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B Site in the Proximal Promoter
Reproductive Endocrinology Division, Department of Obstetrics and Gynecology, University of Michigan (D.I.L.), Ann Arbor, Michigan 48109-0276; Center for Reproductive Sciences, Department of Obstetrics, Gynecology, and Reproductive Sciences, University of California (V.A.C., J.-F.M., R.N.T.), San Francisco, California 94143-0556
Address all correspondence and requests for reprints to: Robert N. Taylor, M.D., Ph.D., Center for Reproductive Sciences, Department of Obstetrics, Gynecology, and Reproductive Sciences, University of California, HSE 1689, Box 0556, San Francisco, California 94143-0556. E-mail: rtaylor{at}socrates.ucsf.edu
Abstract
A complex network of cytokines mediates immunoregulatory responses
in the pathogenesis of endometriosis. RANTES (regulated upon
activation, normal T cell expressed and secreted) is a chemoattractant
for monocytes and T cells. Endometriotic lesions express RANTES, and
its concentration in peritoneal fluid correlates with the severity of
endometriosis. We investigated the influence of IL-1ß, a potent
macrophage cytokine, on RANTES production in endometriotic stromal
cells and determined the region of the RANTES promoter responsible for
IL-1ß action. RANTES mRNA was induced 5-fold in endometriotic stromal
cells, and the conditioned medium RANTES protein concentrations were
12-fold higher in IL-1ß-treated endometriotic stromal cells
vs. untreated controls (P < 0.05).
IL-1ß activated the full-length (-940 bp) RANTES promoter as well as
a truncated 456-bp 5'-flanking construct by 2-fold. Mutagenesis of a
nuclear factor-
B response element at -30 bp abolished the IL-1ß
effect, whereas mutation of a nearby TNF response element did not
affect the IL-1ß induction. An IL-1ß time-course Western assay
revealed a rapid diminution of IB (endogenous inhibitor of nuclear
factor-B) in endometriotic stromal cells. Overexpression of IB in
endometriotic stromal cells inhibited the IL-1ß response of the
RANTES gene promoter. Transcription of RANTES mRNA is up-regulated by
IL-1ß via a nuclear factor-B response element in the proximal
RANTES gene promoter. These results demonstrate a feed-forward
regulatory loop in the pathogenesis of endometriosis by which IL-1ß
produced from activated macrophages can lead to further macrophage
recruitment via RANTES production in endometriotic stromal
cells.
ONE CATALYST IN the pathogenesis of endometriosis is an enhanced immune response surrounding the ectopic implants. Activated macrophages are the predominant leukocytes (1, 2), and their secretory products are increased in the peritoneal fluid of endometriosis-affected women (3, 4). Therefore, we chose to study a principal macrophage-derived cytokine, IL-1ß, and its role in propagating endometriotic implants through a proinflammatory stimulus, synthesis of the CC-chemokine RANTES (regulated on activation, normal T-cell expressed and secreted).
RANTES, originally isolated as a cDNA from CD8+ T
cells (5), is an 8-kDa secreted protein with
chemoattractant actions on monocytes, NK cells, T cells, and
eosinophils (6, 7). Several cell types, including
fibroblasts, epithelial cells, and macrophages themselves, express
RANTES within hours of stimulation by proinflammatory stimuli, such as
TNF
and interferon-
(8). We reported the
production of RANTES in stromal cells of endometriotic lesions
(9) and observed a positive correlation between the
severity of endometriosis and the concentration of RANTES in peritoneal
fluid (10).
IL-1ß is proposed to play a central role in the integrated inflammatory cascade associated with endometriosis (11). In addition, we have shown that IL-1ß activates an angiogenic phenotype in endometriotic stromal cells by inducing the production of proangiogenic proteins, vascular endothelial growth factor and IL-6 (12). As IL-1ß was reported to induce RANTES production in other cell types (13, 14, 15), we investigated whether IL-1ß could induce RANTES expression in isolated primary endometriotic stromal cells (ESC).
