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B Inhibitor on In Vitro Cytokine Production and Collagen-Induced Arthritis1
*
Division of Rheumatology, Allergy and Immunology, University of California, San Diego School of Medicine, La Jolla, CA 92093; and
Signal Pharmaceuticals, San Diego, CA 92121
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Abstract |
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 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
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B plays a key role in the production of cytokines in
inflammatory diseases. The effects of a novel T cell-specific NF-B
inhibitor, SP100030, were evaluated in cultured Jurkat cells and in
murine collagen-induced arthritis (CIA). Chemical libraries were
screened for NF-B-inhibitory activity. SP100030, a compound
identified in this process, inhibited NF-B activation in
PMA/PHA-activated Jurkat cells by EMSA at a concentration of 1 µM.
Jurkat cells and the monocytic cell line THP-1 were transfected with an
NF-B promotor/luciferase construct and activated. SP100030 inhibited
luciferase production in the Jurkat cells (IC50 = 30
nM). ELISA and RT-PCR confirmed that IL-2, IL-8, and TNF-
production
by activated Jurkat and other T cell lines were inhibited by SP100030.
However, cytokine expression was not blocked by the compound in THP-1
cells, fibroblasts, endothelial cells, or epithelial cells.
Subsequently, DBA/1J mice were immunized with type II collagen.
Treatment with SP100030 (10 mg/kg/day i.p. beginning on day 21)
significantly decreased arthritis severity from onset of clinical signs
to the end of the study on day 34 (arthritis score, 5.6 ± 1.7 for
SP100030 and 9.8 ± 1.5 for control; p <
0.001). Histologic evaluation demonstrated a trend toward improvement
in SP100030-treated animals. EMSA of arthritic mouse ankles in CIA
showed that synovial NF-B binding was suppressed in the
SP100030-treated mice. SP100030 inhibits NF-B activation in T cells,
resulting in reduced NF-B-regulated gene expression and decreased
CIA. Its selectivity for T cells could provide potent immunosuppression
with less toxicity than other NF-B
inhibitors. |
Introduction |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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, and IL-6, promoting induction of adhesion molecule and
proteinase gene expression (3). These factors play an
important role in attracting and activating other inflammatory cells
and in the degradation of cartilage and bone.
Cytokine and adhesion molecule gene expression is regulated by a
variety of transcription factors. One of these, NF-B, plays an
especially important role in inflammatory processes (4).
The regulation of this particular transcription factor has been
extensively explored in RA in both tissues and cultured cells. For
instance, its expression and activity is markedly increased in the RA
synovium (5, 6). NF-B also plays a pivotal role in IL-6
production by cultured FLS (7). This critical factor is a
potential therapeutic target for the treatment of RA patients.
To assess the possible therapeutic role of NF-B in arthritis, we
investigated a novel T cell-specific transcription factor inhibitor,
SP100030 (8). SP100030 was shown to inhibit NF-B
activation in vitro as well as suppress expression of NF-B-driven
genes. Subsequently, we demonstrated efficacy of this compound in a
chronic arthritis model. These studies suggest that NF-B inhibition
in T cells could be beneficial in inflammatory arthritis.
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Materials and Methods |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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SP100030 was obtained from Signal Pharmaceuticals (San Diego, CA). The chemical formula is 2-chloro-4-(trifluoromethyl)pyrimidine-5-N-(3', 5'-bis(trifluoromethyl)phenyl)-carboxamide. PMA and PHA were obtained from Sigma (St. Louis, MO). A Bandshift kit was purchased from Promega (Madison, WI). Oligonucleotides and Abs cross-reactive to rat p50, p65, c-Rel, RelB, and p52 subunits were purchased from Santa Cruz Biotechnology (Santa Cruz, CA).
Cells
Human Jurkat T cells and THP-monocytes were obtained from the America Type Culture Collection (Manassas, VA) and grown in RPMI 1640 (Media Tech, West Chester, PA) containing 2 mM L-glutamine and 50 U/ml penicillin plus 50 µg/ml streptomycin (Life Technologies, Grand Island, NY) and either 10% FCS (Jurkat) or 20% FCS (Gemini Biological Products, Calabasas, CA) and 2-ME (Sigma) (THP-1). Normal human endothelial cells were obtained from Clonetics (Walkersville, MD), and other cell lines were purchased from American Type Culture Collection (Manassas, VA).
