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B
into Human Macrophages: Lipopolysaccharide-Induced, But Not Zymosan-Induced, Proinflammatory Cytokines Are Inhibited, But IL-10 Is Nuclear Factor-B Independent1
Kennedy Institute of Rheumatology, Hammersmith, London, United Kingdom
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Abstract |
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 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
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production in rheumatoid
arthritis joint cells, chiefly from macrophages, is 75% blocked
by adenoviral transfer of I
B. In this report we use the same
adenovirus to investigate whether the production of a number of
proinflammatory cytokines (e.g., TNF-, IL-1ß, IL-6, and IL-8) from
human macrophages depends on NF-B. While the cytokine response to
certain inducers, such as LPS, PMA, and UV light, is blocked by
overexpression of IB, the response to zymosan is not. In
contrast, anti-inflammatory mediators (IL-10 and IL-1 receptor
antagonist) induced by LPS are only marginally inhibited by IB
excess. These studies demonstrate several new points about macrophage
cytokine production. First, there is heterogeneity of mechanisms
regulating both the proinflammatory and anti-inflammatory cytokines
within populations of a single cell type. In addition, the results
confirm the utility of the adenoviral technique for functional analysis
of cytokine induction. The results also confirm that there are
autocrine and paracrine interactions regulating cytokine synthesis
within a single cell type. The selectivity of NF-B blockade for
proinflammatory but not anti-inflammatory mediators indicates that
in macrophages, NF-B may be a good target for the treatment of
chronic inflammatory diseases. |
Introduction |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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and, consequently,
other proinflammatory cytokines 1, 2 . The most definitive evidence in
humans comes from the success of clinical trials in these diseases,
using strategies of blocking TNF- through mAbs (e.g.,
Remicade or Infliximab; Centocor, Malvern, PA) 3 or soluble
TNF receptors (Enbrel; Immunex, Seattle, WA) 4 . The
anti-inflammatory mediators and cytokines, such as IL-10, the
soluble TNF receptors, and the IL-1 receptor antagonist
(IL-1ra),3 are also
up-regulated in rheumatoid synovial tissue, but they are only partly
able to antagonize the effects of the proinflammatory mediators 1 .
The successful clinical trials of anti-TNF- Ab in rheumatoid
arthritis 3, 5 , reviewed in Reference 2, and Crohn’s disease 6
have prompted considerable interest in alternative strategies to
inhibit the production of TNF-. Because macrophages are the major
producers of TNF- and other proinflammatory cytokines in the
rheumatoid joint 1 , as well as of the anti-inflammatory
cytokines, understanding the signaling pathways involved in the
induction of these mediators is of major importance for developing
novel therapeutic strategies in chronic inflammatory diseases.
In a previous paper 7 , we described a novel technique to achieve
efficient, virtually 100%, adenoviral gene transfer into human
macrophages, subsequent to up-regulation of integrins. This technique
was used to effect adenoviral transfer of the IB molecule into
human macrophages, as previously reported in endothelial cells 8 .
Massive overexpression of IB was achieved, with consequent
inhibition of NF-B activity. It was observed that the LPS-induced
expression of TNF- in human macrophages was potently inhibited by
the blocking NF-B. This was in contrast to previous studies in human
cells, which did not indicate a role for NF-B in TNF expression 47, 48 . However, unlike our studies, these had been performed in
transformed cell lines mainly of lymphoid origin, which may not reflect
the situation in normal macrophages.
In this paper, the effect of NF-B down-regulation on a spectrum of
proinflammatory cytokines, namely IL-1ß, IL-6, and IL-8, and
anti-inflammatory molecules IL-10, IL-1ra, and the p55 and p75
soluble TNF-receptors is studied. The aim was to resolve some of the
conflicting evidence concerning macrophage production of cytokines.
While there is reasonably good evidence that TNF- 9, 10 , IL-1ß
11, 12 , and IL-6 13 can be regulated by NF-B in various cell
types, there are few data concerning the other cytokines of interest.
The lack of B binding sites in the human IL-10 promoter made it
unlikely that IL-10 was under direct NF-B control 14 .
Another aim of this study was to use the IB adenovirus as a tool to
determine whether different macrophage activators, e.g., LPS, PMA, UV
light, and zymosan, were dependent on NF-B. In the RAW 264.7
macrophage-like cell line, the UV-induced TNF- response has been
proposed not to involve NF-B activation, since it was unaffected by
mutation of all four B sites within the TNF- promoter 15 . With
regard to the activation of monocytic cells induced by PMA, results
disagree: some studies 16, 17 suggest that NF-B is involved, and
others 18 do not. Recently, in a model of THP-1 cells modified
through stable retroviral gene transfer of IB, PMA-induced IL-1,
IL-6, and IL-8 were unaffected 19 . Zymosan particles are yeast cell
derivatives that induce cytokines and are used to simulate
receptor-mediated cell stimulation, probably in the ß-glucan receptor
20 . Both in mouse bone marrow-derived macrophages 21 and in human
monocytes 22 , binding of zymosan induces rapid tyrosine
phosphorylation of a number of protein substrates. In human monocytes,
zymosan induces association of the p53–56lyn tyrosine kinases
and the cytoskeleton 23 . The tyrosine kinase inhibitor genistein
proved to be a potent inhibitor of zymosan-induced eicosanoid formation
in mouse peritoneal macrophages 24 . There is some doubt, however, as
to whether zymosan acts via NF-B. In rat liver macrophages, zymosan
is incapable of activating NFB 18 , although it does activate the
transcription factor activator protein-1 both in rat liver macrophages
18 and in the U937 human monocytic cell line 25 .