Activation of the transcription factor nuclear factor (NF)-B by
proinflammatory stimuli leads to increased expression of several genes
involved in inflammation. In the present study we show that IL-1ß
induced RANTES gene expression and protein secretion in ESC. Evaluation
of the human RANTES gene promoter revealed four potential NF-B
response elements. One of these, lying just upstream of the
transcription start site, was identified as essential for the IL-1ß
response in ESC.
Materials and Methods
Sources of tissues
Tissue specimens were obtained from patients undergoing
laparoscopy or laparotomy after providing written informed consent
under a study protocol approved by the University of California-San
Francisco Committee on Human Research. Healthy ovulatory women, who had
not received hormones or GnRH agonist therapy for at least 6 months
before surgery, were recruited. Women with endometriosis (mean ±
SD age, 35 ± 5 yr; n = 4) were staged
intraoperatively according to the revised American Society for
Reproductive Medicine classification (16). All women were
classified as stage III, based on the presence of an endometrioma 3 cm
or greater in diameter. Endometrioma biopsies were collected under
sterile conditions and transported to the laboratory on ice in MEM
with 10% FBS. All samples were examined histologically and considered
endometriotic lesions when epithelium and stroma were seen.
Human endometriotic cell cultures
Primary endometriotic cell cultures were prepared from endometrioma biopsies, as we have described previously (17). Glandular epithelial cells were separated from stromal cells and debris by filtration through narrow gauge sieves. ESC were subcultured to eliminate contamination by macrophages or other leukocytes. Extensive characterization of the ESC cultures prepared using this protocol confirmed that they were more than 95% pure and retained functional markers of their endometrial origin in vitro (17).
IL-1ß stimulation
Cultures of ESC were plated in 10-cm culture dishes
(Becton Dickinson and Co., Lincoln Park, NJ) and allowed
to grow to confluence in 10% FBS-supplemented medium. Before the
addition of cytokine, the medium was changed to a low serum medium
(MEM supplemented with 2.5% FBS, antibiotics, nucleosides, and
nonessential amino acids). Pilot dose-response experiments showed a
maximum stimulation after treatment with recombinant human IL-1ß (10
ng/ml = 0.6 nM; Sigma, St. Louis, MO).
Conditioned media were removed and analyzed after 4, 8, 12, and 24
h. Pilot experiments showed that 85% of maximal RANTES accumulation
was reached after 12 h. The 2.5% FBS-supplemented MEM used for
the experiments was tested for IL-1ß and RANTES concentrations, and
both were below the limit of detection for the respective ELISA.
Preparation of total RNA and Northern analysis
Total RNA was extracted from cell cultures using the TRIzol reagent kit (Life Technologies, Inc., Gaithersburg, MD). Total RNA (10 µg) was subjected to electrophoresis on agarose gels and blotted by capillary transfer onto nylon membranes (Schleicher & Schuell, Inc., Keene, NH). The membranes were hybridized with a 32P-labeled RANTES complementary DNA (cDNA) probe synthesized by random primer extension (CLONTECH Laboratories, Inc., Palo Alto, CA). The template for the RANTES probe is a 165-bp fragment of RANTES cDNA (18). The integrity and relative amount of RNA loaded into each lane were confirmed using a 240-bp 32P-labeled glyceraldehyde-3-phosphate dehydrogenase (GAPDH) cDNA probe as a constitutively expressed marker. Data were analyzed as ratios of the density of the hybridization signals of RANTES to GAPDH mRNA, as determined by a PhosphorImager (Storm, Molecular Dynamics, Inc., Menlo Park, CA).
RANTES ELISA
A specific sandwich ELISA was used to quantify RANTES in conditioned media (Quantikine, R&D Systems, Minneapolis, MN). In our laboratory the assay was linear and sensitive to 1 pM, with intra- and interassay coefficients of variation of 3.2% and 6.5%, respectively. The assay is specific for human RANTES, with no known cross-reactivity with other cytokines or chemokines (R&D Systems, 2000 Immunoassay Catalog). Aliquots of culture supernatants were each tested in duplicate at several dilutions and compared with reference standards of recombinant human RANTES (R&D Systems).