Mice
Six- to 8-wk-old male DBA/1J mice were purchased from The Jackson Laboratory (Bar Harbor, ME). Type II collagen and CFA were obtained from Chondrex (Seattle, WA), IFA and LPS (Escherichia coli serotype 0111:B4) from Sigma.
RT-PCR
Total RNA was isolated using Tri reagent (Molecular Research Center, Cincinnati, OH) according to the manufacturer’s instructions. Semiquantitative RT-PCR was conducted using 2 µg of RNA, 0.4 µM each primer (Clontech, Palo Alto, CA), 0.2 mM each dNTP, 5 µl 10x PCR buffer (100 mM Tris-HCl, 15 mM MgCl2, 500 mM KCl, pH 8.3), 0.5 µl RNasin (40 U/µl; Promega), 0.5 µl avian myeloblastosis virus reverse transcriptase (25 U/µl; Promega), and 0.5 µl Taq polymerase (5 U/µl; Boehringer Mannheim, Chicago, IL) in a 50-µl reaction volume. Reverse transcription was conducted at 42°C for 1 h. The temperature was then adjusted to 95°C for 2 min to inactivate the avian myeloblastosis virus reverse transcriptase. The PCR was amplified for 35 cycles. The PCR cycle consisted of 94°C for 1 min, 55°C for 1 min, and 72°C for 1.5 min. After the final amplification, the reaction was completed at 72°C for 7 min. Twenty microliters of each reaction mix was separated by electrophoresis by using 1.5% agarose gels, followed by staining with ethidium bromide.
NF-B assay
Human Jurkat T cells were stably transfected with a synthetic
promoter comprising three copies of an NF-B binding site (from the
MHC promoter) fused to a minimal SV40 promoter driving luciferase
(9). Cells were counted, resuspended in fresh medium
containing 10% Serum-Plus (HyClone, Logan, UT) at a density of 1
x 106 cells/ml, and plated in 96-well
round-bottom plates (200 µl per well) for 18 h. SP100030 was
dissolved in 0.2% DMSO/H2O and then added at the
appropriate concentrations to the microtiter plates containing the
cells, and the plates were incubated at 37°C for 30 min. To induce
transcriptional activation, 50 ng/ml of PMA and 1 µg/ml of PHA were
added to each well, and the cells were incubated for an additional
5 h at 37°C. Control cells were treated with identical amounts
of DMSO alone. The plates were centrifuged at 2200 rpm for 1 min at
room temperature followed by removal of the media. Sixty microliters of
cell lysis buffer (1x Cell Culture Lysis Reagent, 25 mM
Tris-phosphate, pH 7.8, 2 mM DTT, 2 mM
1,2-diaminocyclohexane-N,N,N',N'-tetraacetic
acid, 10% glycerol, 1% Triton X-100; Promega) was added to each well,
and cells were lysed for 15 min. Then 40 µl of each cell lysate was
transferred to a black 96-well plate, and 50 µl of luciferase
substrate buffer was added. Luminescence was immediately measured using
Packard TopCount (Packard Instruments, Meriden, CT). The ß-actin
assay was run as described above except for that the Jurkat T cells
were transfected with a plasmid that contained the ß-actin promoter
driving luciferase.
Cytokine production assay
After centrifugation, supernatants were collected from each well
in the above luciferase experiments and stored at -20°C until assay.
Approximately 20- to 50-µl aliquots were removed and the levels of
IL-2, IL-6, IL-8, IFN-
, and TNF- were determined by ELISA
(Biosource International, Camarillo, CA).