The adenoviral infection technique 7 has enabled us to address many
of these issues regarding the NF-B dependence or independence of the
signal transduction pathways utilized by these various stimuli in human
macrophages.
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Materials and Methods |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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Single-donor plateletphoresis residues were purchased from North London Blood Transfusion Service (Colindale, U.K.). Mononuclear cells were isolated by Ficoll-Hypaque centrifugation preceding monocyte separation in a Beckman Instruments (Torrence, CA) JEL elutriator. Monocyte purity was assessed by flow cytometry and was routinely >90%.
Adenoviral vectors
Recombinant, replication-deficient adenoviral vectors encoding
Escherichia coli ß-galactosidase or having no insert
(Adv0) were generously provided by Drs. A. Byrnes and M. Wood (Oxford,
U.K.). An adenovirus encoding porcine IB (AdvIB) with a CMV
promoter and a nuclear localization sequence was generously provided by
Dr. R. de Martin (Vienna, Austria). Viruses were propagated in the 293
human embryonic kidney cell line and purified by ultracentrifugation
through two cesium chloride gradients. The titers of viral stocks were
determined through a plaque assay on 293 cells, as described 26 .
Infection techniques
The elutriated human monocytes were incubated at 
2 x
106/ml in RPMI 1640 with 25 mM HEPES and 2 mM
L-glutamine supplemented with 5% (v/v) heat-inactivated
FCS and 10 U/ml penicillin/streptomycin. To optimize infection,
purified human monocytes were pretreated with macrophage CSF (100
ng/ml; obtained from Genetics Institute, Boston, MA) for 48 h to
allow up-regulation of integrin Vß5, which
has previously been shown to be essential for adenovirus infection of
monocytes 27 . The cells were then replated on 100-mm petri dishes and
infected for 2 h with a multiplicity of infection (MOI) of between
10:1 and 120:1 (in most experiments, 20:1, 40:1, or 80:1 was used) of
either AdvIB or Adv0, in serum-free RPMI 1640. Cells were then
incubated in RPMI 1640 supplemented as above for 48 h to allow for
significant overexpression of IB, as assessed 7 . During the
changes of medium involved, nonadherent cells were discarded, resulting
in a further purification of monocyte-derived macrophages.
Cytokine analysis
For Northern blot analysis experiments, cells were replated at 5–10 x 106 cells per 100-mm petri dish and stimulated with LPS (10 ng/ml), PMA (10 nM), zymosan (30 µg/ml), ionomycin (1 µM), or UV irradiation (2000 J). After 4 h, cells were harvested, and mRNA was extracted and subjected to Northern blot analysis as in Reference 28.
In the assays for cytokine production, cells were replated at 5 x
105 cells per well on a 96-well dish and stimulated as
above for 4 or 16 h. Supernatants were analyzed for
TNF- 29 , IL-1ß, IL-6 and IL-8 30 , IL-10 31, 32 , IL-1ra, and
the p55 and p75 soluble TNF receptors 33 by ELISA.
The proteosome inhibitor
benzyloxycarbonyl-Ile-Glu(O-tert-butyl)-Ala-leucinal
(PSI) was obtained from Calbiochem (Nottingham, U.K.).
Electrophoretic mobility shift assay
Nuclear extracts were prepared and 20 µg of protein was
analyzed for NF-B activity as previously described 34 .
Statistical methods
All statistical testing was performed using a paired comparison, one-sided Student’s t test, except when a Scheffé test of multiple comparisons was used as indicated 35 .
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Results |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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LPS is capable of inducing all the cytokines (TNF-, IL-1ß,
IL-6, and IL-8) and inhibitors (IL-10, IL-1ra, and the soluble TNF
receptors) assayed in this study (Table I
; results are means of 4–11
experiments) each using blood from different donors. The induction of
TNF- mRNA by zymosan has previously been reported to be far weaker
than LPS-induced TNF- mRNA in mouse peritoneal macrophages 36 , but
the TNF- response to zymosan in macrophage CSF-treated human
monocytes was equal to that of LPS (Table I).
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), whereas PMA
could induce IL-1ra and p75 soluble TNF receptor.
Does IB overexpression inhibit LPS-induced proinflammatory
cytokines?
We previously observed that infection at an MOI of 20–80:1 with
the IB adenovirus produced high levels of IB expression.
