Published online before print July 7, 2004
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Kennedy Institute of Rheumatology Division, Imperial College London, Hammersmith, United Kingdom
1 Correspondence: Kennedy Institute of Rheumatology, Imperial College of Science, Technology and Medicine, Charing Cross Campus, ARC Building, 1 Aspenlea Road, Hammersmith, London, W6 8LH, UK. E-mail: brian.foxwell{at}imperial.ac.uk
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Key Words: monocyte/macrophage • signal transduction • inflammation
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TOPABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
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(TNF-) [5
]. The mechanisms by which IL-10 inhibits TNF- gene expression remain unresolved but may include effects on transcription (possibly by the inhibition of nuclear factor-
B activation) and/or effects on the processing of TNF- mRNA via the 3'-untranslated region [6
7
8
]. IL-10-mediated inhibition of p38 mitogen-activated protein kinase (MAPK) activity, which is involved in the post-translational processing of TNF- mRNA, has also been described, although this last mechanism remains controversial [8
, 9
]. Other studies have intimated that de novo protein synthesis is required for the IL-10-mediated suppression of TNF- [10
, 11
], and there is evidence from us and others that STAT-3 is required for this protein synthesis [12
]. Within this context, Lee and Chau [13
] have suggested a role for heme oxygenase-1 (HO-1) in the anti-inflammatory activities of IL-10 in the murine macrophage cell line J774.1 and primary murine peritoneal macrophages.
HO-1 catalyzes the rate-limiting step in the degradation of heme by the oxidative cleavage of the -meso carbon bridge to produce equimolar quantities of Biliverdin-IX, carbon monoxide (CO), and free iron. CO has been shown to have anti-inflammatory activity [14
, 15
]. Moreover the generation of HO-1-deficient mice has also suggested an anti-inflammatory role for HO-1, as these mice display an increased inflammatory state, and transfection of HO-1 into the lungs of rats mediated potent, anti-inflammatory effects [16
17
18
19
]. The anti-inflammatory effects of HO-1 would therefore make it a potential mediator of IL-10 function. However, HO-1-deficient mice do not share the same phenotype as the IL-10 knockout, which develops chronic enterocolitis and dysregulation of inflammatory responses [20
, 21
]. Also, expression of HO-1 can be induced by a wide range of stimuli associated with oxidative stress and has also been found to be induced by nitric oxide, cyclic adenosine monophosphate, hypothermia, hyperthermia, and proinflammatory cytokines (TNF-, IL-1, IL-6), as well as lipopolysaccharide (LPS). In addition, the induction of HO-1 has been found to be mediated by IL-10 via the activation of p38 MAPK [13
], an event not previously associated with the function of the cytokine [15
, 22
23
24
].
This study has investigated the regulation of HO-1 by IL-10 in murine and human monocytes/macrophages and examined how this might correlate with the anti-inflammatory activity of the cytokine. In addition, we have compared the regulation of HO-1 by IL-10 with IL-6, a cytokine known to induce STAT-3 activation and the proinflammatory stimuli LPS. The data presented confirm the previous observations that IL-10 induces the expression of HO-1 in murine macrophages and demonstrates similar activity in human macrophages. Additionally, IL-6 was able to induce HO-1 expression. We find that expression of HO-1 requires the activation of STAT-3 and PI-3K. In contrast, there appears to be no role for p38 MAPK. However, despite this regulation by IL-10, we find no correlation between the expression and activity of HO-1 and the anti-inflammatory effects of the cytokine. Finally, this study also demonstrates that LPS antagonizes HO-1 expression induced by IL-10 in primary human cells, the first time to our knowledge that regulation of IL-10 function by LPS has been observed.
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TOPABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
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RAW264.7 and J774.1 cells were cultured in 10% HIFCS Dulbecco’s modified Eagle’s medium (DMEM) at 37°C, 5% CO2, subculturing cells when 60–70% confluent. RAW264.7 cells were incubated for a minimum of 4 h in 5% HIFCS DMEM media at 37°C, 5% CO2, prior to stimulation. J774 cells were incubated for a minimum of 4 h in 0.1% HIFCS DMEM media at 37°C, 5% CO2, prior to stimulation.
