture. IL-8
production was then assessed in the cell supernatants. Although a high
percentage of PMN has been reported to become rapidly apoptotic in
cultures,33,34 our culture conditions had a low level of
apoptotic cell after 24 hours and allowed us to perform these two-step
analyses. Although a similar protocol led to tolerized
monocytes,35,62 neutrophils that were first exposed to LPS
before a secondary stimulation with LPS had an enhanced cytokine
production.
In monocytes, IL-10 could mimic in vitro LPS
tolerization,35 and septic63 as well as
CPB58 patients have circulating IL-10. So, it was
worthwhile to see whether pretreatment of PMN by IL-10 led to a
reduction of IL-8 production. As previously reported,11,14
IL-10 is a potent inhibitor of IL-8 production by PMN when added
simultaneously with LPS. The effect of pretreatment of PMN by IL-10 led
to a significant reduction of subsequent LPS-induced IL-8 production. Furthermore, when IL-10 and LPS were added simultaneously during the
pretreatment period, IL-10 completely obliterated the "priming" effects of LPS. Thus, while PMN can be rendered less reactive to a
secondary stimulation by LPS after pretreatment with IL-10, they cannot
be rendered tolerant to endotoxin stricto sensu by LPS itself. McCall
et al24 claimed that PMN from septic patients were
endotoxin tolerant. However, this statement should be reconsidered very
carefully. Although there are many earlier reports that PMN could be
primed by a short preexposure to LPS for enhanced release of oxygen
metabolites64 and for the synthesis of 5-lipoxygenase products65 there was no long exposure of PMN and, to our
knowledge, this is the first demonstration of a priming effect of LPS
on IL-8 production. A similar priming effect was noticed after a first
encounter of PMN with TNF. These results obtained after a first
activation of long duration are in contrast with a previous report66 in which a 2-hour pretreatment with low amounts of TNF led to deactivated PMN with reduced transcobalamine release and
burst activity in response to TNF. In this report, in contrast to most
other reports, a 2-hour pretreatment with LPS also led to a
deactivation of PMN when further challenged with TNF.
We have focused our attention on the responsiveness of isolated
neutrophils from patients with systemic inflammatory response syndrome
of infectious or noninfectious origin. In both cases the release of
IL-8 upon activation by endotoxins or heat-killed streptococci was
significantly reduced, suggesting that stressful conditions rapidly
dampen the reactivity of circulating PMN. As suggested by in vitro
experiments performed with PMN from healthy controls, the observed
phenomenon does not reflect an endotoxin tolerance phenomenon stricto
sensu because LPS was unable to desensitize the cells to a second
challenge by LPS as reported for monocytes. Our in vitro
results suggest that LPS-induced mediators such as IL-10 may be
responsible for the observed anergy in patients.
| |
FOOTNOTES |
Submitted October 1, 1997;
accepted December 22, 1997.
Address reprint requests to Jean-Marc Cavaillon, DrSc, Unité
d'Immuno-Allergie, Institut Pasteur, 28 rue Dr Roux, 75015 Paris, France.
The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" is accordance with 18 U.S.C. section 1734 solely to indicate this fact.
| |
ACKNOWLEDGMENT |
We thank the Cardiopulmonary Unit of the Hôpital
Lariboisière for its support and help, and Dr Dorris Grossmann
for linguistic advice.
| |
REFERENCES |
1.
Rosenbloom AJ,
Pinsky MR,
Bryant JL,
Shin A,
Tran T,
Whiteside T:
Leukocyte activation in the peripheral blood of patients with cirrhosis of the liver and SIRS. Correlation with serum interleukin-6 levels and organ dysfunction.
JAMA
274:58,
1995[Abstract/Free Full Text]
2.
Westlin WF,
Gimbrone MA Jr:
Neutrophil-mediated damage to human vascular endothelium. Role of cytokine activation.
Am J Pathol
142:117,
1993[Abstract]
3.
Fabian TC,
Crose MA,
Stewart RM,
Dockter ME,
Proctor KG:
Neutrophil CD18 expression and blockade after traumatic shock and endotoxin challenge.
Ann Surg
220:552,
1994[Medline]
[Order article via Infotrieve]
4.
Hewett JA,
Schultze AE,
VanCise S,
Roth RA:
Neutrophil depletion protects against liver injury from bacterial endotoxin.
Lab Invest
66:347,
1992[Medline]
[Order article via Infotrieve]
5.
