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HEMOSTASIS, THROMBOSIS, AND VASCULAR BIOLOGY
From the Departments of Immunology and Vascular
Biology, The Scripps Research Institute, La Jolla, CA; the Departments
of Surgery and Physiology and Cellular Physics, College of Physicians
and Surgeons of Columbia University, New York, NY; and the Department
of Neuroanatomy, Institute of Zoology of Jagiellonian University,
Krakow, Poland.
Lipopolysaccharide (LPS) induces human monocytes to express
many proinflammatory mediators, including the procoagulant molecule tissue factor (TF) and the cytokine tumor necrosis factor alpha (TNF- The major outer membrane component of gram-negative
bacteria, lipopolysaccharide (LPS) or endotoxin, is a potent activator of monocyte or macrophage function leading to responses that are both
protective and injurious to the host.1 LPS induces the expression of the procoagulant molecular tissue factor (TF) and inflammatory cytokines such as tumor necrosis factor alpha
(TNF- LPS has been shown to activate members of the mitogen-activated protein
kinase (MAPK) family, including extacellular signal-regulated kinases
(ERK)1/2,6,7 c-Jun amino terminal kinases
(JNKs),8 and p38.9 Inhibition of MAPK kinase
(MEK) in monocytes by a specific inhibitor U0126 reduced LPS induction
of several inflammatory cytokines, including interleukin-1,
interleukin-8, and TNF- LPS stimulation of monocytes activates many transcription factors,
including the nuclear factor NF- The transcription factor Egr-1 is an 80-kd nuclear phosphoprotein
containing 3 zinc-finger DNA-binding domains.16 It is rapidly activated in various cell types in response to a variety of
stimuli.17-19 For instance, LPS stimulation of murine
macrophages induced a rapid induction of Egr-1 transcription, messenger
RNA (mRNA), and protein.20 The Egr-1 promoter contains
several serum response elements (SREs) and Ets binding sites that
mediate induction.21-23 Serum response factor (SRF) and
ternary complex factors (TCFs) form a ternary complex at these SRE and
Ets sites. The TCF proteins are members of the Ets family, which
include Elk-1, Sap1a, and Fli1.23 These proteins are
phosphorylated and activated by upstream MAPKs in a cell type- and
stimulus-specific manner.24,25 Phosphorylation of Elk-1 in
its C-terminal domain induces conformational change that promotes
binding of Elk-1 to Ets sites via its N-terminal Ets domain and binding
to SRF dimers via its B box domain.26 Growth hormone
stimulation of cells leads to phosphorylation of Elk-1 and formation of
an Elk-1/SRF complex that mediates induction of Egr-1 gene
expression.19
We have shown that Egr-1 plays a role in LPS induction of TNF- In this study, we investigated the role of the MEK-ERK1/2 signaling
pathway in LPS induction of TF and TNF- LPS (Escherichia coli serotype 0111:B4) and the MEK
inhibitor PD98059 were obtained from Calbiochem (Carlsbad, CA), and PMA was obtained from Sigma (St Louis, MO).
Cell culture
TNF- TF activity THP-1 or PBMC pellets (1 × 106) were solubilized at 37°C for 15 minutes using 15 mM octyl-D-glucopyranoside. TF activity in cell lysates was assayed using a one-stage clotting assay as described.39 Clotting times were converted to milliunits of TF activity by comparison with a standard curve established with purified human brain TF. For reference, a clotting time of 50 seconds corresponds to 1000 mU of TF activity.Western blotting Cytosolic and nuclear extracts were prepared from THP-1 cells or PBMCs (5 × 106) as described.40 For analysis of TF protein expression, cells were lysed and lysates extracted with 1% Triton X-100 and proteins precipitated with acetone. Proteins were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to Hybond-enhanced chemiluminescence membrane (Amersham Pharmacia Biotech, Alameda, CA). TF was visualized using a 1:1000 dilution of an anti-TF antibody (No. 4501) (American Diagnostica, Greenwich, CT). Egr-1 was visualized using a 1:1000 dilution of an anti-Egr-1 antibody (Santa Cruz Biotechnology, Santa Cruz, CA). Phosphorylated ERK1/2 and total ERK were visualized using a 1:1000 dilution of antibodies against either phosphorylated ERK1/2 or nonphosphorylated ERK1/2, respectively (New England Biolabs, MA). Phosphorylated Elk-1 and nonphosphorylated Elk-1 were visualized using a 1:1000 dilution of antibodies against either phosphorylated Elk-1 (Santa Cruz Biotechnology) or nonphosphorylated Elk-1 (Cell Signaling Technology, Beverly, MA).Northern blotting Total cellular RNA was isolated from THP-1 cells (1 × 107) stimulated with LPS (10 µg/mL) using Trizol Reagent (Gibco). In some experiments, cells were preincubated with PD98059 (50 µM) for 30 minutes at 37°C prior to addition of LPS. RNA (10 µg) was analyzed by Northern blotting as described.41 A 641 human TF complementary DNA (cDNA) fragment, an 800 base pair (bp) human TNF- cDNA fragment, or a 2500 bp human Egr-1 cDNA fragment29 was used as probes. The cDNA
fragments were labeled with [32P], deoxycytidine
triphosphate (ICN, Costa Mesa, CA) using a Prime-It Kit (Stratagene
Cloning Systems, San Diego, CA). Blots were rehybridized with the
housekeeping gene glyceraldehyde 3-phosphate dehydrogenase (G3PDH;
Clontech Laboratories, Palo Alto, CA). Bands were visualized by autoradiography.
Electrophoretic mobility shift assays Nuclear extracts were prepared from THP-1 cells (5 × 106) as described previously.40 Nuclear extracts were incubated with radiolabeled double-stranded oligonucleotide probes (Operon Technologies, Alameda, CA) containing an Egr-1 binding site (underlined), 5'-CCCGGCGCGGGGGCGATTTCGAGTCA-3'; a Sp1 site (underlined), 5'-ATTCGATCGGGGCGGGGCGAGC-3'; the murine immunoglobulin Ig B site (underlined),
5'-CAGAGGGGACTTTCCGAGA-3'; the TFB site (underlined),
5'-GTCCCGGAGTTTCCTACCGGG-3'; an AP-1 site (underlined),
5'-CTGGGGTGAGTCATCCCTT-3'; the Egr-1 SRE3 site (underlined),
5'-AGCACCTTATTTGGAGTGGCCGGATATGGCCCGGCGCTTCC-3'; the Egr-1
SRE4 site (underlined),
5'-AGGATCCCCCGCCGGAACAACCCTTATTTGGGCAG-3'; the Egr-1
SRE5 site (underlined),
5'-TGCGACCCGGAAATGCCATATAAGGAGCAGGAAGGATCCCCT-3'; or
the c-fos SRE site (underlined),
5'-GATCCAGGATGTCCATATTAGGACATCTA-3'. Protein-DNA complexes
were separated from free DNA probe by electrophoresis through 6%
nondenaturing acrylamide gels (Invitrogen, Carlsbad, CA) in 0.5 ×
Tris borate ethylenediaminetetraacetic acid buffer.40 Supershift experiments employed 2 µL each of anti-Egr-1, anti-SRF, anti-Elk-1, and anti-p65 antibodies (Santa Cruz Biotechnology). Recombinant Egr-1 was provided by Dr E. Adamson (Burnham Institute, La
Jolla, CA). Gels were dried, and protein complexes were visualized by autoradiography.
Plasmids The p(B)4-LUC contains 4 copies of an NF-B
site, and p(AP-1)4-LUC contains 4 copies of an AP-1 site.