Materials
Anti-ß-actin antibody was from Sigma (Dorset, UK); anti-HO-1 antibody was from Bioquote Ltd. (York, UK); zinc (II) protoporphyrin IX (ZnPp) was from Alexis Biochemicals (Little Chalfont, UK) and Sigma, stored at –20°C in the dark; SB230580 and LY294002 were from Calbiochem (Nottingham, UK); LPS was from Salmonellatyphi from Sigma; IL-10 was kindly donated by Schering Plough (Bloomfield, NJ); M-CSF was a kind donation from Wyeth; and human TNF- was received as a kind donation from Dr. Zehra Kaymakcalan (BASF Corporation, Wyandotte, MI).
Western blot analysis
Whole cell lysates were prepared from adherent cells by washing cell monolayer with phosphate-buffered saline (PBS) and lysed for 15–30 min at 4°C in lysis buffer [1% Triton X-100, 10 mM Tris, pH 7.6, 150 mM NaCl, 1 mM EDTA, 1 µM NaVO3, 2 µg/ml aprotinin, 1 µM 4-(2-aminoethyl) benzenesulfonyl fluoride]. Monocytes were centrifuged at 14,000 g, and cell pellets were lysed for 15–30 min at 4°C in lysis buffer. Lysed samples were centrifuged at 14,000 g for 10 min at 4°C, and the appropriate volume of 5x gel sample buffer [1 M Tris-HCl, pH 6.8, 10% w/v sodium dodecyl sulfate (SDS), 50% v/v glycerol, 12.5% v/v ß-mercaptoethanol, bromophenol blue] was added to the supernatant. Samples were boiled for 5 min at 95°C to denature proteins, resolved by 12% SDS-polyacrylamide gel electrophoresis, and transferred to a polyvinyl-difluoride membrane (Millipore, Bedford, MA). Membranes were blocked with blocking buffer (5% marvel: 0.1% Tween-20 in PBS) and incubated with primary antibody (1:1000) in blocking buffer for a minimum of 1 h at 20°C. Membranes were incubated with anti-rabbit or anti-mouse peroxidase-conjugated polyclonal antibodies (Amersham Biosciences, Little Chalfont, UK) in 5% marvel: 0.1% Tween-20 in PBS (1:2000) for a minimum of 1 h. The membranes were treated with enhanced chemiluminescence (Amersham Biosciences) and were visualized using Hyperfilm MP (Amersham Pharmacia Biotech, Little Chalfont, UK).
Measurement of TNF--soluble p75 TNF receptor (TNF-R) and IL-6 production
TNF--soluble p75 TNF-R and IL-6 concentrations were measured in serum by enzyme-linked immunosorbent assay (ELISA; BD Biosciences, Oxford, UK) according to the manufacturer’s instructions, and absorbance was read at A450 nm.
Infection of in vitro-differentiated (IVD) macrophages by recombinant adenoviruses
Recombinant replication-deficient adenoviral vectors encoding the human STAT-3 Tyr-705-Phe (Ad-STAT-3-DN) were provided by Yasushi Fasjio (University of Osaka, Japan); adenoviral vectors having no insert (Ad
) were provided by Andrew Byrnes and Matthew Wood (University of Oxford, UK). Adenovirus was purified and concentrated as described previously [25
]. Macrophages were exposed to virus at stated multiplicity of infection (MOI) for 1 h in serum-free RPMI followed by culture in 5% HIFCS RPMI for 24 h.
Flow cytometric analysis of human monocytes
Primary human monocytes blocked for 30 min in 1% HI human serum/PBS. Cells were stained with the appropriate fluorochrome-conjugated antibody or fluorochrome-conjugated isotype-control antibody from BD Biosciences for a minimum of 30 min, washed twice with PBS, and resuspended in PBS. Samples were analyzed by FACScan (Becton Dickinson).