Gossage JR,
Kuratomi Y,
Davidson JM,
Lefferts PL,
Snapper JR:
Neutrophil elastase inhibitors, SC-37698 and SC-39026 reduce endotoxin-induced lung dysfunction in awake sheep.
Am Rev Respir Dis
147:1371,
1993[Medline]
[Order article via Infotrieve]
6.
Mallick AA,
Ishizaka A,
Stephens KE,
Hatherill JR,
Tazeloer HD,
Raffin TA:
Multiple organ damage caused by tumor necrosis factor and prevented by prior neutrophil depletion.
Chest
95:1114,
1989[Abstract/Free Full Text]
7.
Nuijens J,
Abbink J,
Wachtfogel Y,
Colman R,
Eerenberg A,
Dors D,
Kamp A,
Strack Van Schijndel R,
Thijs L,
Hack C:
Plasma elastase 1-antitrypsin and lactoferrin in sepsis: Evidence for neutrophils as mediators in fatal sepsis.
J Lab Clin
119:159,
1992[Medline]
[Order article via Infotrieve]
8.
Strieter RM,
Kasahara K,
Allen RM,
Standiford TJ,
Rolfe MW,
Becker FS,
Chensue SW,
Kunkel SL:
Cytokine-induced neutrophil-derived interleukin-8.
Am J Pathol
141:397,
1992[Abstract]
9.
Cassatella MA,
Bazzoni F,
Ceska M,
Ferro I,
Baggiolini M,
Berton G:
IL-8 production by human polymorphonuclear leukocytes. The chemoattractant formyl-methionyl-leucyl-phenylalanine induces the gene expression and release of IL-8 through a pertussis toxin-sensitive pathway.
J Immunol
148:3216,
1992[Abstract]
10.
Altstaedt J,
Kirchner H,
Rink L:
Cytokine production of neutrophils is limited to interleukin-8.
Immunology
89:563,
1996[Medline]
[Order article via Infotrieve]
11.
Cassatella MA,
Meda L,
Bonora S,
Ceska M,
Constantin G:
Interleukin 10 (IL-10) inhibits the release of proinflammatory cytokines from human polymorphonuclear leukocytes. Evidence for an autocrine role of tumor necrosis factor and IL-1 in mediating the production of IL-8 triggered by lipopolysacharide.
J Exp Med
178:2207,
1993[Abstract/Free Full Text]
12.
Kasama T,
Strieter RM,
Lukacs NW,
Burdick MD,
Kunkel SL:
Regulation of neutrophil derived chemokine expression by IL-10.
J Immunol
152:3559,
1994[Abstract]
13.
Wertheim WA,
Kunkel SL,
Standiford TJ,
Burdick MD,
Becker FS,
Wilke A,
Gilbert AR,
Strieter RM:
Regulation of neutrophil-derived IL-8: The role of prostaglandin E2, dexamethasone and IL-4.
J Immunol
151:2166,
1993[Abstract]
14.
Marie C,
Pitton C,
Fitting C,
Cavaillon J-M:
Regulation by anti-inflammatory cytokines (IL-4, IL-10, IL-13, TGF) of interleukin-8 production by LPS- and/or TNF-activated human polymorphonuclear cells.
Mediators Inflam
5:334,
1996
15.
Cavaillon JM,
Muñoz C,
Fitting C,
Misset B,
Carlet J:
Circulating cytokines: The tip of the iceberg?
Circ Shock
38:145,
1992[Medline]
[Order article via Infotrieve]
16.
Muñoz C,
Carlet J,
Fitting C,
Misset B,
Bleriot JP,
Cavaillon JM:
Dysregulation of in vitro cytokine production by monocytes during sepsis.
J Clin Invest
88:1747,
1991
17. Cavaillon J-M, Muñoz C, Marty C, Cabié A, Tamion F,
Misset B, Carlet J, Fitting C: Cytokine production by monocytes from
patients with sepsis syndrome and by endotoxin-tolerant monocytes in
Levin J, Alving CR, Munford RS, Stütz PL (eds): Bacterial Endotoxin: Recognition and Effector Mechanisms. Amsterdam, The Netherlands, Elsevier, 1993, p 275
18.
Sekatrian JC,
Yee J,
Christou NV:
Reduced tumor necrosis factor- production in lipopolysaccharide-treated whole blood from patients in the intensive care unit.