These sites were cloned upstream of the minimal simian virus 40 (SV40)
promoter expressing the firefly luciferase (LUC) reporter gene in
pGL2-Promoter (Promega, Madison, WI).40,42 The pTF-LUC
contains 2106 bp of the human TF promoter.33 The wild-type
human TF promoter ( 278 to +14) and a version containing mutations in
each of the 3 Egr-1 sites were cloned into pGL2-Basic (Promega,
Madison, WI). The mutations of the Egr-1 sites have been
described.29 The wild-type pTNF--LUC and the Egr-1
mutant contain 615 bp of the human TNF- promoter.27 The
pEgr-1-LUC contains 1200 bp of the murine Egr-1 promoter. The 5'
deletion series of the Egr-1 promoter has been described previously.21 The Egr-1 promoter mutants SRE3 and SRE4
have been described.19 The pEgr-1(SRE5M)LUC
was provided by W. Aird (Harvard University, Boston, MA). The
pcDNA3 (Invitrogen, San Diego, CA) was used as a control
plasmid for transfections. Plasmids expressing dominant-negative
versions of Ras, Raf1, MEK1, ERK1, and ERK2 have been
described.43 In addition, we used plasmid expressing
dominant-negative MEK kinase-1 (MEKK1). The control plasmid pFA-CMV
expresses the GAL4 DNA-binding domain alone and was obtained from
Stratagene Cloning Systems. The pFA2-Elk-1 (pGAL4-Elk-1TA) expresses
the GAL4 DNA-binding domain fused with the transactivation domain of
Elk-1 (Stratagene Cloning Systems). The reporter plasmid pFR-LUC
(pGAL4-LUC) contains 5 copies of the GAL4 binding site upstream of a
minimal promoter that drives expression of the firefly luciferase
reporter gene.
Transfections THP-1 cells were transfected using DEAE-dextran.33 After transfection, cells were incubated in complete media for 46 hours at 37°C before stimulating with LPS (10 µg/mL) for 5 hours at 37°C. In some experiments, cells were preincubated with PD98059 (25 or 50 µM) for 30 minutes at 37°C before the addition of LPS. Cell lysates were assayed for luciferase activity as described in the manufacturer's protocol (Promega) using a Monolight 2010 luminometer (Analytical Luminescence Laboratory, San Diego, CA). In selected experiments, cells were cotransfected with pRLTK, which expresses Renilla luciferase (Promega). Renilla luciferase was measured according to the manufacturer's protocol (Promega) and used to normalize the activity of the firefly luciferase. Variability of the transfection was found to be less than 10%.Data analysis The number of experiments analyzed is indicated in each figure. Band intensities were quantified by densitometric analyses using a Personal Densitometer (Molecular Dynamics, Sunnyvale, CA) and ImageQuant software. Statistical analysis was performed using the unpaired Student t test, and differences were determined to be statistically significant at P < .05.
LPS induction of TF expression requires the MEK-ERK1/2 kinase pathway LPS induces TF and TNF- expression in human monocytes and THP-1
monocytic cells.27,33,44,45 A previous study showed that
activation of the MEK-ERK1/2 pathway was required for LPS induction of
TNF- in human monocytes.6 Therefore, we used TNF- as
a positive control in this study. To determine if the MEK-ERK1/2
pathway is required for LPS induction of TF expression in monocytes, we
measured LPS induction of TF expression in the presence and absence of
the MEK1/2 inhibitor PD98059.46 LPS induction of TF and
TNF- in PBMCs was inhibited in a dose-dependent manner by PD98059
(Figure 1A,B). Similar results were
observed using THP-1 cells (Figures 2A and 3A). Therefore, we used
THP-1 cells as a model of human monocytes to determine how LPS
activation of the MEK-ERK1/2 pathway regulates TF and TNF-
expression. In THP-1 cells, PD98059 inhibited LPS-induced TF protein
expression (Figure 2B) and TF mRNA
expression at both 2 and 4 hours (Figure 2C). The larger TF transcript
represents a partially spliced form that is observed in monocytic
cells.47,48 LPS-induced TNF- mRNA expression was also
inhibited by PD98059 at 2 and 4 hours (Figure
3B). Finally, we examined the effect of
PD98059 on LPS-induced TF and TNF- promoter activity. PD98059 (25 µM) inhibited LPS-induced TF and TNF- promoter activities by 70%
(Figure 2D) and 47% (Figure 3C), respectively. These results indicate
that the MEK-ERK1/2 MAPK pathway plays a major role in the LPS
induction of the TF and TNF- genes in monocytic cells.
Does inhibition of the MEK-ERK1/2 pathway reduce nuclear
activity of NF- B/Rel and AP-1 proteins play a
major role in the LPS induction of TF and TNF- gene expression in
human monocytic cells.27,33,40,49 Therefore, we determined the effect of PD98059 on LPS-stimulated nuclear binding of NF-B and
AP-1 as well as LPS-induced B- and AP-1-dependent transcription in
THP-1 cells. Nuclear translocation was examined by electrophoretic mobility shift assays (EMSAs) using oligonucleotides that contain a
prototypic B site (IgB), a B site from the human TF promoter (TF-B) that binds c-Rel/p65, or an AP-1 site that binds
c-Fos/c-Jun.34 No nuclear NF-/Rel complexes were
detected in unstimulated cells (Figure
4A). LPS induced nuclear translocation,
and binding of both p50/p65 and c-Rel/p65 was not affected by PD98059
(Figure 4A). Next, we determined the effect of PD98059 on LPS-induced B-dependent transcription. THP-1 cells were transiently transfected with a plasmid containing a B-dependent luciferase reporter gene. PD98059 weakly inhibited (29%) LPS-induced, B-dependent
transcription using a reporter plasmid containing a multimerized B
site (Figure 4B). Inhibition of B-dependent transcription by PD98059
may in part contribute to the reduction of LPS-induced TF and TNF-
expression by PD98059. However, the magnitude of this inhibition (29%)
did not appear to account for the stronger inhibition (48%-70%) of PD98059 on LPS-induced TF and TNF- promoter expression.
Cotransfection of the B-dependent reporter plasmid with a plasmid
expressing a dominant-negative ERK1 mutant did not affect LPS-induced,
B-dependent transcription (Figure 4C). Similarly, PD98059 did not
affect the LPS-induced increase in AP-1 binding or AP-1-dependent
transcription (Figure 4D,E). In addition, expression of a
dominant-negative ERK1 mutant did not affect LPS-induced,
AP-1-dependent transcription (Figure 4F). These studies indicate that
inhibition of the MEK-ERK1/2 pathway does not reduce LPS induction of
TF expression by affecting either LPS-induced NF-B or AP-1
activity.
Egr-1 is required for maximal induction of TF and TNF- 111) that contained 3 Egr-1 sites but lacked AP-1 and B sites.33
In addition, DNase I footprinting mapped LPS-inducible footprints to an
Egr-1 site in the TF promoter.35 However, the role of
Egr-1 in TF gene expression in monocytic cells has not been
examined directly.
In this study, we analyzed the kinetics of induction of Egr-1 and TF
mRNA expression in LPS-stimulated THP-1. Maximal levels of Egr-1 mRNA
were observed at 1 hour and preceded the maximal induction of TF mRNA
at 2 hours (Figure 5A). Similarly,
LPS induction of Egr-1 mRNA in mouse peritoneal macrophages
preceded induction of TF mRNA (data not shown). Next, we analyzed the
effect of mutation of Egr-1 sites in the TF and TNF-
LPS induces Egr-1 expression in human monocytes and THP-1 monocytic cells Next, we analyzed if LPS induced Egr-1 expression in human monocytes and THP-1 cells. We found that LPS induced Egr-1 protein in both monocytes and THP-1 cells (Figure 6A,B). Time course experiments using THP-1 cells demonstrated that LPS transiently induced Egr-1 expression with maximal levels in the nucleus at 2 hours (Figure 6C). The newly synthesized Egr-1 protein was functional and bound to an oligonucleotide containing a prototypic Egr-1 sequence (Figure 6D). The Egr-1 complex was competed with an oligonucleotide containing an Egr-1 site and was supershifted with an anti-Egr-1 antibody (data not shown). Sp1 binding was not affected by stimulation with LPS (data not shown). These results confirm that LPS induced Egr-1 expression in both adherent monocytes and THP-1 cells.