Reporter genes and expression vectors
The full-length RANTES promoter (19) was subcloned into the pGL2 vector (Promega Corp., Madison, WI). Mutants were generated with QuickChange site-directed mutagenesis kits (Stratagene), using oligonucleotides containing the desired mutation. All constructs and mutants were sequenced by the University of California-San Francisco Biomolecular Resource Center to verify that the correct sequences or mutations were present.
Cell transfections and luciferase assays
Transient transfections for luciferase assays were conducted in
human ESC grown in MEM with 10% FBS and antibiotics. Subconfluent
cells were collected by centrifugation, and the pellet was resuspended
with Dulbecco’s PBS (0.5 ml/1.5 x 107
cells) containing 0.1% dextrose, 10 µg/ml Biobrene, and respective
reporter plasmids. Ten micrograms of pGL2-RANTES promoter (firefly
luciferase, experimental reporter) and the suspended cells were
transferred to a cuvette and kept at room temperature for 5 min. The
cells were electroporated using a Bio-Rad Laboratories, Inc., gene pulser set at 300 V and 975 microfarads. The
electroporated cells were then transferred to 6.5 ml MEM with 10%
FBS and antibiotics. The cells were recovered in the medium for 5 min,
and then resuspended and plated at 0.25 ml/dish in 24-well multiplates.
IL-1ß was added to octuplicate wells to a final concentration of 0.6
nM in a total volume of 0.50 ml/dish. The cells were then
incubated for 18 h. Using this method the transfection efficiency
obtained with ß-galactosidase reporter vectors in ESC was 40%.
Cell extracts were prepared with reporter lysis buffer (Promega Corp.) after washing, and then luciferase activity were measured with a commercial kit from Promega Corp. Each reporter vector was assayed in at least three independent cultures. The luciferase transfection efficiencies were normalized to an independent control plasmid (2.0 µg Renilla luciferase reporter). The results are presented as the fold increase in luciferase activity (±SEM) between untreated cells (controls) and cells treated with 0.6 nM IL-1ß.
IB inactivation
Cells were plated in 12-well plates at 150,000 cells/ml in 10% FBS and antibiotics. Medium was replaced the next day, and 24 h later subconfluent cultures were stimulated with 0.6 nM IL-1ß for the indicated times. Incubations were terminated by aspiration of the medium, two washes with ice-cold PBS, and addition of 200 µl lysis buffer [1% Triton X-100 lysis buffer containing 20 mM Tris-HCl (pH 8.0), 137 mM NaCl, 10% (vol/vol) glycerol, 2 mM EDTA, and 1 mM Pefabloc (0.14 U aprotinin, 20 µM leupeptin, and 1 mM sodium orthovanadate)] at 4 C.
To detect the transient inactivation of IB, equal quantities of cell
lysates (15 µg) were resolved by SDS-PAGE (12%) and transferred to
polyvinylidene difluoride membranes (Millipore Corp.,
Bedford, MA). Western blots were performed using a rabbit polyclonal
antihuman IB antibody (C-terminal peptide, 1:2500 dilution, gift
from Dr. W. Greene, San Francisco General Hospital, San Francisco,
CA) that recognizes the 38-kDa form of IB. Western blots were
incubated with the antibody and then washed in Tris-buffered saline
containing 0.1% Tween 20. Antigen-antibody complexes were detected
with horseradish peroxidase-coupled secondary antibodies and the
enhanced chemiluminescence system (Renaissance, NEN Life Science Products, Boston, MA). Finally, the blot was developed on
reflection NEF film (NEN Life Science Products).
Statistical analysis
All experiments were repeated a minimum of three times and analyzed by ANOVA, unpaired or paired t tests as appropriate. Results are presented as the mean ± SEM. Significant differences were accepted when two-tailed analyses yielded P < 0.05 (20). NS indicates no significant difference from control conditions.