Analysis of data
Data are shown as mean ± SD and are representative of at least three experiments.
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Figure 1. IL-10 and LPS regulate the expression of HO-1. (A) Cells of the murine macrophage cell line J774.1 were stimulated with IL-10 (10 ng/ml) over 48 h. (B) The murine macrophage cell line RAW264.7 was left unstimulated or stimulated with IL-10 (10 ng/ml), LPS (1 ng/ml), or both for 6 or 24 h. (C) RAW264.7 cells were preincubated for 30 min with anti-IL-10 receptor (IL-10R) antibody or an isotype control before stimulation with LPS (1 ng/ml) or IL-10 (10 ng/ml). (D) Peritoneal macrophages (M ) were elicited and separated by adherence from 8-week-old, male BALB/c mice. Cells were left unstimulated or stimulated for 24 h with IL-10 (10 ng/ml), LPS (1 ng/ml), or costimulated with IL-10 (10 ng/ml) and LPS (1 ng/ml). Cells were also stimulated with ZnPp (1 µM). (E) Primary human monocytes were left unstimulated or stimulated with IL-10 (10 ng/ml), LPS (1 ng/ml), or both for 6 or 24 h. (F) Primary human macrophages were left unstimulated (uns) or stimulated over 24 h with LPS (1 ng/ml) ± IL-10 (10 ng/ml) or IL-6 (20 ng/ml), and the soluble TNF- was measured by ELISA. (G) Human macrophages were stimulated for 6 or 24 h with IL-6 (10 ng/ml), LPS (1 ng/ml), and IL-10 (10 ng/ml) as indicated. In all cases, whole cell lysates were prepared and analyzed for HO-1 expression by Western blot. In each study, the blots were reprobed with ß-actin as a loading control. These data are representative of at least three experiments.
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To ascertain if the responses of the J774.1 and RAW264.7 cell lines are representative of responses in primary murine peritoneal macrophages, the studies were repeated in these cells. Peritoneal macrophages were stimulated with IL-10 and/or LPS for 24 h. These data demonstrate that in isolation, IL-10 and LPS induce the expression of HO-1. However, in contrast to RAW264.7 and J774.1 cells, costimulation results in a reduced level of HO-1 expression, compared with either activator alone (Fig. 1D) .
IL-10 but not LPS induces HO-1 expression in human monocytes
To determine the regulation of HO-1 production in a more physiologically relevant system, HO-1 expression was evaluated in primary human monocytes. It has previously been shown that HO-1 is induced by IL-10 in adherent monocytes but not nonadherent monocytes [27
]. We have comfirmed the expression of IL-10-induced HO-1 in human monocytes, which were stimulated for 6 or 24 h in the presence of IL-10 or LPS. Little HO-1 expression was induced by IL-10 by 6 h, but the protein was clearly seen by 24 h (Fig. 1E) . However, when stimulated with LPS, there was no increase in protein expression, and costimulation resulted in a reduced IL-10 response. Similar results were obtained in human IVD macrophages (data not shown).
IL-6 also induces HO-1 expression
IL-6 demonstrates some inhibitory effect on TNF- expression (Fig. 1F)
; however, this effect is considerably lower than that of IL-10. IL-6 shares common signaling mechanisms with IL-10, such as the activation of Jak kinases and STAT-3. We therefore examined if IL-6 was capable of inducing HO-1 expression. In J774.1 cells, IL-6 induced HO-1 expression very effectively by 24 h (data not shown); similar results were obtained in RAW264.7 cells (data not shown). Human macrophages, when stimulated with IL-6 for 6 or 24 h, also expressed HO-1 (Fig. 1G)
.
In macrophages, IL-10-induced HO-1 is not mediated by p38 MAPK
The previous study by Lee and Chau [13
] suggested that the mechanism by which IL-10 induces HO-1 expression involves the p38 MAPK pathway; however, MAPKs have been demonstrated to have different effects on HO-1 expression depending on the cell line and in some cases, the conditions of stimulation [13
, 28
]. Therefore, SB230580, an inhibitor of p38 MAPK, was used to examine the role of the kinase in the signaling process used by IL-10 or LPS to modulate the expression of HO-1.