Arch Surg
129:187,
1994[Abstract/Free Full Text]
19.
Döcke WD,
Randow F,
Syrbe U,
Krausch D,
Asadullah K,
Reinke P,
Volk HD,
Kox W:
Monocyte deactivation in septic patients: Restoration by IFN treatment.
Nature Med
3:678,
1997[Medline]
[Order article via Infotrieve]
20.
Marchant A,
Alegre M,
Hakim A,
Piérard G,
Marécaux G,
Friedman G,
De Groote D,
Kahn R,
Vincent J,
Goldman M:
Clinical and biological significance of interleukin-10 plasma levels in patients with septic shock.
J Clin Immunol
15:265,
1995
21.
Ertel W,
Keel M,
Neidhardt R,
Steckholzer U,
Kremer JP,
Ungethuem U,
Trentz O:
Inhibition of the defense system stimulating interleukin-12 interferon- pathway during critical illness.
Blood
89:1612,
1997[Abstract/Free Full Text]
22.
Van Deuren M,
Van Der Ven-Jongekrijg H,
Demacker PNM,
Baterlink AKN,
Van Dalen R,
Sauerwein RW,
Gallati H,
Vannice J,
van Der Meer JWM:
Differential expression of proinflammatory cytokines and their inhibitors during the course of meningococcal infections.
J Infect Dis
169:157,
1994[Medline]
[Order article via Infotrieve]
23.
Brandtzaeg P,
Osnes L,
Øvstebø R,
Joø GB,
Westwik AB,
Kierulf P:
Net inflammatory capacity of human septic shock plasma evaluated by a monocyte-based target cell assay: Identification of interleukin-10 as a major functional deactivator of human monocytes.
J Exp Med
184:51,
1996[Abstract/Free Full Text]
24.
McCall CE,
Grosso-Wilmoth LM,
LaRue K,
Guzman RN,
Cousart SL:
Tolerance to endotoxin-induced expression of the interleukin-1 gene in blood neutrophils of humans with the sepsis syndrome.
J Clin Invest
91:853,
1993
25.
Cavaillon JM:
The nonspecific nature of endotoxin tolerance.
Trends Microbiol
3:320,
1995[Medline]
[Order article via Infotrieve]
26.
Cabié A,
Fitting C,
Farkas J-C,
Laurian C,
Cormier J-M,
Carlet J,
Cavaillon J-M:
Influence of surgery on in-vitro cytokine production by human monocytes.
Cytokine
4:576,
1992[Medline]
[Order article via Infotrieve]
27.
Jahns G,
Haeffner-Cavaillon N,
Nydegger UE,
Kazatchkine MD:
Complement activation and cytokine production as consequences of immunological bioincompatibility of extracorporeal circuits.
Clin Materials
14:303,
1993
28.
Bone RC,
Fisher CJ,
Clemmer TP,
Slotman GJ,
Metz CA,
Balk RA:
Sepsis syndrome: a valid clinical entity.
Crit Care Med
17:389,
1989[Medline]
[Order article via Infotrieve]
29.
May CD,
Lyman M,
Alberto R,
Cheng J:
Procedures for immunochemical study of histamine release from leukocytes with small volume of blood.
J Allerg
46:12,
1970
30.
Marty C,
Misset B,
Tamion F,
Fitting C,
Carlet J,
Cavaillon J-M:
Circulating interleukin-8 concentrations in patients with multiple organ failure of septic and nonseptic origin.
Crit Care Med
22:673,
1994[Medline]
[Order article via Infotrieve]
31.
Nicoletti I,
Migliorati G,
Pagliacci MC,
Grignani F,
Riccardi C:
A rapid and simple method for measuring thymocyte apoptosis by propidium iodide staining and flow cytometry.
J Immunol Methods
139:271,
1991[Medline]
[Order article via Infotrieve]
32.
Marie C,
Fitting C,
Cheval C,
Losser MR,
Carlet J,
Payen D,
Foster K,
Cavaillon J-M:
Presence of high levels of leukocyte-associated interleukin-8 upon cell activation and in patients with sepsis syndrome.
Infect Immun
65:865,
1997[Abstract]
33.
Hachiya O,
Takeda Y,
Miyata H,
Watanabe H,
Yamashiat T,
Sendo F:
Inhibition by bacterial lipopolysaccharide of spontaneous and TNF induced human neutrophil apoptosis in vitro.