Collagen-induced arthritis (CIA)
Six- to 8-wk-old male DBA/1J mice were immunized s.c. at the base of the tail with 100 µl of a solution containing type II collagen (1 mg/ml) in CFA and IFA. On day 21, 100 µg of type II collagen in 100 µl PBS was injected i.p. On day 28, 50 µg of LPS in 100 µl PBS was injected i.p. to synchronize and exacerbate arthritis, as previously described (10). Using this protocol, the incidence of arthritis was 100% and peaked at day 34–35. Clinical arthritis scores were evaluated using a scale of 0–4 for each paw (0, normal; 1, minimal erythema and mild swelling; 2, moderate erythema and mild swelling; 3, marked erythema and severe swelling, digits not yet involved; 4, maximal and swelling, digits involved).
Study design
The animals were treated daily starting on day 21 till the end of the study by i.p. injection of either SP100030 (10 mg/kg, dissolved in polyethylene glycol 200 (ICN, Aurora, OH) in a total volume of 50 µl) or 50 µl polyethylene glycol 200 only. The clinical arthritis scores were assessed three times a week in a blinded manner. On day 35, the mice were sacrificed and the two hind paws and one fore paw were harvested from each animal for nuclear protein and RNA extraction, as described below.
Histologic analysis
On day 35, the animals were sacrificed and one forepaw of each animal was fixed in a 10% formalin solution, decalcified, and embedded in paraffin for histologic analysis. Five-micrometer sections were cut, mounted on a glass slide, and stained with hematoxylin and eosin. The tissue was evaluated using a semiquantitative scoring system (0–3+) for synovial hyperplasia, cartilage erosion, and extra-articular inflammation. The maximum score per paw was 9.
Preparation of nuclear extracts of mouse tissue
The mouse paws were cut just above and below the ankle and the skin was removed. The joints were then snap frozen in liquid nitrogen and pulverized. Nuclear extracts were prepared according to the method described by Schreiber et al. with modifications (11). Homogenization of the tissue was performed using 3 ml buffer A (10 mM HEPES, pH 7.9, 1.5 mM MgCl2, 10 mM KCl, 1 mM DTT, 1 mM PMSF, and 0.1% Nonidet P-40). The supernatant was discarded after incubation on ice for 15 min and centrifugation at 850 x g at 4°C. The pellet was resuspended in 4 ml buffer A without Nonidet P-40, the samples were centrifuged, and the supernatant was again discarded. Buffer C (25% (v/v) glycerol, 20 mM HEPES, pH 7.9, 0.42 M NaCl, 1.5 mM MgCl2, 0.2 sodium EDTA, pH 8.0, 1 mM DTT, and 1 mM PMSF; 100 µl) was added to the pellets, and the samples were rocked for 30 min at 4°C. After centrifugation at 4°C, the supernatants were aliquoted and stored at -80°C. The protein concentration was measured using the method of Bradford using a protein dye reagent (12).
EMSA
The Bandshift kit was used according to the manufacturer’s
instructions as previously described (10). Double-stranded
NF-B consensus oligonucleotide probe (5'-AGTTGAGGGGACTTTCCCAGGC-3')
or the mutant oligonucleotide probe (5'-AGTTGAGGCGACTTTCCCAGGC-3')
were end-labeled with [-32P]ATP (50 µCi at
222 TBq/mmol; Amersham Life Science, Arlington Heights, IL). For the
experiments using the Jurkat T cells, binding reactions contained 35
fmol of oligonucleotide and 5 µg of nuclear protein. The reactions
were conducted at room temperature for 20 min in a total volume of 10
µl of binding buffer (10 mM Tris-HCl, pH 7.5, 50 mM NaCl, 1 mM
MgCl2, 0.5 mM EDTA, 0.5 mM DTT, 4% glycerol
(v/v), and 0.5 µg poly(dI-dC) (Pharmacia, Peapack, NJ)). Unlabeled
oligonucleotide was added 5 min before addition of radiolabeled probe
in case of the competition reactions. For supershift analysis, 1 µg
of each Ab was added 5 min before the reaction mixtures immediately
after addition of radiolabeled probe. Following the binding reactions,
1 µl of 10x gel loading buffer was added. For the experiments with
the nuclear extracts of mouse tissue, the samples of the treated and
the untreated groups were pooled. Two microliters of the labeled
oligonucleotides were added to the nuclear extracts (4 µg of protein)
in a total volume of 20 µl, and the samples were incubated 20 min at
room temperature. For the negative control samples, mutant or a 15-fold
excess of cold consensus oligonucleotide was added. To perform
supershift experiments, 2 µg of Abs to NF-B p50 and p65 were
added. Samples were incubated for 15 min at room temperature and loaded
onto a 4% polyacrylamide gel. After electrophoresis, the gel was
transferred to Whatman paper (Whatman International, Maidstone, U.K.)
or vacuum dried and visualized by autoradiography.