LPS induction of Egr-1 expression is mediated by the Ras-Raf-MEK-ERK1/2 pathway To determine if the MEK-ERK1/2 pathway was required for LPS induction of Egr-1, we examined the effect of the MEK-specific inhibitor PD98059 on LPS-induced Egr-1 expression in THP-1 cells. PD98059 inhibited LPS induction of Egr-1 protein and mRNA (Figure 7A,B), and LPS induction of the Egr-1 promoter was also strongly inhibited by PD98059 (Figure 7C). In contrast, another MAPK inhibitor, SB203850, which specifically inhibits p38, had no inhibitory effect on LPS-mediated Egr-1 induction (data not shown). These results indicate that the MEK-ERK1/2 pathway mediates LPS induction of the Egr-1. To complement the experiments using a pharmacologic inhibitor, we determined the effect of dominant-negative mutants of Ras, Raf-1, ERK1, and ERK2 on LPS-induced Egr-1 promoter activity in THP-1 cells. Cotransfection with plasmids expressing dominant-negative versions of Ras, Raf-1, ERK1, or ERK2 reduced LPS induction of Egr-1 promoter activity more than 75% (Figure 7D). In contrast, expression of dominant-negative MEKK-1, an upstream activator of JNK and p38 MAPK, had no significant effect on LPS-induced Egr-1 promoter activity. These results demonstrate the specificity of the Ras-Raf-MEK-ERK1/2 pathway in mediating LPS induction of Egr-1 gene expression in monocytic cells.
Localization of the LPS response element in the Egr-1 promoter The Egr-1 promoter contains 2 distal AP-1 binding sites and 5 SREs (Figure 8A). We used a series of luciferase reporter plasmids containing 5' deletions of the Egr-1 promoter to identify the cis-acting LPS response element in the Egr-1 promoter.21 Deletion of a region containing 2 distal AP-1 sites had no effect on LPS induction of the Egr-1 promoter (Figure 8A). In contrast, deletion of a region containing the distal SRE sites 3 to 5 dramatically decreased Egr-1 promoter activity (Figure 8A). Deletion of a region containing the 2 proximal SRE sites showed no further decrease in promoter activity. These results suggest that SRE sites 3 to 5 mediate LPS induction of the Egr-1 promoter. We therefore examined the effect of individual mutations of the SRE3, SRE4, or SRE5 sites in the Egr-1 promoter on LPS-inducible promoter activity. Mutation of SRE5 showed the strongest inhibition although each of the 3 SRE mutations reduced LPS induction of the Egr-1 promoter (Figure 8B). These results indicate that SRE sites 3 to 5 mediate LPS induction of the Egr-1 promoter in THP-1 cells.
LPS induces formation of an Elk-1/SRF complex at SRE sites SREs are known to bind multicomponent complexes, which contain SRF and TCFs, such as Elk-1, Sap-1, and Fli-1.19,50 Phosphorylation of SRF by various signaling pathways has been shown to increase SRF binding to the SRE.51 TCFs are also targets of MAPK pathways, and phosphorylation-dependent activation of different TCFs has been shown to contribute to Egr-1 gene regulation in cell-type and stimulus-specific manner.52,53 To analyze transcription factors that bind to the SRE sites 3 to 5 in the Egr-1 promoter in monocytic cells, we performed EMSAs with oligonucleotides spanning SRE3, SRE4, and SRE5. An LPS-inducible protein-DNA complex with the same mobility was observed using oligonucleotides spanning each of the SRE sites 3 to 5 (Figure 9A). Additional constitutive protein-DNA complexes were observed with the SRE4 and SRE5.
We performed a detailed characterization of the protein-DNA complexes formed using SRE4. The SRE 4 site bound a protein-DNA complex present in unstimulated THP-1 cells that was competed by an oligonucleotide containing SRE4 but not by an Sp1 site (Figure 9B, upper panel). A similar complex was observed using an oligonucleotide containing the c-fos SRE (data not shown). Sp1 was used as a control and bound to an oligonucleotide containing an Sp1 site. The Sp1 complex was competed with an oligonucleotide containing an Sp1 site but not by an oligonucleotide containing SRE4 (Figure 9B, lower panel). The protein-DNA complexes that bound to SRE4 in unstimulated and LPS-stimulated THP-1 cells could be further separated by performing the electrophoresis at 4°C. The basal complex observed in unstimulated cells could be separated into 2 distinct complexes (Figure 9C, upper panel). The slower migrating basal complex was supershifted with anti-SRF antibody but not by an anti-Elk-1 or an anti-p65 antibody, indicating that it contained SRF but not Elk-1. A faster-migrating nonspecific complex was not recognized by any of the antibodies. Three distinct complexes were observed using nuclear extracts from LPS-stimulated cells that included an inducible complex migrating more slowly than the basal complex. The inducible complex was supershifted with an anti-SRF antibody and an anti-Elk-1 antibody but not by an anti-p65 antibody, indicating that it was a complex composed of SRF and Elk-1 (Figure 9C, lower panel). The LPS-inducible complex observed with SRE5 was also supershifted with antibodies to SRF and Elk-1 (data not shown). LPS also induces activation of JNK1 and phosphorylation of Sap-1.37 However, the LPS-inducible complex was not supershifted with an anti-Sap-1 antibody (data not shown). These results indicate that LPS stimulation of THP-1 cells specifically induced the binding of an Elk-1/SRF complex to the Egr-1 promoter. LPS transiently induces phosphorylation of ERK1/2 and Elk-1 Several studies have shown that LPS rapidly induces ERK1/2 phosphorylation in monocytes and macrophages.6,7,54,55 We examined the time course of ERK1/2 phosphorylation in LPS-stimulated THP-1 monocytic cells. LPS stimulation transiently increased ERK1/2 phosphorylation with a peak at 15 minutes and a return to baseline at 25 minutes (Figure 10A). An antibody against total ERK1/2 was used to demonstrate equal loading of the samples. ERK1/2 has been shown to phosphorylate the transcription factor Elk-1 in fibroblast and HeLa cells.24,25 We therefore determined if LPS stimulation of THP-1 monocytic cells induced phosphorylation of Elk-1. We observed a transient phosphorylation of Elk-1 that peaked at 15 minutes in LPS-stimulated THP-1 cells (Figure 10B). The kinetics of LPS-induced assembly of the Elk-1/SRF complex was also evaluated by EMSAs. LPS transiently increased protein binding to SRE4 with maximal level at 15 minutes (Figure 10C). LPS stimulation did not change Sp1 binding. PD98059 abolished formation of the LPS-induced complex (Figure 10D). These data demonstrated that the kinetics of LPS-induced Elk-1 phosphorylation was identical to the increase in protein binding to SRE4.
LPS activation of the MEK-ERK1/2 pathway mediates phosphorylation of Elk-1 and Elk-1-dependent transcription We finally confirmed if LPS stimulation of THP-1 cells induces ERK1/2-mediated activation of endogenous Elk-1. First, we showed that LPS-induced phosphorylation of Elk-1 was inhibited by PD98059 (Figure 11A). To analyze if LPS induction of the MEK-ERK1/2 pathway can lead to functional activation of Elk-1, THP-1 cells were cotransfected with the reporter plasmid pGAL4-LUC and pGAL4-Elk-1-TA, which expresses a chimeric protein that contains the GAL4 DNA-binding domain fused to the transactivation domain of Elk-1. LPS induced Elk-1-dependent transcription 71-fold, which was strongly inhibited by PD98059 (Figure 11B). No induction was observed using a plasmid that expresses only pGAL4-binding domain (not shown). Next, we determined the effect of dominant-negative versions of Ras, Raf-1, ERK1, and ERK2 on LPS activation of GAL4-Elk-1-TA. Cotransfection with plasmids expressing dominant-negative versions of Ras, Raf-1, ERK1, or ERK2 reduced LPS induction of Elk-1-dependent transcription by 54% to 72% (Figure 11C). In contrast, expression of dominant-negative MEKK-1 had no significant effect on LPS-induced Elk-1-dependent transcription. These results indicate that LPS stimulation of monocytic cells leads to phosphorylation and activation of Elk-1 via the Ras-Raf1-MEK-ERK1/2 pathway.