Microbiol Immunol
39:715,
1995[Medline]
[Order article via Infotrieve]
34.
Biffl WL,
Moore EE,
Moore FA,
Barnett CC:
Interleukin-6 suppression of neutrophil apoptosis is neutrophil concentration dependent.
J Leuk Biol
58:582,
1995[Abstract]
35.
Cavaillon JM,
Pitton C,
Fitting C:
Endotoxin tolerance is not a LPS-specific phenomenon: Partial mimicry with IL-1, IL-10 and TGF.
J Endotoxin Res
1:21,
1994
36.
Kalfin RE,
Engelman RM,
Rousou JA,
Flack JE,
Deaton DW,
Kreutzer DL,
Das DK:
Induction of interleukin-8 expression during cardiopulmonary bypass.
Circulation
88:401,
1993
37.
Sabroe I,
Williams TJ,
Hebert CA,
Collins PD:
Chemoattractant cross-desensitization of the human neutrophil IL-8 receptor involves receptor internalization and differential receptor subtype regulation.
J Immunol
158:1361,
1997[Abstract]
38.
Samanta AK,
Oppenheim JJ,
Matsushima K:
Interleukin-8 dynamically regulates its own receptor expression on human neutrophils.
J Biol Chem
265:183,
1990[Abstract/Free Full Text]
39.
Randow F,
Syrbe U,
Meisel C,
Krausch D,
Zuckermann H,
Platzer C,
Volk H:
Mechanism of endotoxin desensitization: Involvement of interleukin 10 and transforming growth factor .
J Exp Med
181:1887,
1995[Abstract/Free Full Text]
40.
Ertel W,
Kremer J,
Kenney J,
Steckholzer U,
Jarrar D,
Trentz O,
Shildberg F:
Down-regulation of proinflammatory cytokine release in whole blood from septic patients.
Blood
85:1341,
1995[Abstract/Free Full Text]
41. Hallett MB, Lloyds D: Neutrophil priming: The cellular signals
that say "amber" but not "green." Immunol Today 16:264, 1995
42.
Tschaikowsky K,
Sittl R,
Braun GG,
Hering W,
Rügheimer E:
Increased fMet-leu-phe receptor expression and altered superoxide of neutrophil granulocytes in septic and posttraumatic patients.
Clin Invest
72:18,
1993[Medline]
[Order article via Infotrieve]
43.
Rosenbaum JT,
Hartiala KT,
Webster RO,
Howes EL,
Goldstein IM:
Antiinflammatory effects of endotoxin. Inhibition of rabbit polymorphonuclear leukocytes responses to complement (C5)-derived peptides in vivo and in vitro.
Am J Pathol
113:291,
1983[Abstract]
44.
Territo MC,
Golde DW:
Granulocyte function in experimental human endotoxemia.
Blood
47:539,
1976[Abstract/Free Full Text]
45. Hirano S: Migration response of PMN after intraperitoneal and
intratracheal administration of lipopolysaccharide. Am J Physiol (Lung
Cell Mol Physiol) 270:L836, 1996
46.
Parsons PE,
Gillespie MMK,
Moore EE,
Moore FA,
Worthen GS:
Neutrophil response to endotoxin in the adult respiratory distress syndrome: Role of CD14.
Am J Respir Cell Mol Biol
13:152,
1995[Abstract]
47.
Nahum A,
Chamberlin W,
Sznajder JI,
Hegarty M:
Differential activation of mixed venous and arterial neutrophils in patients with sepsis syndrome and acute lung injury.
Am Rev Respir Dis
143:1083,
1991[Medline]
[Order article via Infotrieve]
48.
Chollet-Martin S,
Montravers P,
Gibert C,
Elbim C,
Desmonts JM,
Fagon JY,
Gougerot-Pocidalo MA:
Subpopulation of hyperresponsive polymorphonuclear neutrophils in patients with adult respiratory distress syndrome.
Am Rev Respir Dis
146:990,
1992[Medline]
[Order article via Infotrieve]
49.
Naldini A,
Borrelli E,
Cesari S,
Giomarelli P,
Toscano M:
In vitro cytokine production and T-cell proliferation in patients undergoing cardiopulmonary by-pass.
Cytokine
7:165,
1995[Medline]
[Order article via Infotrieve]
50.