Statistical analysis
The clinical and histologic scores were compared using the ANOVA and Student t tests.
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Results |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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B in Jurkat cells is reduced by
SP100030
Chemical libraries were screened to identify compounds that
inhibit induction of NF-B binding activity in stimulated Jurkat
cells. After identification of a lead and subsequent optimization,
SP100030 was identified as a potent NF-B inhibitor and characterized
more completely. To assess the effect of SP100030 on NF-B DNA
binding activity, an EMSA was performed on nuclear extracts of Jurkat
cells that had been stimulated with PMA and PHA for 6 h. As shown
in Fig. 1
, the DNA binding activity was
reduced in the SP100030-treated cells compared with the untreated cells
(lanes 2 and 3, respectively). In
supershift experiments, the DNA protein complex appeared to contain
both p50 and p65. SP100030 decreased the intensity of both the p50 and
p65 supershifted bands. Nonspecific bands with a higher electromobility
were seen in all lanes.
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Having demonstrated that SP100030 decreased NF-B binding
activity, we then determined whether the compound inhibited
transcription of a gene regulated by NF-B. For these experiments,
Jurkat cells were transfected with a construct containing the NF-B
promoter and the luciferase cDNA. Cells were then stimulated with PMA
and PHA to activate NF-B. There was a dose-response-dependent
inhibition of luciferase expression in the transfected Jurkat cells
(see Fig. 2 for a representative
experiment). The IC50 for SP100030 was 30 nM. The
compound had no effect on the ß-actin promoter coupled to luciferase,
demonstrating its specificity for NF-B.
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B driven cytokine gene expression
We subsequently determined whether SP100030 inhibited expression
of NF-B-regulated cytokines in T cells. Jurkat cells were stimulated
with PMA/PHA for 5 h in the presence of SP100030 or vehicle for
0.5–5 h, and cytokine mRNA was determined by RT-PCR analysis. Fig. 3, a representative experiment, shows the
effect of SP100030 on gene expression for IL-2, IL-8, and TNF- in
Jurkat cells. SP100030 markedly decreased IL-2, TNF-, and IL-8 mRNA
levels compared with control cells. Notably, the PCR method used
permits only a semiquantitative assessment of mRNA abundance due to
potential differences in primer efficiency and the relative abundance
of the control gene compared with the cytokine genes. These results
suggest that SP100030 inhibits NF-B-regulated cytokine production at
the transcriptional level.
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The effect of SP100030 on Jurkat cell cytokine protein release was
then determined. PMA/PHA-stimulated Jurkat cells were incubated for
0–5 h in the presence of either SP100030 or vehicle, and the levels of
IL-2, TNF-, and IL-8 were measured in the supernatant by ELISA. Fig. 4A shows that SP100030
inhibited cytokine production in a concentration-dependent fashion in
Jurkat cells. Similar effects were observed in other T cell lines using
several methods to stimulate cells (PMA/PHA; soluble anti-CD3 plus
anti-CD28 Ab; Con A; PMA plus ionophore (A23187)) (see Table I). For each of these lines, complete
dose-responses were evaluated from 0.1 to 30 µM of SP100030. At least
80% inhibition of cytokine production was observed at a
concentration of 3 µM or less (see Table I). In contrast, cytokine
expression was not inhibited by 
10 µM in THP-1 cells (Fig. 4B), other monocytic cell lines, endothelial cells,
fibroblasts, synoviocytes, osteoblasts, and epithelial cell lines (see
Table II). Several different methods of
stimulating these cells was tested, including PMA, LPS, IL-1, or
TNF-, and IL-1 plus TNF-. Although IL-8 was the primary cytokine
evaluated, SP100030 also had no effect on IL-6 and TNF- production
in selected cell lines.