This resulted in a potent inhibition of LPS-driven TNF- induction in
human macrophages by blocking NFB function 7 . This was not due to
loss of cells through apoptosis or other causes of cell death 7 . The
induction of IL-1ß and IL-8 by LPS also appears to be strongly
NF-B dependent (Fig. 1, A
and B), and there is potent inhibition of both of these
cytokines already at 20:1 of AdvIB. Results using supernatants
harvested after 4 h of incubation were similar to those after
16 h of incubation (not shown). In contrast to the other cytokines
studied here, there was a potentiation of IL-6 production of
20–30% in Adv0-infected cells, with LPS as well as with other
stimuli (Fig. 1C), although adenovirus infection alone had
no effect. Nevertheless, IL-6 expression was strongly inhibited by
infection with AdvIB in human macrophages (Fig. 1C).
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can induce the synthesis of other cytokines, e.g.,
IL-1ß, IL-6, and IL-8, the potent inhibition by AdvIB might be
secondary to inhibition of TNF-. Culturing with exogenous TNF-
partly abrogated the effect of IB overexpression on IL-1ß
production (Fig. 2A),
indicating at least a partial dependence on TNF- (there was just a
20% inhibition in the presence of 100 ng/ml TNF-, compared with
62% inhibition with no cytokine; difference was significant
(p < 0.005) between these two comparisons
using a Scheffé multiple-comparisons test). However, similar
experiments showed no major role for TNF- in LPS-induced expression
of IL-6 and IL-8 (results not shown).
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-driven IL-1ß response might be less
NF-B dependent than the IL-1ß response induced by LPS. One
difficulty in doing this was the fact that TNF- is a weaker inducer
of IL-1ß in our system than LPS (Table I). However, the
IB-induced inhibition of TNF--induced IL-1ß was relatively
less potent than that seen with TNF--induced IL-6 (Fig. 2, B and C), as expected from the restoration of
IL-1ß synthesis by TNF- in cells infected with AdvIB.
Does IB overexpression affect LPS-induced
anti-inflammatory cytokines?
The major anti-inflammatory cytokine produced by macrophages
is IL-10 37, 38 . In LPS-stimulated cells, there was gradual
inhibition with increasing virus titers, but statistically significant
inhibition (p < 0.05) was only noted at 60:1
or 80:1 of AdvIB (Fig. 3A). This inhibition was still
quite modest (30% at most), and since the human IL-10 promoter lacks
B sites 14 , we investigated whether the inhibitory effects of
AdvIB infection were indirect, occurring via its effects on
proinflammatory cytokines that are known to influence IL-10 expression
32, 39, 40 . A combination of TNF- and LPS considerably potentiated
the IL-10 response and partially abrogated the inhibition by
AdvIB (Fig. 3B). In contrast, IL-1 failed to have any
significant effect (results not shown).
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30%) by IB
overexpression (Fig. 4A). This
response was only modestly affected by adding back IL-1 and TNF-
(Fig. 4B). In human macrophages, LPS induces the production
of the p75 soluble TNF receptor and to a lesser extent the p55 soluble
TNF receptor (Table I). Infection with 40:1 of AdvIB was observed
to significantly (p < 0.001) inhibit the
LPS-induced production of both of these soluble receptors (Fig. 5); this response was unaffected by
adding back IL-1 or TNF- (results not shown).
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B overexpression inhibit proinflammatory cytokines
induced by PMA or UV light?
The induction of TNF- by PMA is very potently (90%)
inhibited by IB overexpression, even more strongly than in
LPS-stimulated (80%) 7 or even UV light-stimulated cells
(80%) (Table II). Again, results
using supernatants harvested after 4 h of incubation were similar
to those using 16 h of incubation (data not shown). A series of
Northern blot analysis experiments demonstrated that, as for
LPS-induced TNF- 7 , the TNF- mRNA expression in response to PMA
and UV light was ablated by IB overexpression (Fig. 6, A and B).
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, the induction of IL-1ß and IL-6 by PMA or UV
light was NF-B dependent (Table II). Once again, the inhibition of
IL-1ß and IL-6 production by the AdvIB infection appears to be
more potent in PMA-treated cells than in cells treated with LPS or UV
light.
There was also significant (p < 0.001)
inhibition (50–60%) of the IL-8 response when cells infected with
AdvIB were stimulated with PMA or UV light (Table II). Ionomycin,
a stimulus that did not induce significant amounts of the other
cytokines of interest, induced a discernible IL-8 response, which was
also inhibited by IB overexpression (data not shown).
PMA did not induce detectable amounts of IL-10, but it induced a
detectable p75 soluble TNF receptor response, which was significantly
(p < 0.001) inhibited (75% at 40:1 of
AdvIB) by IB overexpression, as was the PMA-induced
production of IL-1ra (40% inhibition at 40:1 of AdvIB;
p < 0.005).
Does IB overexpression affect zymosan-induced cytokines?