In this study, we used the MEK1/2 inhibitor PD98059 to show that
LPS activation of the MEK-ERK1/2 pathway was required for the
maximal expression of TF and TNF- The TF and TNF- We demonstrated that LPS rapidly induced Egr-1 expression in human monocytes and THP-1 cells. Inhibition of the Ras-Raf1-MEK-ERK1/2 pathway dramatically reduced LPS induction of Egr-1 promoter activity. Analysis of a 5' deletion series of the Egr-1 promoter localized the cis-acting LPS response element to a region containing SRE sites 3 to 5. Mutation of each of the SRE sites 3 to 5 reduced LPS induction of the Egr-1 promoter, indicating that these sites mediated LPS induction. However, these studies do not exclude the possibility that SRE1 and SRE2 are necessary for the function of these upstream elements. EMSAs demonstrated that SRE3, SRE4, and SRE5 bound an LPS-inducible complex. Supershift experiments indicated that this complex was composed of SRF and Elk-1. We did not observe Sap-1 in the complex. We found that LPS stimulation of cells induced a rapid MEK1/2-dependent phosphorylation of Elk-1 that peaked at 15 minutes and corresponded with the maximal levels of an LPS-inducible protein-DNA complex. Finally, expression of dominant-negative mutants of the Ras-ERK1/2 pathway and the use of PD98059 demonstrated that LPS stimulation of monocytic cells induced Elk-1-dependent transcription via the Ras-Raf1-MEK-ERK1/2 pathway. Dominant-negative MEKK1 did not reduce LPS induction of either the Egr-1 promoter or the Elk-1-dependent reporter gene, suggesting that the JNK MAPK pathway does not play a significant role in LPS induction of Egr-1 in monocytic cells. A model of LPS induction of TF in monocytic cells is shown in Figure
12. The current study demonstrated that
LPS stimulation of monocytic cells leads to the rapid MEK1/2-dependent
phosphorylation of Elk-1. An LPS-inducible complex consisting of Elk-1
and SRF mediated induction of Egr-1 gene expression. We propose that
newly synthesized Egr-1 binds to the TF promoter and contributes to maximal induction of TF expression in LPS-stimulated cells. LPS stimulation of monocytic cells also activated the JNK MAPK
pathway. The JNK pathway phosphorylates and activates AP-1
proteins bound to the TF promoter.56 Finally, LPS
stimulation activates IKK
We thank R. Clarkson and W. Aird for mutated versions of the Egr-1 promoter, and J. Robertson and B. Parker for preparing the manuscript.
Submitted February 5, 2001; accepted April 25, 2001.
Supported by grants from the National Institutes of Health (HL48872) to N.M. and a postdoctoral fellowship from the American Heart Association (M.A.O.).
M.G. and M.A.O. contributed equally to the study.
The publication costs of this article were defrayed in part by page charge payment. Therefore, and solely to indicate this fact, this article is hereby marked "advertisement" in accordance with 18 U.S.C. section 1734.
Presented in abstract form at the 42nd annual meeting of the American Society of Hematology, December 1-5, 2000, San Francisco, CA. Reprints: Nigel Mackman, Dept of Immunology, C-204, The Scripps Research Institute, 10550 North Torrey Pines Rd, La Jolla, CA 92037; e-mail: nmackman{at}scripps.edu.
1. Ulevitch RJ, Tobias PS. Recognition of gram-negative bacteria and endotoxin by the innate immune system. Curr Opin Immunol. 1999;11:19-22[CrossRef][Medline] [Order article via Infotrieve]. 2. Müller JM, Ziegler-Heitbrock HWL, Baeuerle PA. Nuclear factor kappa B, a mediator of lipopolysaccharide effects. Immunobiology. 1993;187:233-256[Medline] [Order article via Infotrieve].
3.
Steinemann S, Ulevitch RJ, Mackman N.
Role of the lipopolysaccharide (LPS)-binding protein/CD14 pathway in LPS induction of tissue factor expression in monocytic cells.
Arterioscler Thromb.
1994;14:1202-1209
4.
Poltorak A, He X, Smirnova I, et al.
Defective LPS signaling in C3H/HeJ and C57BL/10ScCr mice: mutation in Tlr4 gene.
Science.
1998;282:2085-2088 5. Beutler B. Tlr4: central component of the sole mammalian LPS sensor. Curr Opin Immunol. 2000;12:20-26[CrossRef][Medline] [Order article via Infotrieve].
6.
van der Bruggen T, Nijenhuis S, van Raaij E, Verhoef J, van Asbeck BS.
Lipopolysaccharide-induced tumor necrosis factor alpha production by human monocytes involves the Raf-1/MEK1-MEK2/ERK1-ERk2 pathway.
Infect Immun.
1999;67:3824-3829 7. Liu MK, Herrera-Velit P, Brownsey RW, Reiner NE. CD-14-dependent activation of protein kinase C and mitogen-activated protein kinases (p42 and p44) in human monocytes treated with bacterial lipopolysaccharide. J Immunol. 1994;153:2642-2652[Abstract].
8.
Hambleton J, Weinstein SL, Lem L, DeFranco AL.
Activation of c-Jun N-terminal kinase in bacterial lipopolysaccharide-stimulated macrophages.
Proc Natl Acad Sci U S A.
1996;93:2774-2778
9.
Casey J, Petranka JG, Kottra J, Fleenor DE, Rosse WF.
The structure of the urokinase-type plasminogen activator receptor gene.
Blood.
1994;84:1151-1156
10.
Scherle PA, Jones EA, Favata MF, et al.
Inhibition of MAP kinase kinase prevents cytokine and prostaglandin E2 production in lipopolysaccharide-stimulated monocytes.
J Immunol.
1998;161:5681-5686
11.
Baeuerle PA, Baltimore D.
NF-
12.
May MJ, Ghosh S.
I
13.
O'Connell MA, Bennett BL, Mercurio F, Manning AM, Mackman N.
Role of IKK1 and IKK2 in lipopolysaccharide signaling in human monocytic cells.
J Biol Chem.
1998;273:30410-30414
14.
Swantek JL, Christerson L, Cobb MH.
Lipopolysacchride-induced tumor necrosis factor-
15.
Fischer C, Page S, Weber M, Eisele T, Neumeier D, Brand K.
Differential effects of lipopolysaccharide and tumor necrosis factor on monocytic I 16. Sukhatme VP. Early transcriptional events in cell growth: the Egr family. J Am Soc Nephrol. 1990;1:859-866[Abstract]. 17. Schwachtgen J-L, Houston P, Campbell C, Sukhatme V, Braddock M. Fluid shear stress activation of egr-1 transcription in cultured human endothelial and epithelial cells is mediated via the extracellular signal-related kinase 1/2 mitogen-activated protein kinase pathway. J Clin Invest. 1998;101:2540-2549[Medline] [Order article via Infotrieve].
18.
Santiago FS, Lowe HC, Day FL, Chesterman CN, Khachigian LM.
Early growth response factor-1 induction by injury is triggered by release and paracrine activation by fibroblast growth factor-2.
Am J Pathol.
1999;154:937-944
19.
Clarkson RW, Shang CA, Levitt LK, Howard T, Waters MJ.
Ternary complex factors Elk-1 and Sap-1a mediate growth hormone-induced transcription of egr-1 (early growth response factor-1) in 3T3-F442A preadipocytes.
Mol Endocrinol.
1999;13:619-631 20. Coleman DL, Bartiss AH, Sukhatme VP, Liu J, Rupprecht HD. Lipopolysaccharide induces Egr-1 mRNA and protein in murine peritoneal macrophages. J Immunol. 1992;149:3045-3051[Abstract].
21.
Cohen DM, Gullans SR, Chin WW.
Urea inducibility of egr-1 in murine inner medullary collecting duct cells is mediated by the serum response element and adjacent Ets motifs.
J Biol Chem.