In RAW264.7 cells, inhibition of p38 MAPK with SB230580 results in the ablation of LPS-induced HO-1 expression, and IL-10-induced expression remains largely unaffected (Fig. 2A
). In human macrophages, SB203580 had no effect on IL-10-induced HO-1 expression or the suppression of expression displayed by LPS (Fig. 2B)
. To further examine the role of p38 MAPK in the human system, a dominant-negative form of the kinase, p38 MAPK-D168A, was introduced into macrophages by an adenoviral vector (Ad-p38-DN). We have previously found this dominant-negative p38 MAPK to be a potent inhibitor of LPS-induced TNF- production (manuscript in preparation). As shown in Figure 2C
, cells that were infected with Ad-p38-DN show no alteration in HO-1 expression, demonstrating that in this human system, the kinase is not required by IL-10. Unfortunately, we were unable to obtain expression of p38-DN protein on viral infection of the RAW264.7 cell line.
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Figure 2. Inhibition of p38 MAPK abrogates LPS-induced HO-1 in RAW264.7 cells but has no discernible effect in human IVD macrophages. (A) RAW264.7 cells or (B) human macrophages (M) were treated with SB203580 (2 µM) or were left untreated, followed by stimulation for 24 h with IL-10 (10 ng/ml) or LPS (1 ng/ml). Human macrophages were also costimulated with IL-10 (10 ng/ml) and LPS (1 ng/ml). Whole cell lysates were analyzed by Western blot for HO-1 expression. (C) Human macrophages were left uninfected or infected with Ad or Ad-p38-DN (MOI=200) as described in Materials and Methods. The cells where then treated with IL-10 (10 ng/ml) and/or LPS (1 ng/ml) for 24 h. Whole cell lysates were analyzed for HO-1 expression by Western blot. In each study, the blots were reprobed with ß-actin as a loading control. These data are representative of at least three experiments.
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Figure 3. IL-10-stimulated HO-1 expression is dependent on STAT-3 in IVD macrophages. Human macrophages were left uninfected or infected with Ad or Ad-STAT-3-DN (MOI=200) as described in Materials and Methods. The cells were treated with IL-10 (10 ng/ml) for 24 h, after which, whole cell lysates were analyzed for HO-1 expression by Western blot. The blot was reprobed with STAT-3. These data are representative of at least three experiments.
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by IL-10. However, Figure 4C
demonstrates that inhibition of PI-3K by LY294002 does not significantly affect the inhibition of TNF- production by IL-10.
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Figure 4. PI-3K is required for IL-10-induced expression of HO-1 in macrophages (M). (A) Human IVD macrophages or (B) RAW264.7 cells were treated with LY294002 (10 µM) or were left untreated, followed by stimulation for 24 h with IL-10 (10 ng/ml) and/or LPS (1 ng/ml). Whole cell lysates were prepared and analyzed by Western blot for HO-1 expression. In each study, the blots were reprobed with ß-actin as a loading control. (C) Human IVD macrophages were pretreated with LY294002 and then stimulated with 1 ng/ml LPS (open bars) ± 10 ng/ml IL-10 (solid bars) for 6 h, and the TNF- levels in the supernatants were analyzed by ELISA. These data are representative of at least three experiments. uns, Unstimulated.