In contrast to the prior stimuli when the macrophages were
activated with zymosan, infection with AdvIB had no effect
whatsoever on TNF- protein (Table II) or mRNA expression (Fig. 6C), even at MOI of 80:1. The induction of IL-1ß and
IL-8 by zymosan was also unaffected by the IB overexpression
(Table II), but there was a small (10–15% compared with
uninfected cells) and statistically nonsignificant inhibition of IL-6.
Zymosan-induced IL-10, IL-1ra (Table II), and the soluble TNF
receptors (not shown) were also refractory to inhibition by
IB overexpression.
The independence from NF-B of zymosan-induced TNF was further
emphasized by studies with the proteosome inhibitor PSI. Inhibition of
proteosome function inhibits IB degradation, thus preventing
NF-B nuclear translocation 41, 42 . PSI was very effective in
blocking LPS-induced TNF production, but it did not affect the response
to zymosan (Fig. 7).
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B,
the activation of this transcription factor by zymosan (30 µg/ml) was
slower (detectable only after 60 min) and much weaker (Fig. 8). Increasing the zymosan concentration
did not enhance its effect on NF-B (not shown). The activation of
NF-B by either stimulus was inhibited by IB overexpression
(Fig. 8). Although a 60-min incubation with TNF- (1–10 ng/ml)
activated NF-B weakly, lower concentrations did not, and the amount
of TNF- produced after a 60-min incubation with zymosan is <100
pg/ml (not shown), arguing against a feedback loop of this kind being
involved.
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Discussion |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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B
effectively inhibits NF-B activity in human macrophages, mainly
through the overexpression of IB inhibiting nuclear translocation
of the p65/p50 subunits of NF-B. This blocked LPS-induced TNF-
both at the mRNA and the protein levels. Even more interesting, it was
found that the endogenous production of TNF- from rheumatoid
synovial mononuclear cell cultures was also inhibited 7 .
The fact that LPS-induced TNF-, IL-1ß, and IL-6 were all NF-B
dependent cytokines could be expected from the majority of data from
murine cells and monocyte/macrophage cell lines 9, 10 . Our results on
LPS-induced cytokines also agree well with the results of Makarov et
al. 19 on LPS-induced IL-1ß, IL-6, and IL-8 in monocytic THP-1
cells stably modified through retroviral gene transfer of IB. Among
the LPS-induced, proinflammatory cytokines studied here, IL-6 was most
potently inhibited (>85%) by overexpression of IB, whereas
there was always some residual production of TNF- or IL-1ß even in
LPS-stimulated cells infected with high titers (120:1) of AdvIB
(not shown). This may reflect a certain amount of preformed cytokine
mRNA, but this could not be demonstrated in unstimulated cells (data
not shown), and furthermore LPS 7 , PMA-induced or UV-induced (Fig. 6)
TNF- mRNA was profoundly down-regulated by IB. However, it
could not be excluded that this residual cytokine production emanated
from the few uninfected cells still present, and work is in progress to
elucidate this question using intracytoplasmic staining for cytokines.
Another nonexclusive hypothesis (discussed in more detail below) is
that TNF- and other proinflammatory cytokines can be induced through
both NF-B-dependent and NF-B-independent pathways.
Our finding that UV light induces a whole spectrum of proinflammatory
cytokines in a NF-B-dependent manner is novel. It is in contrast to
the earlier report that UV-induced TNF- in RAW 264.7 cells does not
involve NF-B 15 . Similarly, the finding that the PMA-induced
induction of TNF- and other proinflammatory cytokines is profoundly
down-regulated by IB overexpression disagrees with several
earlier studies in stably transformed human cell lines 19 . In our
hands, this stimulus was actually the one most strongly dependent on
NF-B, as judged by the percentage of inhibition, reproduced in seven
separate experiments. These discrepancies between results obtained with
human primary cells and those from various transformed cell lines
indicate that, at least in some instances, the latter are questionable
models for studying cytokine cell signaling occurring in primary cells,
as is the case in vivo. In a way, this is not surprising, since there
are interactions between the enzymes and transcription factors of the
cell cycle machinery and the regulation of cytokine genes, e.g., Rb
regulates ets, which is involved in cytokine activation
43 .
With regard to macrophage signal transduction, one of the most
remarkable findings in the present study was that zymosan, although a
very powerful macrophage activator, does not appear to require NF-B
for the induction of either pro- or anti-inflammatory cytokines.
These findings would imply that there are, in human macrophages, both
NF-B-dependent and NF-B-independent pathways of cytokine
induction involved in the induction of TNF- and other
proinflammatory cytokines. Although zymosan does activate NF-B, it
does so more slowly and much less potently than LPS (Fig. 8), and it is
likely that some other transcription factor mediates the
zymosan-induced cytokine production. The modest (15%) inhibition of
zymosan-induced IL-6 observed in cells overexpressing IB is of
questionable significance and may reflect the observation that this
cytokine was the most potently affected by IB overexpression,
irrespective of stimulus (see Table II).