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B activation, EMSA
experiments were performed on U937 and THP-1 cells. SP100030 had no
effect on NF-B binding in the activated monocytic cells.
Furthermore, expression of the NF-B-luciferase construct was not
inhibited by the compound in transfected U937 and U2OS cells (data not
shown). Therapeutic effect of SP100030 in CIA
NF-B activation and subsequent cytokine production are known to
play a role in RA as well as animal models of inflammatory arthritis.
To explore the possibility that T cell-selective NF-B suppression
might have therapeutic efficacy in arthritis, the effect of SP100030
was determined in the murine CIA model. DBA/1J mice were immunized with
type II collagen and treated with either vehicle (PEG 200;
n = 13) or SP100030 (10 mg/kg/day; n =
14) beginning on day 21. Fig. 5 shows
that the SP100030-treated mice had significantly decreased arthritis
scores compared with controls (p < 0.001,
ANOVA). Histologic evaluation of the paws from mice treated from day 20
to 34 showed a trend toward decreased inflammation (mean ± SEM
scores: SP100030, 2.86 ± 1.19; vehicle, 5.73 ± 1.12;
p > 0.05).
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B is inhibited by SP100030
in mice with CIA
We have previously shown that NF-B activity in this model
gradually increases from day 10 to intense NF-B binding at day 35
(10). To determine the effect of SP100030 on NF-B
binding activity in joints, we performed an EMSA on pooled samples from
joints of treated and control mice (Fig. 6). In the group of control animals,
NF-B binding was increased on day 35 (lane 3).
However, the treatment group showed clearly diminished NF-B binding
activity. A portion of the functional NF-B protein consisted of p50,
as determined by supershift experiments, although the supershifted
bands were not as clear as in the human cell line studies.
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Discussion |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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Of the many transcription factors involved in joint inflammation,
NF-B appears to be especially important. For instance, production of
IL-1 and TNF- by synovial macrophages is regulated by NF-B, as is
the expression of TNF- and IL-6 in FLS (15, 16, 17, 18, 19, 20). In
addition, constitutive IL-6 production by RA FLS clones is NF-B
dependent (7). Increased cytokine production driven by
NF-B can enhance expression of vascular adhesion molecules that
attract leukocytes into the joint, as well as matrix
metalloproteinases, which help degrade the extracellular matrix
(21, 22, 23). NF-B protein is readily detected in
rheumatoid synovium, especially in the nuclei of intimal lining cells
(5, 6, 20). Although the proteins are also detected in
osteoarthritis synovium, NF-B DNA binding activity is much greater
in RA than osteoarthritis (10). These data suggest that
this signal transduction pathway can play a pivotal role in the
synovial cytokine cascade.
NF-B is a ubiquitous transcription factor that is primarily
comprised of homo- or heterodimeric combinations of proteins belonging
to the Rel family that shares a highly homologous N-terminal region.
This domain contains sequences required for DNA binding, protein
dimerization, and nuclear localization. Several Rel-containing dimers
exist, of which p50/p65 (NF-B1/RelA) heterodimers and the p50
homodimers are the most common. NF-B normally resides as an inactive
protein in the cytoplasm, where it is bound to an inhibitory protein,
IB. Activation is initiated by many stimuli, including IL-1,
TNF-, platelet-derived growth factor, and phorbol esters. Signal
transduction proceeds through activation of two IB kinases, IKK-1
(IKK-) and IKK-2 (IKK-ß), which phosphorylate IB. These
kinases are constitutively expressed by RA FLS, and IKK-2 stimulation
is necessary and sufficient for NF-B activation in these cells
(20). After phosphorylation and degradation of IB,
NF-B is translocated to the nucleus, where it binds to its target
genes to initiate transcription.
The therapeutic potential for NF-B suppression led us to screen
libraries for compounds that inhibit NF-B activation. After
optimization, one compound, SP100030, was an especially potent
inhibitor of NF-B activation in a T cell line with an
IC50 of 30 nM for NF-B inhibition. Preliminary
pharmacologic studies with SP100030 in rodents demonstrated
antiinflammatory effects in cutaneous delayed-type hypersensitivity and
allogeneic cardiac transplantation (8). The purpose of the
experiments described in the current report was to characterize the in
vitro activity of SP100030 more completely and evaluate its mechanism
of action in a chronic model of inflammatory arthritis in
mice.