Schinkel C,
Zimmer S,
Kremer JP,
Walz A,
Rordorf-Adam C,
Henckel von Donnersmarck G,
Faist E:
Comparative analysis of transcription and protein release of the inflammatory cytokines interleukin-1 beta (IL-1 beta) and interleukin-8 (IL-8) following major burn and mechanical trauma.
Shock
4:241,
1995[Medline]
[Order article via Infotrieve]
51.
O'Sullivan ST,
Lederer JA,
Horgan AF,
Chin DHL,
Mannick JA,
Rodrick ML:
Major injury leads to predominance of the T helper-2 lymphocyte phenotype and diminished interleukin-12 production associated with decreased resistance to infection.
Ann Surg
222:482,
1995[Medline]
[Order article via Infotrieve]
52.
Wood J,
Rodrick M,
O'Mahony J,
Palder S,
Saporoschetz I,
D'Eon P,
Mannick J:
Inadequate interleukin 2 production. A fundamental immunological deficiency in patients with major burns.
Ann Surg
200:311,
1984[Medline]
[Order article via Infotrieve]
53.
Ertel W,
Morrison M,
Ayala A,
Dean R,
Chaudry I:
Interferon- attenuates hemorrhage-induced suppression of macrophage and splenocyte functions and decreases susceptibility to sepsis.
Surgery
111:177,
1992[Medline]
[Order article via Infotrieve]
54.
Hill GE,
Pohorecki R,
Alonso A,
Rennard SI,
Robbins RA:
Aprotinin reduces interleukin-8 production and lung neutrophil accumulation after cardiopulmonary bypass.
Anesth Analg
83:696,
1996[Abstract]
55.
Kawamura T,
Wakusawa R,
Okada K,
Inada S:
Elevation of cytokines during open heart surgery with cardiopulmonary bypass: Participation of interleukin 8 and 6 in reperfusion injury.
Can J Anaesth
40:1016,
1993[Medline]
[Order article via Infotrieve]
56.
Jorens PG,
De Jongh R,
De Backer W,
Van Damme J,
Van Overveld F,
Bossaert L,
Walter P,
Herman AG,
Rampart M:
Interleukin-8 production in patients undergoing cardiopulmonary bypass. The influence of pretreatment with methylprednisolone.
Am Rev Respir Dis
148:890,
1993[Medline]
[Order article via Infotrieve]
57.
Jansen NJG,
van Oeverren W,
Gu YJ,
van Vliet MH,
Eijsman L,
Wildevuur CRH:
Endotoxin release and tumor necrosis factor formation during cardiopulmonary bypass.
Ann Thorac Surg
54:744,
1992[Abstract]
58.
Dehoux M,
Philip I,
Chollet-Martin S,
Boutten A,
Hvass U,
Desmonts JM,
Durand G:
Early production of interleukin-10 during normothermic cardiopulmonary bypass.
J Thorac Cardiovasc Surg
110:286,
1995[Free Full Text]
59.
Borrelli E,
Giomarelli P,
Naldini A,
Luzzi E,
Silvestri S,
Gardinali M,
Bocci V:
Plasma levels of immunosuppressive mediators during cardiopulmonary bypass.
Mediat Inflamm
5:51,
1996
60.
Haeffner-Cavaillon N,
Rousselier N,
Pnozio O,
Carreno M,
Laude M,
Carpentier A,
Kazatchkine M:
Induction of Interleukin-1 production in patients undergoing cardiopulmonary bypass.
J Thorac Cardiovasc Surg
98:1100,
1989[Abstract]
61.
Watarida S,
Mori A,
Onoe M,
Tabat R,
Shiraishi S,
Sugita T,
Nojima T,
Nakajima Y,
Matsuno S:
A clinical study on the effects of pulsatile cardiopulmonary bypass on the blood endotoxin levels.
J Thorac Cardiovasc Surg
108:620,
1994[Abstract/Free Full Text]
62.
Takasuka N,
Tokunaga T,
Akagawa KS:
Preexposure of macrophages to low doses of lipopolysaccharide inhibits the expression of tumor necrosis factor- mRNA but not of IL-1 mRNA.
J Immunol
146:3824,
1991[Abstract]
63.
Marchant A,
Devière J,
Byl B,
De Groote D,
Vincent J,
Goldman M:
Interleukin-10 production during septicaemia.
Lancet
343:707,
1994[Medline]
[Order article via Infotrieve]
64.