Results
RANTES mRNA expression in ESC
Four independent ESC preparations were evaluated for the
expression of RANTES mRNA after IL-1ß stimulation. Northern
hybridization was used to identify and quantify RANTES mRNA transcripts
(Fig. 1
). The RANTES probe detected a
single transcript of 1.3 kb, in agreement with other reports
(5). This probe also showed low level cross-hybridization
with 28S and 18S rRNA bands on the total RNA blots, but these could be
easily distinguished from the RANTES mRNA signal. As an internal
control for RNA quantity and integrity, the blots were reprobed to
quantify mRNA representing the constitutive GAPDH gene (1.2 kb), which
was used to normalize the phosphorimaging data. Relative to untreated
ESC, incubation for 12 h in the presence of IL-1ß (0.6
nM) resulted in a 5.0-fold increase in steady state
RANTES mRNA levels in ESC.
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Enhanced secretion of immunoreactive RANTES protein from ESC
exposed to IL-1ß (0.6 nM) was confirmed by ELISA. The
ratios of IL-1ß-treated to untreated cells showed a 12-fold increase
in RANTES secretion after cytokine stimulation (34 ± 8
vs. 3 ± 1 pM; P
< 0.05; Fig. 2).
|
Gene promoter-reporter constructs were transiently transfected
into ESC to test the hypothesis that IL-1ß regulated RANTES
expression at the transcriptional level. Transfection of the parent
vector (pGL2) alone had low basal activity and showed no IL-1ß
response (Fig. 3). When the full-length
RANTES promoter (940 bp of 5'-flanking DNA relative to the
transcriptional start site) or a truncated version (456 bp of
5'-flanking DNA) was used for transfection, IL-1ß treatment resulted
in a 2-fold increase in transgene activation (2.2 ± 0.3 and
2.1 ± 0.2, respectively; P < 0.05).
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To assess the role of specific domains within the RANTES promoter,
ESCs were transiently transfected with wild-type RANTES promoter
constructs (456 bp of 5'-flanking DNA) or mutated promoter sequences
linked to the luciferase reporter gene. The specific mutations are
described in Fig. 4. In cells transfected
with the wild-type promoter, treatment with IL-1ß led to a 2.1
± 0.2-fold stimulation of transcription (Fig. 5). When a single TNF response element
(TNFre) site (21) present in the RANTES promoter sequence
(-164) was altered by site-directed mutagenesis (see Fig. 4), the
2-fold IL-1ß effect persisted (2.0 ± 0.4; P <
0.05). However, when the proximal NF-B response element (-40) of
the RANTES promoter was mutated in a similar fashion, the IL-1ß
effect was abolished (1.1 ± 0.1; NS relative to control; Fig. 5).
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B Western analysis after IL-1ß treatment of ESC
NF-B exists in the cytoplasm in an inactive form associated
with antagonistic regulatory proteins called inhibitors of B
(IB). The activation of NF-B is associated with phosphorylation
of IB, followed by its degradation and release from NF-B and
nuclear translocation of NF-B. To verify the activation of the
NF-B pathway by IL-1ß in ESC, cells were treated with the cytokine
over a 1-h time course and subjected to Western analysis (Fig. 6). Specific antibodies against IB
demonstrated a rapid (t1/2, <5 min), transient
depletion of ESC cytoplasmic IB. Levels of the protein begin to
return toward baseline by 45–60 min. The kinetics of degradation are
comparable to that induced by IL-1ß in other cell lines
(22). This finding is consistent with a rapid
NF-B-mediated induction of RANTES gene transcription after
IL-1ß stimulation.
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B prevents IL-1ß induction of RANTES gene
transcription
To confirm that the NF-B pathway was necessary for IL-1ß
induction of RANTES gene transcription, we cotransfected ESC with
wild-type RANTES promoter plasmids and an expression vector driving the
overproduction of IB. In the presence of high intracellular IB
levels, IL-1ß failed to activate the RANTES gene promoter (1.2
± 0.1, NS relative to control; Fig. 7).
IB expression had no effect on the basal activation of the RANTES
reporter, thus demonstrating no intrinsic inhibitory activity (Fig. 7).