After confirming the effect of SP100030 on NF-B activation in Jurkat
cells, we demonstrated that the compound inhibited NF-B-driven
expression of a luciferase reporter gene. This effect could not be
explained by nonspecific toxicity because it had no effect on ß-actin
transcription. Furthermore, expression of endogenous NF-B-regulated
cytokines genes was also suppressed at the mRNA and protein levels by
SP100030. Although this compound blocked NF-B activation in T cell
lines (including Jurkat, Molt-4, HUT-78, and H9), it had no effect on
cytokine production by monocytoid cells, fibroblasts, synoviocytes,
endothelial cells, epithelial cells, and osteoblasts. Additional EMSA
and NF-B-driven reporter gene studies in selected non-T cell lines
confirmed that SP100030 did not suppress NF-B activation. Hence, its
action appears to be relatively specific to T cells. The mechanism of
action is currently under investigation, but does not appear to be
related to IKK inhibition or decreased IB phosphorylation (data
not shown). In any case, the selectivity for the T cell lineage
suggests that SP100030 does not act at signal transduction pathways
that are common to T cells and macrophages.
The effect of SP100030 was then evaluated in murine CIA. We have
previously shown that NF-B is activated in the joints of mice 10
days after systemic immunization with type II collagen even though
clinical arthritis does not appear until day 25–30 (10).
Joint swelling was significantly decreased in animals that were treated
with SP100030 before the onset of joint swelling, although the degree
of disease suppression was surprisingly modest. Clinical efficacy was
accompanied by diminished NF-B activation in joint extracts,
suggesting that the compound acted through this mechanism in vivo.
Although SP100030 only appears to suppress NF-B activation in T
cells, the dependence of synovial inflammation on T cell activation in
CIA likely results in decreased NF-B activation in other cell types
as well. Efficacy in this animal model also indicate that SP100030 can
block NF-B expression in normal cells in addition to the cell lines
tested in vitro. Of interest, animals with established disease did not
appear to benefit when treatment was begun on day 35. At this later
time point, many additional transcription factors, such as AP-1, are
highly activated in CIA and could interfere with efficacy.
Alternatively, T cell-independent processes in established disease
might diminish the effectiveness of a selective T cell approach.
These data suggest that modulation of NF-B activation by compounds
like SP100030 could be beneficial in chronic inflammatory diseases like
arthritis. Of interest, it recently has been shown that the incidence
and severity of CIA was significantly reduced in transgenic mice
expressing a constitutive inhibitor of NF-B/Rel (IB) in the T
cell lineage (24). Selectivity for T cells could also
potentially minimize systemic toxicity that could complicate inhibition
of a key signal transduction pathway. However, in light of the
importance of macrophage-derived cytokines and the relative lack of T
cell products in RA, efficacy in this particular disease would depend
on the ability of SP100030 to suppress local lymphocyte-dependent
responses that support the macrophage-fibroblast cytokine network
(3, 25). Other anti-rheumatic drugs, such as
salicylates, sulfalazine, and corticosteroids also appear to suppress
NF-B activation, possibly contributing to their clinical efficacy
(26, 27, 28). Furthermore, NF-B activation prevents Fas
ligand- and TNF--mediated apoptosis in arthritis and contributes to
synovial hyperplasia (29). Suppression of NF-B in rat
arthritis with NF-B decoy oligonucleotides increased apoptosis in
the inflamed synovium, thereby decreasing swelling. In conjunction with
our studies, these data suggest that NF-B is a potential therapeutic
approach in RA that is amenable to small molecule inhibitors.
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Footnotes |
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2 D.M.G. and L.R. have equally contributed to this study.
3 Current address: Dr. Paul P. Tak, Division of Clinical Immunology and Rheumatology, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands.