Guthrie LA,
McPhail LC,
Henson PM,
Johnston RB:
Priming of neutrophils for enhanced release of oxygen metabolites by bacterial lipopolysaccharide.
J Exp Med
160:1656,
1984[Abstract/Free Full Text]
65.
Surette ME,
Palmantier R,
Gosselin J,
Borgeat P:
Lipopolysaccharide prime whole human blood and isolated neutrophils for the increased synthesis of 5-lipoxygenase products by enhancing arachidonic acid availability: Involvement of the CD14 antigen.
J Exp Med
178:1347,
1993[Abstract/Free Full Text]
66.
Schleiffenbaum B,
Fehr J:
The tumor necrosis factor receptor and human neutrophil function. Deactivation and cross-deactivation of TNF-induced neutrophil responses by receptor down-regulation.
J Clin Invest
86:184,
1990
 CiteULike  Connotea  Del.icio.us  Digg  Reddit  Technorati What's this?
This article has been cited by other articles:
|
|
|
|
 
N. Tamassia, F. Calzetti, N. Menestrina, M. Rossato, F. Bazzoni, L. Gottin, and M. A. Cassatella
Circulating neutrophils of septic patients constitutively express IL-10R1 and are promptly responsive to IL-10
Int. Immunol.,
April 1, 2008;
20(4):
535 - 541.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J.-M. Cavaillon and D. Annane
Invited review: Compartmentalization of the inflammatory response in sepsis and SIRS
Innate Immunity,
June 1, 2006;
12(3):
151 - 170.
[Abstract]
[PDF]
|
|
|
|
|
|
|
L. C. Parker, E. C. Jones, L. R. Prince, S. K. Dower, M. K. B. Whyte, and I. Sabroe
Endotoxin tolerance induces selective alterations in neutrophil function
J. Leukoc. Biol.,
December 1, 2005;
78(6):
1301 - 1305.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J.-M. Cavaillon, C. Adrie, C. Fitting, and M. Adib-Conquy
Reprogramming of circulatory cells in sepsis and SIRS
Innate Immunity,
October 1, 2005;
11(5):
311 - 320.
[Abstract]
[PDF]
|
|
|
|
|
|
|
M. Adib-Conquy, P. Moine, K. Asehnoune, A. Edouard, T. Espevik, K. Miyake, C. Werts, and J.-M. Cavaillon
Toll-like Receptor-mediated Tumor Necrosis Factor and Interleukin-10 Production Differ during Systemic Inflammation
Am. J. Respir. Crit. Care Med.,
July 15, 2003;
168(2):
158 - 164.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
H. Corvol, C. Fitting, K. Chadelat, J. Jacquot, O. Tabary, M. Boule, J.-M. Cavaillon, and A. Clement
Distinct cytokine production by lung and blood neutrophils from children with cystic fibrosis
Am J Physiol Lung Cell Mol Physiol,
June 1, 2003;
284(6):
L997 - L1003.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J.-M. Cavaillon, C. Adrie, C. Fitting, and M. Adib-Conquy
Endotoxin tolerance: is there a clinical relevance?
Innate Immunity,
April 1, 2003;
9(2):
101 - 107.
[Abstract]
[PDF]
|
|
|
|
|
|
|
C. Adrie, M. Adib-Conquy, I. Laurent, M. Monchi, C. Vinsonneau, C. Fitting, F. Fraisse, A. T. Dinh-Xuan, P. Carli, C. Spaulding, et al.
Successful Cardiopulmonary Resuscitation After Cardiac Arrest as a "Sepsis-Like" Syndrome
Circulation,
July 30, 2002;
106(5):
562 - 568.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A. S. Cross
Invited review: Endotoxin tolerance -- current concepts in historical perspective
Innate Immunity,
April 1, 2002;
8(2):
83 - 98.
[PDF]
|
|
|
|
|
|
|
J. Zweigner, H.-J. Gramm, O. C. Singer, K. Wegscheider, and R. R. Schumann
High concentrations of lipopolysaccharide-binding protein in serum of patients with severe sepsis or septic shock inhibit the lipopolysaccharide response in human monocytes
Blood,
December 15, 2001;
98(13):
3800 - 3808.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J.-M. Cavaillon, M. Adib-Conquy, I. Cloez-Tayarani, and C. Fitting
Review: Immunodepression in sepsis and SIRS assessed by ex vivo cytokine production is not a generalized phenomenon: a review
Innate Immunity,
April 1, 2001;
7(2):
85 - 93.