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Endometriotic lesions are accompanied by an inflammatory response within the peritoneal cavity and the implants themselves (11). IL-1ß is present in elevated concentrations in peritoneal fluid of patients with endometriosis compared with those without endometriosis and may play a pathophysiological role in the disease (23, 24, 25). The objective of the current study was to evaluate the effect of IL-1ß on RANTES gene expression in ESC.
RANTES is a chemotactic cytokine, or chemokine, secreted from a variety
of cell types, including ESC. Its role in the host defense system is to
recruit specific leukocytes, particularly macrophages, to sites of
ongoing inflammation and injury (26). We (10)
and others (27, 28) initially postulated that monocyte
chemoattraction might explain the predominance of this cell type in the
peritoneal fluid of subjects with endometriosis. Immunoreactive
(10) and bioactive RANTES concentrations (29)
in peritoneal fluid correlate positively with the clinical stage
of endometriosis. Furthermore, RANTES protein is expressed in
endometriotic lesions in situ (9). As other
cell types express RANTES in response to IL-1ß (13, 15, 30, 31), including normal human endometrial stromal cells
(32), we chose to look at the regulation of RANTES
expression by IL-1ß in stromal cells derived from endometriotic
lesions. Previously we showed that ESC are more responsive to IL-1ß
stimulation than cells derived from normal endometrium
(12). We hypothesized that the regulation of RANTES in ESC
was mediated by the activation of inducible transcription factors
binding to specific promoter sequences. The findings of our current
study indicate that IL-1ß stimulates the production and secretion of
RANTES in ESC via activation of the proximal NF-B response element
in the RANTES gene promoter.
In other cell types RANTES expression is regulated at the level of gene
transcription and to a lesser degree by posttranscriptional mechanisms
(8, 33, 34, 35). This may explain the discrepancy between
IL-1ß-induced mRNA levels (5-fold) and apparent transcriptional
activation of the RANTES promoter (2-fold) observed in this report. The
inducible transcription factor NF-B is essential for the expression
of many genes involved in inflammatory responses (36). The
Rel/NF-B proteins (p65, Rel-B, c-Rel, p50, and p52) comprise a
family of structurally related inducible transcription factors that
bind DNA as dimers and whose activity is regulated by subcellular
location (37). NF-B in the cytoplasm is complexed to an
inhibitor protein called IB (37). Activation signals
initiate protein kinase cascades that result in the phosphorylation of
the IB-kinase complex, which, on degradation by the proteasome,
permits NF-B to translocate into the nucleus and bind its
recognition elements within the promoter regions of NF-B-responsive
genes (38). Previous studies in human uterine cells
demonstrated IB degradation and NF-B nuclear translocation with
kinetics identical to those reported in our paper (39).
Accordingly, we propose that IL-1ß influences the site-specific
recruitment of leukocytes through activation of an NF-B response
element on the RANTES gene promoter in ESC.
The human RANTES promoter region contains two NF-B binding sites at
positions -54 and -40 relative to the transcription start site
(40). These two elements are typical NF-B binding sites
that bind the p65/p50 heterodimer. In ESC, IL-1ß induced the RANTES
promoter via an NF-B cis-acting binding site located
within the -40 to -31 region of the RANTES promoter.
Using activated peripheral blood lymphocytes, Nelson et al.
(41) abolished the RANTES promoter activity by mutating a
putative NF-B site located upstream from the region mutated in our
study. Génin et al. (42) transfected U937
cells with RANTES promoter constructs and found a
trans-activation requirement for both the same NF-B site
we identified as well as an upstream interferon--stimulated response
element. Reduced IL-1ß-induced RANTES gene activity was found in an
astrocytoma cell line when point mutations were introduced at a
location similar to ours, although additional factor(s) binding from
-278 to -194 also mediated the IL-1ß effect in these cells
(31). Roebuck et al. (43)
described how similar proinflammatory stimuli can differentially
regulate CC and CXC chemokine gene promoters. Thus, differential
activation of cis-acting transcription factors in different
cell types may allow for tissue- and cell-specific patterns of
chemokine expression. This observation suggests that selective
inhibition of the RANTES promoter in particular tissues may be
possible.