4 Address correspondence and reprint requests to Dr. Gary S. Firestein, Division of Rheumatology, Allergy and Immunology No. 0656, University of California at San Diego School of Medicine, 9500 Gilman Drive, La Jolla, CA 92093-0656.
5 Abbreviations used in this paper: RA, rheumatoid arthritis; FLS, fibroblast-like synoviocytes; CIA, collagen-induced arthritis; IKK, IB kinase.
Received for publication January 3, 2000. Accepted for publication May 17, 2000.
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References |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
B: a pivotal transcription factor in chronic inflammatory diseases. N. Engl. J. Med. 336:1066.B in rheumatoid synovium: localization of p50 and p65. Arthritis Rheum. 38:1762.[Medline]
B in human inflamed synovial tissue. Arthritis Rheum. 39:583.[Medline]
B and CBF1. Am. J. Pathol. 152:793.[Abstract]
B- and AP-1-mediated gene expression identified using solution-phase combinatorial chemistry. J. Med. Chem. 41:413.[Medline]
B regulation in rheumatoid arthritis and murine collagen-induced arthritis. Autoimmunity. 28:197.[Medline]
(cA2) versus placebo in rheumatoid arthritis. Lancet 344:1105.[Medline]
B inhibits both inflammatory and destructive mechanisms in rheumatoid synovium but spares anti-inflammatory mediators. Proc. Natl. Acad. Sci. USA 96:5668.B activation in synovial fibroblasts. Ann. Rheum. Dis. 58:55.B regulation by IB kinase in primary fibroblast-like synoviocytes. J. Immunol. 163:427.B/p50 activates an element in the distal matrix metalloproteinase 1 promoter in interleukin-1ß-stimulated synovial fibroblasts. Arthritis Rheum. 41:1987.[Medline]
B activation in collagen-induced arthritis. J. Immunol. 163:1577.B. J. Clin. Invest. 101:1163.[Medline]
B by sodium salicylate and aspirin. Science 265:956.B activation provides the potential link between inflammation and hyperplasia in the arthritic joint. Proc. Natl. Acad. Sci. USA 95:13859.This article has been cited by other articles:
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|
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|
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 U. Eriksson, M. O. Kurrer, I. Sonderegger, G. Iezzi, A. Tafuri, L. Hunziker, S. Suzuki, K. Bachmaier, R. M. Bingisser, J. M. Penninger, et al. Activation of Dendritic Cells through the Interleukin 1 Receptor 1 Is Critical for the Induction of Autoimmune Myocarditis J. Exp. Med., February 3, 2003; 197(3): 323 - 331. [Abstract] [Full Text] [PDF]
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 J. Caamano and C. A. Hunter NF-{kappa}B Family of Transcription Factors: Central Regulators of Innate and Adaptive Immune Functions Clin. Microbiol. Rev., July 1, 2002; 15(3): 414 - 429. [Abstract] [Full Text] [PDF]
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S. G. Harris, R. S. Smith, and R. P. Phipps 15-Deoxy-{Delta}12,1412,14-PGJ2 Induces IL-8 Production in Human T Cells by a Mitogen-Activated Protein Kinase Pathway J. Immunol., February 1, 2002; 168(3): 1372 - 1379. [Abstract] [Full Text] [PDF]
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 L. S. Davis, M. Sackler, R. I. Brezinschek, E. Lightfoot, J. L. Bailey, N. Oppenheimer-Marks, and P. E. Lipsky Inflammation, Immune Reactivity, and Angiogenesis in a Severe Combined Immunodeficiency Model of Rheumatoid Arthritis Am. J. Pathol., January 1, 2002; 160(1): 357 - 367. [Abstract] [Full Text] [PDF]
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 D. C. Jones, X. Ding, and R. A. Daynes Nuclear Receptor Peroxisome Proliferator-activated Receptor alpha (PPARalpha ) Is Expressed in Resting Murine Lymphocytes. THE PPARalpha IN T AND B LYMPHOCYTES IS BOTH TRANSACTIVATION AND TRANSREPRESSION COMPETENT J. Biol. Chem., February 22, 2002; 277(9): 6838 - 6845. [Abstract] [Full Text] [PDF]
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