1996;271:12903-12908 22. McMahon SB, Monroe JG. A ternary complex factor-dependent mechanism mediates induction of egr-1 through selective serum response elements following antigen receptor cross-linking in B lymphocytes. Mol Cell Biol. 1995;15:1086-1093[Abstract]. 23. Watson DK, Robinson L, Hodge DR, Kola I, Papas TS, Seth A. FLI1 and EWS-FLI1 function as ternary complex factors and ELK1 and SAP1a function as ternary and quaternary complex factors on the Egr1 promoter serum response elements. Oncogene. 1997;14:213-221[CrossRef][Medline] [Order article via Infotrieve]. 24. Janknecht R, Ernst WH, Pingoud V, Nordheim A. Activation of ternary complex factor Elk-1 by MAP kinases. EMBO J. 1993;12:5097-5104[Medline] [Order article via Infotrieve]. 25. Sharrocks AD. ERK2/p42 MAP kinase stimulates both autonomous and SRF-dependent DNA binding by Elk-1. FEBS Lett. 1995;368:77-80[CrossRef][Medline] [Order article via Infotrieve]. 26. Yang SH, Shore P, Willingham N, Lakey JH, Sharrocks AD. The mechanism of phosphorylation-inducible activation of the ETS-domain transcription factor Elk-1. EMBO J. 1999;18:5666-5674[CrossRef][Medline] [Order article via Infotrieve].
27.
Yao J, Mackman N, Edgington TS, Fan S-T.
Lipopolysaccharide induction of the tumor necrosis factor-
28.
Mechtcheriakova D, Wlachos A, Holzmuller H, Binder BR, Hofer E.
Vascular endothelial cell growth factor-induced tissue factor expression in endothelial cells is mediated by EGR-1.
Blood.
1999;93:3811-3823
29.
Cui MZ, Parry GC, Oeth P, et al.
Transcriptional regulation of the tissue factor gene in human epithelial cells is mediated by Sp1 and EGR-1.
J Biol Chem.
1996;271:2731-2739
30.
Houston P, Dickson MC, Ludbrook V, et al.
Fluid shear stress induction of the tissue factor promoter in vitro and in vivo is mediated by Egr-1.
Arterioscler Thromb Vasc Biol.
1999;19:281-289
31.
Yan SF, Zou YS, Gao Y, et al.
Tissue factor transcription driven by Egr-1 is a critical mechanism of murine pulmonary fibrin deposition in hypoxia.
Proc Natl Acad Sci U S A.
1998;95:8298-8303
32.
Cui MZ, Penn MS, Chisolm GM.
Native and oxidized low density lipoprotein induction of tissue factor gene expression in smooth muscle cells is mediated by both Egr-1 and Sp1.
J Biol Chem.
1999;274:32795-32802
33.
Mackman N, Brand K, Edgington TS.
Lipopolysaccharide-mediated transcriptional activation of the human tissue factor gene in THP-1 monocytic cells requires both activator protein 1 and nuclear factor
34.
Oeth P, Parry GCN, Mackman N.
Regulation of the tissue factor gene in human monocytic cells: role of AP-1, NF-
35.
Groupp ER, Donovan-Peluso M.
Lipopolysaccharide induction of THP-1 cells activates binding of c-Jun, Ets, and Egr-1 to the tissue factor promoter.
J Biol Chem.
1996;271:12423-12430 36. Cruzalegui FH, Cano E, Treisman R. ERK activation induces phosphorylation of Elk-1 at multiple S/T-P motifs to high stoichiometry. Oncogene. 1999;18:7948-7957[CrossRef][Medline] [Order article via Infotrieve]. 37. Janknecht R, Hunter T. Convergence of MAP kinase pathways on the ternary complex factor Sap-1a. EMBO J. 1997;16:1620-1627[CrossRef][Medline] [Order article via Infotrieve]. 38. Boyum A. Isolation of mononuclear cells and granulocytes from human blood: isolation of mononuclear cells by one centrifugation, and of granulocytes by combining centrifugation and sedimentation at 1 g. Scand J Clin Lab Invest Suppl. 1968;97:77-89[Medline] [Order article via Infotrieve]. 39. Morrissey JH, Fair DS, Edgington TS. Monoclonal antibody analysis of purified and cell-associated tissue factor. Thromb Res. 1988;52:247-261[CrossRef][Medline] [Order article via Infotrieve].
40.
Oeth PA, Parry GCN, Kunsch C, Nantermet P, Rosen CA, Mackman N.
Lipopolysaccharide induction of tissue factor gene expression in monocytic cells is mediated by binding of c-Rel/p65 heterodimers to a
41.
Kirkland TN, Virca GD, Kuus-Reichel T, et al.
Identification of lipopolysaccharide binding proteins in 70Z/3 cells by photoaffinity cross-linking.
J Biol Chem.
1990;265:9520-9525
42.
Oeth P, Yao J, Fan S-T, Mackman N.
Retinoic acid selectively inhibits lipopolysaccharide induction of tissue factor gene expression in human monocytes.
Blood.
1998;91:2857-2865
43.
Yan SF, Lu J, Zou YS, et al.
Hypoxia-associated induction of early growth response-1 gene expression.
J Biol Chem.
1999;274:15030-15040 44. Burchett SK, Weaver WM, Westall JA, Larsen A, Kronheim S, Wilson CB. Regulation of tumor necrosis factor/cachectin and IL-1 secretion in human mononuclear phagocytes. J Immunol. 1988;140:3473-3481[Abstract].
45.
Gregory SA, Morrissey JH, Edgington TS.
Regulation of tissue factor gene expression in the monocyte procoagulant response to endotoxin.
Mol Cell Biol.
1989;9:2752-2755
46.
Dudley DT, Pang L, Decker SJ, Bridges AJ, Saltiel AR.
A synthetic inhibitor of the mitogen-activated protein kinase cascade.
Proc Natl Acad Sci U S A.
1995;92:7686-7689
47.
Brand K, Fowler BJ, Edgington TS, Mackman N.
Tissue factor mRNA in THP-1 monocytic cells is regulated at both transcriptional and posttranscriptional levels in response to lipopolysaccharide.
Mol Cell Biol.
1991;11:4732-4738 48. van der Logt CPE, Reitsma PH, Bertina RM. Alternative splicing is responsible for the presence of two tissue factor mRNA species in LPS stimulated human monocytes. Thromb Haemost. 1992;67:272-276[Medline] [Order article via Infotrieve].
49.
Trede NS, Tsytsykova AV, Chatila T, Goldfeld AE, Geha RS.
Transcriptional activation of the human TNF- 50. Marais R, Wynne J, Treisman R. The SRF accessory protein Elk-1 contains a growth factor-regulated transcriptional activation domain. Cell. 1993;73:381-393[CrossRef][Medline] [Order article via Infotrieve]. 51. Janknecht R, Hipskind RA, Houthaeve T, Nordheim A, Stunnenberg HG. Identification of multiple SRF N-terminal phosphorylation sites affecting DNA binding properties. EMBO J. 1992;11:1045-1054[Medline] [Order article via Infotrieve].
52.
Mora-Garcia P, Sakamoto KM.
Granulocyte colony-stimulating factor induces egr-1 up-regulation through interaction of serum response element-binding proteins.
J Biol Chem.
2000;275:22418-22426 53. McMahon SB, Monroe JG. The role of early growth response gene 1 (egr-1) in regulation of the immune response. J Leukoc Biol. 1996;60:159-166[Abstract].
54.
Marie C, Roman-Roman S, Rawadi G.
Involvement of mitogen-activated protein kinase pathways in interleukin-8 production by human monocytes and polymorphonuclear cells stimulated with lipopolysaccharide or Mycoplasma fermentans membrane lipoproteins.
Infect Immun.
1999;67:688-693 55. Geppert TD, Whitehurst CE, Thompson P, Beutler B. Lipopolysaccharide signals activation of tumor necrosis factor biosynthesis through the ras/raf-1/MEK/MAPK pathway. Mol Med. 1994;1:93-103[Medline] [Order article via Infotrieve].
56.
Hall AJ, Vos HL, Bertina RM.
Lipopolysaccharide induction of tissue factor in THP-1 cells involves Jun protein phosphorylation and nuclear factor
57.
Cogswell PC, Mayo MW, Baldwin AS Jr.
Involvement of Egr-1/RelA synergy in distinguishing T cell activation from tumor necrosis factor-
© 2001 by The American Society of Hematology.