The results in the present study explain how IL-1ß activates a
requisite response element allowing the up-regulation of RANTES
expression in ESC. We propose that such a regulatory mechanism
critically influences the local recruitment of leukocytes during the
inflammatory response. Selective interference of the NF-B-induced
transcriptional regulation of RANTES in ESC could lead to the
development of safe treatments to blunt the inflammatory response in
women with endometriosis.
Acknowledgments
We thank the clinical staff of the University of California-San Francisco, Department of Obstetrics, Gynecology, and Reproductive Sciences, for their generous contributions to the study.
Footnotes
This work was supported by the following NIH grants and fellowships: HD-08517 (to D.I.L.) and HD-37321 (to D.I.L. and R.N.T.), through the Specialized Cooperative Centers Program in Reproductive Research.
Abbreviations: ESC, Endometriotic stromal cell(s); GAPDH,
glyceraldehyde-3-phosphate dehydrogenase; IB, inhibitors of B;
NF-B, nuclear factor-B; RANTES, regulated on activation,
normal T cell-expressed and secreted.
Received February 14, 2001.
Accepted June 11, 2001.
References
, LPS, and IL-1 ß. Science 243:1467–1469B regulation by IB kinase in primary
fibroblast-like synoviocytes. J Immunol 163:427–433,
and IL-2 in peritoneal fluid and macrophage-conditioned media of women
with endometriosis. Am J Reprod Immunol 34:381–385
B DNA
binding activity. Biochem Biophys Res Commun 220:120–124[CrossRef][Medline]
B nuclear
binding and is inhibited by adenovirus-mediated expression of inhibitor
of B. J Immunol 161:1007–1016 by lower
airway epithelial cells and eosinophils infected with respiratory
syncytial virus. J Virol 72:4756–4764
B in the immune system. Annu Rev Immunol 12:141–179[Medline]
B. Int J
Biochem Cell Biol 29:867–870[CrossRef][Medline]
B in cytokine gene regulation. Am J Respir Cell Mol Biol 17:3–9B potently up-regulates the promoter activity of RANTES, a
chemokine that blocks HIV infection. J Immunol 158:3483–3491[Abstract]
B and IFN-regulatory factor transcription factors. J
Immunol 164:5352–5361B. J Leukocyte Biol 65:291–298[Abstract]This article has been cited by other articles:
 |
|
 |
|
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|
|
|
|
 |
|
 S Guay and A Akoum Stable inhibition of interleukin 1 receptor type II in Ishikawa cells augments secretion of matrix metalloproteinases: possible role in endometriosis pathophysiology Reproduction, September 1, 2007; 134(3): 525 - 534. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. D. Wren, Y. Wu, and S.-W. Guo A system-wide analysis of differentially expressed genes in ectopic and eutopic endometrium Hum. Reprod., August 1, 2007; 22(8): 2093 - 2102. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. Gonzalez-Ramos, J. Donnez, S. Defrere, I. Leclercq, J. Squifflet, J.-C. Lousse, and A. Van Langendonckt Nuclear factor-kappa B is constitutively activated in peritoneal endometriosis Mol. Hum. Reprod., July 1, 2007; 13(7): 503 - 509. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
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|
|
|
|
|
|
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|
|
|
|
 |
|
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|
|
|
|
|
|
Y. Hirota, Y. Osuga, T. Hirata, O. Yoshino, K. Koga, M. Harada, C. Morimoto, E. Nose, T. Yano, O. Tsutsumi, et al. Possible Involvement of Thrombin/Protease-Activated Receptor 1 System in the Pathogenesis of Endometriosis J. Clin. Endocrinol. Metab., June 1, 2005; 90(6): 3673 - 3679. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
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|
|
|
|
|
|
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|
|
|
|
|
|
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|
|
|
|
|
|
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|
|
|
|
 |
|
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|
|
|
|
 |
|
 S.-M. Huang and D. J. McCance Down Regulation of the Interleukin-8 Promoter by Human Papillomavirus Type 16 E6 and E7 through Effects on CREB Binding Protein/p300 and P/CAF J. Virol., July 29, 2002; 76(17): 8710 - 8721. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
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