Another finding of importance is that in human macrophages, IL-10 is
under complex control, and in LPS-stimulated cells, it appears to be at
least partially driven via LPS-induced TNF- and IL-1. It is
interesting to note that, even at 40:1 of AdvIB, when LPS-driven
TNF- is abrogated by >60% 7 , IL-10 is still not significantly
inhibited (Fig. 3). At higher virus titers, resulting in even stronger
inhibition of TNF-, there is some effect also on IL-10, but never,
even with 80:1 of the virus, exceeding 30%. This is completely
reversible by adding back TNF-, which implies that LPS-induced IL-10
is partly driven secondarily by TNF-. This finding agrees well with
previous reports 14, 32, 40, 44 and indicates that autocrine
interactions can take place, even in short-term (16-h) cultures such as
these.
Another intriguing finding was that the IB-induced inhibition of
the LPS-induced production of IL-1ß (Fig. 2A), but not the
production of IL-6, IL-8, or the soluble TNF receptors (data not
shown), was also somewhat abrogated when TNF- was restored. This
indicates that TNF-induced IL-1ß is mainly independent of NF-B.
This finding also suggests that IL-1ß is also, although to a lesser
extent than IL-10, driven partly by LPS-induced TNF-. This result
echoes the previous work in rheumatoid arthritis joint cell
cocultures, in which TNF- blockade was found to inhibit the
production of IL-1 45 and subsequently of IL-6, IL-8, IL-10 and
granulocyte-macrophage CSF 30, 46 , which has led to the concept of a
TNF--dependent "cytokine cascade" in inflammatory sites such as
the rheumatoid synovium 1 . The current macrophage cultures are
shorter term than the rheumatoid synovial cultures (16 h vs 5 days),
which may explain why the "cascade" appears more marked in the
latter system.
The dissection out of signaling pathways in normal primary cells is
necessary, as there are differences from cell lines (see above). This
is now possible within human macrophages, from either normal or
pathological specimens, using this adenoviral technique. The model of
human macrophages infected with AdvIB has, from a cytokine point
of view, provided results similar to those from infecting human
synovial cocultures with the same virus 7 (J. Bondeson et al.,
manuscript in preparation). Taken together, the data suggest that
NF-B is an important therapeutic target in chronic inflammatory
diseases, allowing profound down-regulation of macrophage-produced
proinflammatory cytokines while not directly affecting the most
important anti-inflammatory cytokines, IL-10 and IL-1ra. This would
redress the disturbed equilibrium between these mediators 1 .
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Acknowledgments |
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B adenovirus. |
Footnotes |
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2 Address correspondence and reprint requests to Dr. Marc Feldmann, Kennedy Institute of Rheumatology, 1 Aspenlea Road, Hammersmith, London W6 8LH, U.K. E-mail address:
3 Abbreviations used in this paper: IL-1ra, IL-1R antagonist; Adv0, adenovirus with no insert; AdvIB, adenovirus encoding porcine IB; MOI, multiplicity of infection; PSI, benzyloxycarbonyl-Ile-Glu(O-tert-butyl)-Ala-leucinal.
Received for publication August 14, 1998. Accepted for publication November 16, 1998.
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 M. E. Killeen, J. A. Englert, D. B. Stolz, M. Song, Y. Han, R. L. Delude, J. A. Kellum, and M. P. Fink The Phase 2 Enzyme Inducers Ethacrynic Acid, DL-Sulforaphane, and Oltipraz Inhibit Lipopolysaccharide-Induced High-Mobility Group Box 1 Secretion by RAW 264.7 Cells J. Pharmacol. Exp. Ther., March 1, 2006; 316(3): 1070 - 1079. [Abstract] [Full Text] [PDF]
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K. M. Stuhlmeier and C. Pollaschek Adenovirus-mediated Gene Transfer of Mutated I{kappa}B Kinase and I{kappa}B{alpha} Reveal NF-{kappa}B-dependent as Well as NF-{kappa}B-independent Pathways of HAS1 Activation J. Biol. Chem., December 30, 2005; 280(52): 42766 - 42773. [Abstract] [Full Text] [PDF]
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C. C. Taggart, S.-A. Cryan, S. Weldon, A. Gibbons, C. M. Greene, E. Kelly, T. B. Low, S. J. O'Neill, and N. G. McElvaney Secretory leucoprotease inhibitor binds to NF-{kappa}B binding sites in monocytes and inhibits p65 binding J. Exp. Med., December 19, 2005; 202(12): 1659 - 1668. [Abstract] [Full Text] [PDF]