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  S. Y. Shin, H. Song, C. G. Kim, Y.-K. Choi, K. S. Lee, S.-J. Lee, H.-J. Lee, Y. Lim, and Y. H. Lee Egr-1 Is Necessary for Fibroblast Growth Factor-2-induced Transcriptional Activation of the Glial Cell Line-derived Neurotrophic Factor in Murine Astrocytes J. Biol. Chem., October 30, 2009; 284(44): 30583 - 30593. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P.-H. Park, H. Huang, M. R. McMullen, P. Mandal, L. Sun, and L. E. Nagy Suppression of Lipopolysaccharide-stimulated Tumor Necrosis Factor-{alpha} Production by Adiponectin Is Mediated by Transcriptional and Post-transcriptional Mechanisms J. Biol. Chem., October 3, 2008; 283(40): 26850 - 26858. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 B. K. Pedersen and M. A. Febbraio Muscle as an Endocrine Organ: Focus on Muscle-Derived Interleukin-6 Physiol Rev, October 1, 2008; 88(4): 1379 - 1406. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. Hoffmann, J. Ashouri, S. Wolter, A. Doerrie, O. Dittrich-Breiholz, H. Schneider, E. F. Wagner, J. Troppmair, N. Mackman, and M. Kracht Transcriptional Regulation of EGR-1 by the Interleukin-1-JNK-MKK7-c-Jun Pathway J. Biol. Chem., May 2, 2008; 283(18): 12120 - 12128. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S.-J. Jiang, C.-C. Kuo, M. W. Berry, A. W. Lee, and L. A. Campbell Identification and Characterization of Chlamydia pneumoniae-Specific Proteins That Activate Tumor Necrosis Factor Alpha Production in RAW 264.7 Murine Macrophages Infect. Immun., April 1, 2008; 76(4): 1558 - 1564. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. P. Luyendyk, G. A. Schabbauer, M. Tencati, T. Holscher, R. Pawlinski, and N. Mackman Genetic Analysis of the Role of the PI3K-Akt Pathway in Lipopolysaccharide-Induced Cytokine and Tissue Factor Gene Expression in Monocytes/Macrophages J. Immunol., March 15, 2008; 180(6): 4218 - 4226. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 B. H. Choi, C. G. Kim, Y.-S. Bae, Y. Lim, Y. H. Lee, and S. Y. Shin p21Waf1/Cip1 Expression by Curcumin in U-87MG Human Glioma Cells: Role of Early Growth Response-1 Expression Cancer Res., March 1, 2008; 68(5): 1369 - 1377. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. S. Andrew, V. Bernardo, L. A. Warnke, J. C. Davey, T. Hampton, R. A. Mason, J. E. Thorpe, M. A. Ihnat, and J. W. Hamilton Exposure to Arsenic at Levels Found in U.S. Drinking Water Modifies Expression in the Mouse Lung Toxicol. Sci., November 1, 2007; 100(1): 75 - 87. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Miksa, R. Wu, X. Cui, W. Dong, P. Das, H. H. Simms, T. S. Ravikumar, and P. Wang Vasoactive Hormone Adrenomedullin and Its Binding Protein: Anti-Inflammatory Effects by Up-Regulating Peroxisome Proliferator-Activated Receptor-{gamma} J. Immunol., November 1, 2007; 179(9): 6263 - 6272. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 L. I. Pahlman, E. Malmstrom, M. Morgelin, and H. Herwald M protein from Streptococcus pyogenes induces tissue factor expression and pro-coagulant activity in human monocytes Microbiology, August 1, 2007; 153(8): 2458 - 2464. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 P. Goldin-Lang, K. Pels, Q.-V. Tran, B. Szotowski, F. Wittchen, S. Antoniak, T. Willich, H. Witt, M. Hummel, D. Lenze, et al. Effect of ionizing radiation on cellular procoagulability and co-ordinated gene alterations Haematologica, August 1, 2007; 92(8): 1091 - 1098. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. P. Luyendyk, J. D. Piper, M. Tencati, K. V. Reddy, T. Holscher, R. Zhang, J. Luchoomun, X. Chen, W. Min, C. Kunsch, et al. A Novel Class of Antioxidants Inhibit LPS Induction of Tissue Factor by Selective Inhibition of the Activation of ASK1 and MAP Kinases Arterioscler Thromb Vasc Biol, August 1, 2007; 27(8): 1857 - 1863. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 W.-F. Fang, J. H. Cho, Q. He, M.-C. Lin, C.-C. Wu, N. F. Voelkel, and I. S. Douglas Lipid A fraction of LPS induces a discrete MAPK activation in acute lung injury Am J Physiol Lung Cell Mol Physiol, August 1, 2007; 293(2): L336 - L344. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P.-h. Park, M. R. McMullen, H. Huang, V. Thakur, and L. E. Nagy Short-term Treatment of RAW264.7 Macrophages with Adiponectin Increases Tumor Necrosis Factor-{alpha} (TNF-{alpha}) Expression via ERK1/2 Activation and Egr-1 Expression: ROLE OF TNF-{alpha} IN ADIPONECTIN-STIMULATED INTERLEUKIN-10 PRODUCTION J. Biol. Chem., July 27, 2007; 282(30): 21695 - 21703. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. V. Gafencu, M. R. Robciuc, E. Fuior, V. I. Zannis, D. Kardassis, and M. Simionescu Inflammatory Signaling Pathways Regulating ApoE Gene Expression in Macrophages J. Biol. Chem., July 27, 2007; 282(30): 21776 - 21785. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. Ahamed, F. Niessen, T. Kurokawa, Y. K. Lee, G. Bhattacharjee, J. H. Morrissey, and W. Ruf Regulation of macrophage procoagulant responses by the tissue factor cytoplasmic domain in endotoxemia Blood, June 15, 2007; 109(12): 5251 - 5259. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. V. Tsytsykova, J. V. Falvo, M. Schmidt-Supprian, G. Courtois, D. Thanos, and A. E. Goldfeld Post-induction, Stimulus-specific Regulation of Tumor Necrosis Factor mRNA Expression J. Biol. Chem., April 20, 2007; 282(16): 11629 - 11638. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Cai, C. Song, I. Endoh, J. Goyette, W. Jessup, S. B. Freedman, H. P. McNeil, and C. L. Geczy Serum Amyloid A Induces Monocyte Tissue Factor J. Immunol., February 1, 2007; 178(3): 1852 - 1860. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. L. Sherry, J. C. O'Connor, J. M. Kramer, and G. G. Freund Augmented Lipopolysaccharide-Induced TNF-{alpha} Production by Peritoneal Macrophages in Type 2 Diabetic Mice Is Dependent on Elevated Glucose and Requires p38 MAPK J. Immunol., January 15, 2007; 178(2): 663 - 670. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J.-H. Kang, M.-J. Kim, H.-I. Jang, K.-H. Koh, K.-S. Yum, D.-J. Rhie, S. H. Yoon, S. J. Hahn, M.-S. Kim, and Y.-H. Jo Proximal cyclic AMP response element is essential for exendin-4 induction of rat EGR-1 gene Am J Physiol Endocrinol Metab, January 1, 2007; 292(1): E215 - E222. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Ke, M. Gururajan, A. Kumar, A. Simmons, L. Turcios, R. L. Chelvarajan, D. M. Cohen, D. L. Wiest, J. G. Monroe, and S. Bondada The Role of MAPKs in B Cell Receptor-induced Down-regulation of Egr-1 in Immature B Lymphoma Cells J. Biol. Chem., December 29, 2006; 281(52): 39806 - 39818. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. M. Egorina, M. A. Sovershaev, T. V. Kondratiev, J. O. Olsen, T. Tveita, and B. Osterud Induction of Monocytic Tissue Factor Expression After Rewarming From Hypothermia In Vivo Is Counteracted by Heat Shock in c-Jun-Dependent Manner Arterioscler Thromb Vasc Biol, October 1, 2006; 26(10): 2401 - 2406. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 N. McNamara, M. Gallup, A. Sucher, I. Maltseva, D. McKemy, and C. Basbaum AsialoGM1 and TLR5 Cooperate in Flagellin-Induced Nucleotide Signaling to Activate Erk1/2 Am. J. Respir. Cell Mol. Biol., June 1, 2006; 34(6): 653 - 660. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 V. Thakur, M. T. Pritchard, M. R. McMullen, Q. Wang, and L. E. Nagy Chronic ethanol feeding increases activation of NADPH oxidase by lipopolysaccharide in rat Kupffer cells: role of increased reactive oxygen in LPS-stimulated ERK1/2 activation and TNF-{alpha} production J. Leukoc. Biol., June 1, 2006; 79(6): 1348 - 1356. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 L. Servillo, C. Balestrieri, A. Giovane, P. Pari, D. Palma, G. Giannattasio, M. Triggiani, and M. L. Balestrieri Lysophospholipid Transacetylase in the Regulation of Paf Levels in Human Monocytes and Macrophages FASEB J, May 1, 2006; 20(7): 1015 - 1017. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. Li, M. R. Power, and T.-J. Lin De novo synthesis of early growth response factor-1 is required for the full responsiveness of mast cells to produce TNF and IL-13 by IgE and antigen stimulation Blood, April 1, 2006; 107(7): 2814 - 2820. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Mishra, T. Fujita, V. N. Lama, D. Nam, H. Liao, M. Okada, K. Minamoto, Y. Yoshikawa, H. Harada, and D. J. Pinsky Carbon monoxide rescues ischemic lungs by interrupting MAPK-driven expression of early growth response 1 gene and its downstream target genes PNAS, March 28, 2006; 103(13): 5191 - 5196. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Banerjee, R. Gugasyan, M. McMahon, and S. Gerondakis Diverse Toll-like receptors utilize Tpl2 to activate extracellular signal-regulated kinase (ERK) in hemopoietic cells PNAS, February 28, 2006; 103(9): 3274 - 3279. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Chanchevalap, M. O. Nandan, B. B. McConnell, L. Charrier, D. Merlin, J. P. Katz, and V. W. Yang Kruppel-like factor 5 is an important mediator for lipopolysaccharide-induced proinflammatory response in intestinal epithelial cells Nucleic Acids Res., February 25, 2006; 34(4): 1216 - 1223. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 L. M. Khachigian Early Growth Response-1 in Cardiovascular Pathobiology Circ. Res., February 3, 2006; 98(2): 186 - 191. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T. Vowinkel, K. C. Wood, K. Y. Stokes, J. Russell, C. F. Krieglstein, and D. N. Granger Differential expression and regulation of murine CD40 in regional vascular beds Am J Physiol Heart Circ Physiol, February 1, 2006; 290(2): H631 - H639. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J.-G. Wang, S. A. Mahmud, J. A. Thompson, J.-G. Geng, N. S. Key, and A. Slungaard The principal eosinophil peroxidase product, HOSCN, is a uniquely potent phagocyte oxidant inducer of endothelial cell tissue factor activity: a potential mechanism for thrombosis in eosinophilic inflammatory states Blood, January 15, 2006; 107(2): 558 - 565. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Eligini, F. Violi, C. Banfi, S. S. Barbieri, M. Brambilla, M. Saliola, E. Tremoli, and S. Colli Indobufen inhibits tissue factor in human monocytes through a thromboxane-mediated mechanism Cardiovasc Res, January 1, 2006; 69(1): 218 - 226. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Boelen, J. Kwakkel, A. Alkemade, R. Renckens, E. Kaptein, G. Kuiper, W. M. Wiersinga, and T. J. Visser Induction of Type 3 Deiodinase Activity in Inflammatory Cells of Mice with Chronic Local Inflammation Endocrinology, December 1, 2005; 146(12): 5128 - 5134. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. A. Rushworth, X.-L. Chen, N. Mackman, R. M. Ogborne, and M. A. O'Connell Lipopolysaccharide-Induced Heme Oxygenase-1 Expression in Human Monocytic Cells Is Mediated via Nrf2 and Protein Kinase C J. Immunol., October 1, 2005; 175(7): 4408 - 4415. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. M. Ogborne, S. A. Rushworth, and M. A. O'Connell {alpha}-Lipoic Acid-Induced Heme Oxygenase-1 Expression Is Mediated by Nuclear Factor Erythroid 2-Related Factor 2 and p38 Mitogen-Activated Protein Kinase in Human Monocytic Cells Arterioscler Thromb Vasc Biol, October 1, 2005; 25(10): 2100 - 2105. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Kamimura, C. Viedt, A. Dalpke, M. E. Rosenfeld, N. Mackman, D. M. Cohen, E. Blessing, M. Preusch, C. M. Weber, J. Kreuzer, et al. Interleukin-10 Suppresses Tissue Factor Expression in Lipopolysaccharide-Stimulated Macrophages via Inhibition of Egr-1 and a Serum Response Element/MEK-ERK1/2 Pathway Circ. Res., August 19, 2005; 97(4): 305 - 313. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Li-Weber, M. K. Treiber, M. Giaisi, K. Palfi, N. Stephan, S. Parg, and P. H. Krammer Ultraviolet Irradiation Suppresses T Cell Activation via Blocking TCR-Mediated ERK and NF-{kappa}B Signaling Pathways J. Immunol., August 15, 2005; 175(4): 2132 - 2143. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. Strassheim, J.-Y. Kim, J.-S. Park, S. Mitra, and E. Abraham Involvement of SHIP in TLR2-Induced Neutrophil Activation and Acute Lung Injury J. Immunol., June 15, 2005; 174(12): 8064 - 8071. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. Molor-Erdene, K. Okajima, H. Isobe, M. Uchiba, N. Harada, and H. Okabe Urinary trypsin inhibitor reduces LPS-induced hypotension by suppressing tumor necrosis factor-{alpha} production through inhibition of Egr-1 expression Am J Physiol Heart Circ Physiol, March 1, 2005; 288(3): H1265 - H1271. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 H. Shinohara, A. Inoue, N. Toyama-Sorimachi, Y. Nagai, T. Yasuda, H. Suzuki, R. Horai, Y. Iwakura, T. Yamamoto, H. Karasuyama, et al. Dok-1 and Dok-2 are negative regulators of lipopolysaccharide-induced signaling J. Exp. Med., February 7, 2005; 201(3): 333 - 339. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Mostecki, B. M. Showalter, and P. B. Rothman Early Growth Response-1 Regulates Lipopolysaccharide-induced Suppressor of Cytokine Signaling-1 Transcription J. Biol. Chem., January 28, 2005; 280(4): 2596 - 2605. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 V. Korcheva, J. Wong, C. Corless, M. Iordanov, and B. Magun Administration of Ricin Induces a Severe Inflammatory Response via Nonredundant Stimulation of ERK, JNK, and P38 MAPK and Provides a Mouse Model of Hemolytic Uremic Syndrome Am. J. Pathol., January 1, 2005; 166(1): 323 - 339. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 W. Wu, R. D. Mosteller, and D. Broek Sphingosine Kinase Protects Lipopolysaccharide-Activated Macrophages from Apoptosis Mol. Cell. Biol., September 1, 2004; 24(17): 7359 - 7369. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
X. Wu, G. A. Zimmerman, S. M. Prescott, and D. M. Stafforini The p38 MAPK Pathway Mediates Transcriptional Activation of the Plasma Platelet-activating Factor Acetylhydrolase Gene in Macrophages Stimulated with Lipopolysaccharide J. Biol. Chem., August 20, 2004; 279(34): 36158 - 36165. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Epelman, D. Stack, C. Bell, E. Wong, G. G. Neely, S. Krutzik, K. Miyake, P. Kubes, L. D. Zbytnuik, L. L. Ma, et al. Different Domains of Pseudomonas aeruginosa Exoenzyme S Activate Distinct TLRs J. Immunol., August 1, 2004; 173(3): 2031 - 2040. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. V. Reddy, G. Bhattacharjee, G. Schabbauer, A. Hollis, K. Kempf, M. Tencati, M. O'Connell, M. Guha, and N. Mackman Dexamethasone enhances LPS induction of tissue factor expression in human monocytic cells by increasing tissue factor mRNA stability J. Leukoc. Biol., July 1, 2004; 76(1): 145 - 151. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. H. Rhee, A. C. Keates, M. P. Moyer, and C. Pothoulakis MEK Is a Key Modulator for TLR5-induced Interleukin-8 and MIP3{alpha} Gene Expression in Non-transformed Human Colonic Epithelial Cells J. Biol. Chem., June 11, 2004; 279(24): 25179 - 25188. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Jiang, C. Van de Ven, P. Satwani, L. V. Baxi, and M. S. Cairo Differential Gene Expression Patterns by Oligonucleotide Microarray of Basal versus Lipopolysaccharide-Activated Monocytes from Cord Blood versus Adult Peripheral Blood J. Immunol., May 15, 2004; 172(10): 5870 - 5879. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. R. Yan, G. Qing, D. M. Byers, A. W. Stadnyk, W. Al-Hertani, and R. Bortolussi Role of MyD88 in Diminished Tumor Necrosis Factor Alpha Production by Newborn Mononuclear Cells in Response to Lipopolysaccharide Infect. Immun., March 1, 2004; 72(3): 1223 - 1229. [Abstract] [Full Text] [PDF]