[Abstract]
[PDF]
|
|
|
|
|
|
|
D. P. Olszyna, E. De Jonge, P. E. P. Dekkers, S. J. H. van Deventer, and T. van der Poll
Induction of Cell-Associated Chemokines after Endotoxin Administration to Healthy Humans
Infect. Immun.,
April 1, 2001;
69(4):
2736 - 2738.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
R. S. MUNFORD and J. PUGIN
Normal Responses to Injury Prevent Systemic Inflammation and Can Be Immunosuppressive
Am. J. Respir. Crit. Care Med.,
February 1, 2001;
163(2):
316 - 321.
[Full Text]
|
|
|
|
|
|
|
M.T. MASCELLINO, G. DELOGU, M.R. PELAIA, R. PONZO, R. PARRINELLO, and A. GIARDINA
Reduced bactericidal activity against Staphylococcus aureus and Pseudomonas aeruginosa of blood neutrophils from patients with early adult respiratory distress syndrome
J. Med. Microbiol.,
January 1, 2001;
50(1):
49 - 54.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. ADIB-CONQUY, C. ADRIE, P. MOINE, K. ASEHNOUNE, C. FITTING, M. R. PINSKY, J.-F. DHAINAUT, and J.-M. CAVAILLON
NF-kappa B Expression in Mononuclear Cells of Patients with Sepsis Resembles That Observed in Lipopolysaccharide Tolerance
Am. J. Respir. Crit. Care Med.,
November 1, 2000;
162(5):
1877 - 1883.
[Abstract]
[Full Text]
|
|
|
|
|
|
|
N. Rayhane, C. Fitting, O. Lortholary, F. Dromer, and J.-M. Cavaillon
Administration of Endotoxin Associated with Lipopolysaccharide Tolerance Protects Mice against Fungal Infection
Infect. Immun.,
June 1, 2000;
68(6):
3748 - 3753.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
P. Massoudy, S. Zahler, P. Tassani, B. F. Becker, J. A. Richter, M. Pfauder, R. Lange, and H. Meisner
Reduction of pro-inflammatory cytokine levels and cellular adhesion in CABG procedures with separated pulmonary and systemic extracorporeal circulation without an oxygenator
Eur. J. Cardiothorac. Surg.,
June 1, 2000;
17(6):
729 - 736.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
N. Kotani, H. Hashimoto, D. I. Sessler, M. Muraoka, J.-S. Wang, M. F. O'Connor, and A. Matsuki
Cardiopulmonary Bypass Produces Greater Pulmonary than Systemic Proinflammatory Cytokines
Anesth. Analg.,
May 1, 2000;
90(5):
1039 - 1045.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
F. N. Lauw, T. ten Hove, P. E. P. Dekkers, E. de Jonge, S. J. H. van Deventer, and T. van der Poll
Reduced Th1, but Not Th2, Cytokine Production by Lymphocytes after In Vivo Exposure of Healthy Subjects to Endotoxin
Infect. Immun.,
March 1, 2000;
68(3):
1014 - 1018.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
P. Massoudy, S. Zahler, T. Freyholdt, R. Henze, A. Barankay, B. F. Becker, S. L. Braun, and H. Meisner
SODIUM NITROPRUSSIDE IN PATIENTS WITH COMPROMISED LEFT VENTRICULAR FUNCTION UNDERGOING CORONARY BYPASS: REDUCTION OF CARDIAC PROINFLAMMATORY SUBSTANCES
J. Thorac. Cardiovasc. Surg.,
March 1, 2000;
119(3):
566 - 574.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J.-M. Cavaillon, M. Adib-Conquy, C. Marie, and C. Fitting
Hyporesponsiveness in leukocytes in sepsis: in vitro models reveal paradoxical effects of IL-10
Innate Immunity,
February 1, 1999;
5(1-2):
81 - 85.
[Abstract]
[PDF]
|
|
|
|
|
|
|
M. Weiss, L. L. Moldawer, and E. M. Schneider
Granulocyte Colony-Stimulating Factor to Prevent the Progression of Systemic Nonresponsiveness in Systemic Inflammatory Response Syndrome and Sepsis
Blood,
January 15, 1999;
93(2):
425 - 439.
[Full Text]
[PDF]
|
|
|
|
|
Abstract
Introduction
Abstract