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P. Zhang, M. Martin, S. M. Michalek, and J. Katz Role of Mitogen-Activated Protein Kinases and NF-{kappa}B in the Regulation of Proinflammatory and Anti-Inflammatory Cytokines by Porphyromonas gingivalis Hemagglutinin B Infect. Immun., July 1, 2005; 73(7): 3990 - 3998. [Abstract] [Full Text] [PDF]
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 H. M. Wilson, S. Chettibi, C. Jobin, D. Walbaum, A. J. Rees, and D. C. Kluth Inhibition of Macrophage Nuclear Factor-{kappa}B Leads to a Dominant Anti-Inflammatory Phenotype that Attenuates Glomerular Inflammation in Vivo Am. J. Pathol., July 1, 2005; 167(1): 27 - 37. [Abstract] [Full Text] [PDF]
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Y. Cong, A. Konrad, N. Iqbal, R. D. Hatton, C. T. Weaver, and C. O. Elson Generation of Antigen-Specific, Foxp3-Expressing CD4+ Regulatory T Cells by Inhibition of APC Proteosome Function J. Immunol., March 1, 2005; 174(5): 2787 - 2795. [Abstract] [Full Text] [PDF]
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 W. L. W. Chang, N. Baumgarth, D. Yu, and P. A. Barry Human Cytomegalovirus-Encoded Interleukin-10 Homolog Inhibits Maturation of Dendritic Cells and Alters Their Functionality J. Virol., August 15, 2004; 78(16): 8720 - 8731. [Abstract] [Full Text] [PDF]
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 R. L. Chelvarajan, S. M. Collins, I. E. Doubinskaia, S. Goes, J. Van Willigen, D. Flanagan, W. J. S. de Villiers, J. S. Bryson, and S. Bondada Defective macrophage function in neonates and its impact on unresponsiveness of neonates to polysaccharide antigens J. Leukoc. Biol., June 1, 2004; 75(6): 982 - 994. [Abstract] [Full Text] [PDF]
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 C. Monaco, E. Andreakos, S. Kiriakidis, C. Mauri, C. Bicknell, B. Foxwell, N. Cheshire, E. Paleolog, and M. Feldmann Canonical pathway of nuclear factor {kappa}B activation selectively regulates proinflammatory and prothrombotic responses in human atherosclerosis PNAS, April 13, 2004; 101(15): 5634 - 5639. [Abstract] [Full Text] [PDF]
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W. Ma, K. Gee, W. Lim, K. Chambers, J. B. Angel, M. Kozlowski, and A. Kumar Dexamethasone Inhibits IL-12p40 Production in Lipopolysaccharide-Stimulated Human Monocytic Cells by Down-Regulating the Activity of c-Jun N-Terminal Kinase, the Activation Protein-1, and NF-{kappa}B Transcription Factors J. Immunol., January 1, 2004; 172(1): 318 - 330. [Abstract] [Full Text] [PDF]
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L. Ziegler-Heitbrock, M. Lotzerich, A. Schaefer, T. Werner, M. Frankenberger, and E. Benkhart IFN-{alpha} Induces the Human IL-10 Gene by Recruiting Both IFN Regulatory Factor 1 and Stat3 J. Immunol., July 1, 2003; 171(1): 285 - 290. [Abstract] [Full Text] [PDF]
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 T. Ronni, V. Agarwal, M. Haykinson, M. E. Haberland, G. Cheng, and S. T. Smale Common Interaction Surfaces of the Toll-Like Receptor 4 Cytoplasmic Domain Stimulate Multiple Nuclear Targets Mol. Cell. Biol., April 1, 2003; 23(7): 2543 - 2555. [Abstract] [Full Text] [PDF]
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 S. Chabot, L. Salez, F. X. McCormack, L. Touqui, and M. Chignard Surfactant Protein A Inhibits Lipopolysaccharide-Induced In Vivo Production of Interleukin-10 by Mononuclear Phagocytes during Lung Inflammation Am. J. Respir. Cell Mol. Biol., March 1, 2003; 28(3): 347 - 353. [Abstract] [Full Text] [PDF]
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 J. H. Von der Thusen, J. Kuiper, T. J. C. Van Berkel, and E. A. L. Biessen Interleukins in Atherosclerosis: Molecular Pathways and Therapeutic Potential Pharmacol. Rev., March 1, 2003; 55(1): 133 - 166. [Abstract] [Full Text] [PDF]
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 S. Kiriakidis, E. Andreakos, C. Monaco, B. Foxwell, M. Feldmann, and E. Paleolog VEGF expression in human macrophages is NF-{kappa}B-dependent: studies using adenoviruses expressing the endogenous NF-{kappa}B inhibitor I{kappa}B{alpha} and a kinase-defective form of the I{kappa}B kinase 2 J. Cell Sci., February 15, 2003; 116(4): 665 - 674. [Abstract] [Full Text] [PDF]
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P. Sen, S. Bhattacharyya, M. Wallet, C. P. Wong, B. Poligone, M. Sen, A. S. Baldwin Jr., and R. Tisch NF-{kappa}B Hyperactivation Has Differential Effects on the APC Function of Nonobese Diabetic Mouse Macrophages J. Immunol., February 15, 2003; 170(4): 1770 - 1780. [Abstract] [Full Text] [PDF]