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T. Minami, A. Sugiyama, S.-Q. Wu, R. Abid, T. Kodama, and W. C. Aird Thrombin and Phenotypic Modulation of the Endothelium Arterioscler Thromb Vasc Biol, January 1, 2004; 24(1): 41 - 53. [Abstract] [Full Text] [PDF]
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F. Bea, E. Blessing, B. J Bennett, C. C. Kuo, L. A. Campbell, J. Kreuzer, and M. E Rosenfeld Chronic inhibition of cyclooxygenase-2 does not alter plaque composition in a mouse model of advanced unstable atherosclerosis Cardiovasc Res, October 15, 2003; 60(1): 198 - 204. [Abstract] [Full Text] [PDF]
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 L. E. Nagy Recent Insights into the Role of the Innate Immune System in the Development of Alcoholic Liver Disease Experimental Biology and Medicine, September 1, 2003; 228(8): 882 - 890. [Abstract] [Full Text] [PDF]
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H. Takeya, E. C. Gabazza, S. Aoki, H. Ueno, and K. Suzuki Synergistic effect of sphingosine 1-phosphate on thrombin-induced tissue factor expression in endothelial cells Blood, September 1, 2003; 102(5): 1693 - 1700. [Abstract] [Full Text] [PDF]
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 H.-P. Tzeng, F.-M. Ho, K.-F. Chao, M.-L. Kuo, S.-Y. Lin-Shiau, and S.-H. Liu {beta}-Lapachone Reduces Endotoxin-induced Macrophage Activation and Lung Edema and Mortality Am. J. Respir. Crit. Care Med., July 1, 2003; 168(1): 85 - 91. [Abstract] [Full Text] [PDF]
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 B. Ji, X.-q. Chen, D. E. Misek, R. Kuick, S. Hanash, S. Ernst, R. Najarian, and C. D. Logsdon Pancreatic gene expression during the initiation of acute pancreatitis: identification of EGR-1 as a key regulator Physiol Genomics, June 24, 2003; 14(1): 59 - 72. [Abstract] [Full Text] [PDF]
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J. Rupp and M. Maass Egr-1, a Major Link Between Infection and Atherosclerosis? Circ. Res., May 16, 2003; 92 (9): e78 - e78. [Full Text] [PDF]
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R. K. Giri, S. K. Selvaraj, and V. K. Kalra Amyloid Peptide-Induced Cytokine and Chemokine Expression in THP-1 Monocytes Is Blocked by Small Inhibitory RNA Duplexes for Early Growth Response-1 Messenger RNA J. Immunol., May 15, 2003; 170(10): 5281 - 5294. [Abstract] [Full Text] [PDF]
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R. Pawlinski, B. Pedersen, B. Kehrle, W. C. Aird, R. D. Frank, M. Guha, and N. Mackman Regulation of tissue factor and inflammatory mediators by Egr-1 in a mouse endotoxemia model Blood, May 15, 2003; 101(10): 3940 - 3947. [Abstract] [Full Text] [PDF]
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 D. L. Russell, K. M. H. Doyle, I. Gonzales-Robayna, C. Pipaon, and J. S. Richards Egr-1 Induction in Rat Granulosa Cells by Follicle-Stimulating Hormone and Luteinizing Hormone: Combinatorial Regulation By Transcription Factors Cyclic Adenosine 3',5'-Monophosphate Regulatory Element Binding Protein, Serum Response Factor, Sp1, and Early Growth Response Factor-1 Mol. Endocrinol., April 1, 2003; 17(4): 520 - 533. [Abstract] [Full Text] [PDF]
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 T. Morichika, H. K. Takahashi, H. Iwagaki, T. Yoshino, R. Tamura, M. Yokoyama, S. Mori, T. Akagi, M. Nishibori, and N. Tanaka Histamine Inhibits Lipopolysaccharide-Induced Tumor Necrosis Factor-{alpha} Production in an Intercellular Adhesion Molecule-1- and B7.1-Dependent Manner J. Pharmacol. Exp. Ther., February 1, 2003; 304(2): 624 - 633. [Abstract] [Full Text] [PDF]
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S.-Q. Wu, T. Minami, D. J. Donovan, and W. C. Aird The proximal serum response element in the Egr-1 promoter mediates response to thrombin in primary human endothelial cells Blood, December 15, 2002; 100(13): 4454 - 4461. [Abstract] [Full Text] [PDF]
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Y. Zen, K. Harada, M. Sasaki, K. Tsuneyama, K. Katayanagi, Y. Yamamoto, and Y. Nakanuma Lipopolysaccharide Induces Overexpression of MUC2 and MUC5AC in Cultured Biliary Epithelial Cells : Possible Key Phenomenon of Hepatolithiasis Am. J. Pathol., October 1, 2002; 161(4): 1475 - 1484. [Abstract] [Full Text] [PDF]
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M. Guha and N. Mackman The Phosphatidylinositol 3-Kinase-Akt Pathway Limits Lipopolysaccharide Activation of Signaling Pathways and Expression of Inflammatory Mediators in Human Monocytic Cells J. Biol. Chem., August 23, 2002; 277(35): 32124 - 32132. [Abstract] [Full Text] [PDF]
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 K.-E. Eilertsen and B. Osterud The central role of thromboxane and platelet activating factor receptors in ex vivo regulation of endotoxin-induced monocyte tissue factor activity in human whole blood Innate Immunity, August 1, 2002; 8(4): 285 - 293. [Abstract] [PDF]
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U. Bavendiek, P. Libby, M. Kilbride, R. Reynolds, N. Mackman, and U. Schonbeck Induction of Tissue Factor Expression in Human Endothelial Cells by CD40 Ligand Is Mediated via Activator Protein 1, Nuclear Factor kappa B, and Egr-1 J. Biol. Chem., July 5, 2002; 277(28): 25032 - 25039. [Abstract] [Full Text] [PDF]
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L. Shi, R. Kishore, M. R. McMullen, and L. E. Nagy Chronic Ethanol Increases Lipopolysaccharide-stimulated Egr-1 Expression in RAW 264.7 Macrophages. CONTRIBUTION TO ENHANCED TUMOR NECROSIS FACTOR alpha PRODUCTION J. Biol. Chem., April 19, 2002; 277(17): 14777 - 14785. [Abstract] [Full Text] [PDF]
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H. Kato Regulation of Functions of Vascular Wall Cells by Tissue Factor Pathway Inhibitor: Basic and Clinical Aspects Arterioscler Thromb Vasc Biol, April 1, 2002; 22(4): 539 - 548. [Abstract] [Full Text] [PDF]
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 L. Shi, R. Kishore, M. R. McMullen, and L. E. Nagy Lipopolysaccharide stimulation of ERK1/2 increases TNF-alpha production via Egr-1 Am J Physiol Cell Physiol, June 1, 2002; 282(6): C1205 - C1211. [Abstract] [Full Text] [PDF]
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H. Kato Regulation of Functions of Vascular Wall Cells by Tissue Factor Pathway Inhibitor: Basic and Clinical Aspects Arterioscler Thromb Vasc Biol, April 1, 2002; 22(4): 539 - 548. [Abstract] [Full Text] [PDF]
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| Copyright © 2001 by American Society of Hematology Online ISSN: 1528-0020 | |||||||||
expression by inducing Elk-1 phosphorylation and Egr-1
expression
Top
Abstract
Introduction
is required for 