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Y. Huang, P. M. Krein, D. A. Muruve, and B. W. Winston Complement Factor B Gene Regulation: Synergistic Effects of TNF-{alpha} and IFN-{gamma} in Macrophages J. Immunol., September 1, 2002; 169(5): 2627 - 2635. [Abstract] [Full Text] [PDF]
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A. Denys, I. A. Udalova, C. Smith, L. M. Williams, C. J. Ciesielski, J. Campbell, C. Andrews, D. Kwaitkowski, and B. M. J. Foxwell Evidence for a Dual Mechanism for IL-10 Suppression of TNF-{alpha} Production That Does Not Involve Inhibition of p38 Mitogen-Activated Protein Kinase or NF-{kappa}B in Primary Human Macrophages J. Immunol., May 15, 2002; 168(10): 4837 - 4845. [Abstract] [Full Text] [PDF]
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 M. CONRON, E. ANDREAKOS, P. PANTELIDIS, C. SMITH, H. L. C. BEYNON, R. M. DUBOIS, and B. M. J. FOXWELL Nuclear Factor-kappa B Activation in Alveolar Macrophages Requires Ikappa B kinase-beta , but Not Nuclear Factor-kappa B Inducing Kinase Am. J. Respir. Crit. Care Med., April 1, 2002; 165(7): 996 - 1004. [Abstract] [Full Text] [PDF]
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W. Lim, W. Ma, K. Gee, S. Aucoin, D. Nandan, F. Diaz-Mitoma, M. Kozlowski, and A. Kumar Distinct Role of p38 and c-Jun N-Terminal Kinases in IL-10-Dependent and IL-10-Independent Regulation of the Costimulatory Molecule B7.2 in Lipopolysaccharide-Stimulated Human Monocytic Cells J. Immunol., February 15, 2002; 168(4): 1759 - 1769. [Abstract] [Full Text] [PDF]
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A. Byrne and D. J. Reen Lipopolysaccharide Induces Rapid Production of IL-10 by Monocytes in the Presence of Apoptotic Neutrophils J. Immunol., February 15, 2002; 168(4): 1968 - 1977. [Abstract] [Full Text] [PDF]
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C. Smith, E. Andreakos, J. B. Crawley, F. M. Brennan, M. Feldmann, and B. M. J. Foxwell NF-{kappa}B-Inducing Kinase Is Dispensable for Activation of NF-{kappa}B in Inflammatory Settings but Essential for Lymphotoxin {beta} Receptor Activation of NF-{kappa}B in Primary Human Fibroblasts J. Immunol., November 15, 2001; 167(10): 5895 - 5903. [Abstract] [Full Text] [PDF]
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 B M J Foxwell, S Yoshimura, J Bondeson, F M Brennan, and M Feldmann High efficiency gene transfer is an efficient way of defining therapeutic targets: a functional genomics approach Ann Rheum Dis, November 1, 2001; 60(90003): iii13 - 17. [Abstract] [Full Text] [PDF]
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J. S. Friedland, D. Constantin, T. C. Shaw, and E. Stylianou Regulation of interleukin-8 gene expression after phagocytosis of zymosan by human monocytic cells J. Leukoc. Biol., September 1, 2001; 70(3): 447 - 454. [Abstract] [Full Text] [PDF]
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A. M. Liese, M. Q. Siddiqi, J. H. Siegel, T. Denny, and Z. Spolarics Augmented TNF-{alpha} and IL-10 production by primed human monocytes following interaction with oxidatively modified autologous erythrocytes J. Leukoc. Biol., August 1, 2001; 70(2): 289 - 296. [Abstract] [Full Text] [PDF]
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M. Conron, J. Bondeson, P. Pantelidis, H. L. C. Beynon, M. Feldmann, R. M. duBois, and B. M. J. Foxwell Alveolar Macrophages and T Cells from Sarcoid, but Not Normal Lung, Are Permissive to Adenovirus Infection and Allow Analysis of NF-kappa B-Dependent Signaling Pathways Am. J. Respir. Cell Mol. Biol., August 1, 2001; 25(2): 141 - 149. [Abstract] [Full Text] [PDF]
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L. Salez, V. Balloy, N. van Rooijen, M. Lebastard, L. Touqui, F. X. McCormack, and M. Chignard Surfactant Protein A Suppresses Lipopolysaccharide-Induced IL-10 Production by Murine Macrophages J. Immunol., May 15, 2001; 166(10): 6376 - 6382. [Abstract] [Full Text] [PDF]
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 R. Rezzonico, V. Imbert, R. Chicheportiche, and J.-M. Dayer Ligation of CD11b and CD11c {beta}2 integrins by antibodies or soluble CD23 induces macrophage inflammatory protein 1{alpha} (MIP-1{alpha}) and MIP-1{beta} production in primary human monocytes through a pathway dependent on nuclear factor-{kappa}B Blood, May 15, 2001; 97(10): 2932 - 2940. [Abstract] [Full Text] [PDF]
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 S. Yoshimura, J. Bondeson, B. M. J. Foxwell, F. M. Brennan, and M. Feldmann Effective antigen presentation by dendritic cells is NF-{{kappa}}B dependent: coordinate regulation of MHC, co-stimulatory molecules and cytokines Int. Immunol., May 1, 2001; 13(5): 675 - 683. [Abstract] [Full Text] [PDF]
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M. D. Wheeler, S. Yamashina, M. Froh, I. Rusyn, and R. G. Thurman Adenoviral gene delivery can inactivate Kupffer cells: role of oxidants in NF-{kappa}B activation and cytokine production J. Leukoc. Biol., April 1, 2001; 69(4): 622 - 630. [Abstract] [Full Text]
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 W. C. Russell Update on adenovirus and its vectors J. Gen. Virol., November 1, 2000; 81(11): 2573 - 2604. [Full Text]
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B. M J Foxwell, J. Bondeson, F. Brennan, and M. Feldmann Adenoviral transgene delivery provides an approach to identifying important molecular processes in inflammation: evidence for heterogenecity in the requirement for NFkappa B in tumour necrosis factor production Ann Rheum Dis, November 1, 2000; 59(90001): i54 - 59. [Abstract] [Full Text] [PDF]
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J. S. Koh, Z. Wang, and J. S. Levine Cytokine Dysregulation Induced by Apoptotic Cells Is a Shared Characteristic of Murine Lupus J. Immunol., October 15, 2000; 165(8): 4190 - 4201. [Abstract] [Full Text] [PDF]
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A. K. De, K. M. Kodys, B. S. Yeh, and C. Miller-Graziano Exaggerated Human Monocyte IL-10 Concomitant to Minimal TNF-{alpha} Induction by Heat-Shock Protein 27 (Hsp27) Suggests Hsp27 Is Primarily an Antiinflammatory Stimulus J. Immunol., October 1, 2000; 165(7): 3951 - 3958. [Abstract] [Full Text] [PDF]
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E. M. Benkhart, M. Siedlar, A. Wedel, T. Werner, and H. W. L. Ziegler-Heitbrock Role of Stat3 in Lipopolysaccharide-Induced IL-10 Gene Expression J. Immunol., August 1, 2000; 165(3): 1612 - 1617. [Abstract] [Full Text] [PDF]
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J. G. Vallejo, P. Knuefermann, D. L. Mann, and N. Sivasubramanian Group B Streptococcus Induces TNF-{alpha} Gene Expression and Activation of the Transcription Factors NF-{kappa}B and Activator Protein-1 in Human Cord Blood Monocytes J. Immunol., July 1, 2000; 165(1): 419 - 425. [Abstract] [Full Text] [PDF]
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H. D. Brightbill, S. E. Plevy, R. L. Modlin, and S. T. Smale A Prominent Role for Sp1 During Lipopolysaccharide- Mediated Induction of the IL-10 Promoter in Macrophages J. Immunol., February 15, 2000; 164(4): 1940 - 1951. [Abstract] [Full Text] [PDF]
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 L. L. Reznikov, B. D. Shames, H. A. Barton, C. H. Selzman, G. Fantuzzi, S.-H. Kim, S. M. Johnson, and C. A. Dinarello Interleukin-1beta deficiency results in reduced NF-kappa B levels in pregnant mice Am J Physiol Regulatory Integrative Comp Physiol, January 1, 2000; 278(1): R263 - R270. [Abstract] [Full Text] [PDF]
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S. Takashiba, T. E. Van Dyke, S. Amar, Y. Murayama, A. W. Soskolne, and L. Shapira Differentiation of Monocytes to Macrophages Primes Cells for Lipopolysaccharide Stimulation via Accumulation of Cytoplasmic Nuclear Factor kappa B Infect. Immun., November 1, 1999; 67(11): 5573 - 5578. [Abstract] [Full Text] [PDF]
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J. Bondeson, B. Foxwell, F. Brennan, and M. Feldmann Defining therapeutic targets by using adenovirus: Blocking NF-kappa B inhibits both inflammatory and destructive mechanisms in rheumatoid synovium but spares anti-inflammatory mediators PNAS, May 11, 1999; 96(10): 5668 - 5673. [Abstract] [Full Text] [PDF]
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W. Ma, W. Lim, K. Gee, S. Aucoin, D. Nandan, M. Kozlowski, F. Diaz-Mitoma, and A. Kumar The p38 Mitogen-activated Kinase Pathway Regulates the Human Interleukin-10 Promoter via the Activation of Sp1 Transcription Factor in Lipopolysaccharide-stimulated Human Macrophages J. Biol. Chem., April 20, 2001; 276(17): 13664 - 13674. [Abstract] [Full Text] [PDF]
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S.-H. Young, J. Ye, D. G. Frazer, X. Shi, and V. Castranova Molecular Mechanism of Tumor Necrosis Factor-alpha Production in 1right-arrow3-beta -Glucan (Zymosan)-activated Macrophages J. Biol. Chem., June 1, 2001; 276(23): 20781 - 20787. [Abstract] [Full Text] [PDF]
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) and zymosan-induced (•) TNF-