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expression. In primary human monocytes (Fig. 6A
), human IVD macrophage, or the mouse cell line RAW264.7 cells (data not shown), LPS-induced TNF- levels are unaffected by ZnPp inhibition of HO-1 activity nor did the drug have any effect on inhibition of TNF- production by IL-10. Therefore, we would conclude that HO-1 activity does not play any significant role in the suppression of TNF- induction by IL-10. This study was then extended to a second cytokine known to be suppressed by IL-10, IL-6, the LPS-induced expression of which in the absence or presence of IL-10, is also unaffected by the inhibition of HO-1 (Fig. 6B)
. In addition to the direct regulation of proinflammatory cytokine production, IL-10 is known to have other effects on inflammation, for example, the secretion of soluble TNF-R (Sp75-TNF-R) and the modulation of costimulatory molecule expression. The treatment of cells with ZnPp had no effect on Sp75-TNF-R production induced by IL-10 (Fig. 6C)
. The effects of HO-1 on IL-10-mediated regulation of human leukocyte antigen (HLA) molecules and the costimulatory molecule CD86 were assessed by fluorescein-activated cell sorter analysis after stimulation with IL-10 ± ZnPp. In this study, inhibition of HO-1 had no effect on IL-10-mediated regulation of HLA-DR molecules (Fig. 6D)
or on costimulatory CD86 molecules (data not shown).
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Figure 5. Stimulation of human myeloid cells with ZnPp. Human macrophages were stimulated with the HO-1 inhibitor ZnPp (1 µM) with dimethylformamide (DMF) as vehicle control. Whole cell lysates were prepared and analyzed by Western blot for HO-1 expression. In each study, the blots were reprobed with ß-actin as a loading control. uns, Unstimulated.
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Figure 6. HO-1 is independent of the anti-inflammatory effects of IL-10. Human macrophages were preincubated with ZnPp at a range of concentrations (1–0.008 µM) followed by stimulation with LPS (1 ng/ml) ± IL-10 (10 ng/ml) for 24 h. (A) TNF- and (B) IL-6 concentrations in the supernatants were analyzed by ELISA. (C) Human macrophages were preincubated with ZnPp (1 µM) or vehicle control (DMF) for 30 min. Cells were left unstimulated or stimulated with IL-10 (10 ng/ml) for 24 h. The concentration of soluble TNF-R (Sp75) was determined by ELISA. (D) Monocytes treated in the same manner were stained with the appropriate fluorochrome-conjugated antibody (HLA-DR: PE) or fluorochrome-conjugated isotype-control antibody. These data are representative of at least three experiments.
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TOPABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
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production within 1–2 h of stimulation with LPS [8
, 12
]. Third, IL-6 strongly induces HO-1 at 6 h but does not display a similar level of anti-inflammatory activity to IL-10 at the time of HO-1 induction. Fourthly, in the murine macrophage cell line RAW264.7, LPS induced HO-1 expression more potently than IL-10. Finally, and most importantly, specific inhibition of HO-1 activity did not affect IL-10 anti-inflammatory activity regardless of the fact that the macrophages contain very high levels of the protein. Our data, although in dispute with those of Lee and Chau [13 ], do agree with those from HO-1-deficient mice, which display an inflammatory phenotype but do not reproduce the phenotype of spontaneous inflammatory bowel syndrome seen in IL-10-deficient mice [18 , 21 ]. Furthermore, we were unable to show any role for p38 MAPK in IL-10-induced HO-1 expression, a result obtained with the kinase inhibitor SB203580 as well as a dominant-negative inhibitor of p38 MAPK. Our results are in agreement with studies that have described this kinase as a target of IL-10 anti-inflammatory function [9 ] or having no relationship at all [8 ]. In contrast, in murine cells, p38 MAPK was required for LPS-induced expression of HO-1. A possible explanation for the differences in the data concerning p38 MAPK could be a result of the concentration of the p38 MAPK inhibitor used. In the present study, 2 µM SB230580 was used, a concentration adequate to block LPS-induced expression of HO-1, whereas Lee and Chau [13 ] used 10 µM. At such high concentrations, SB230580 can inhibit the PI-3K pathway by blocking phosphatidyl-dependent kinase 1 and 2 [32 ]. As this pathway is required by IL-10 to induce HO-1, this could provide an explanation for their data.
We have previously shown that IL-10 induces PI-3K activity [4 ]. However, we were previously unable to assign any biological function to this activity in human monocytes. The observation that IL-10-induced PI-3K is required for HO-1 expression is the first time, to our knowledge, that a biological response has been associated for IL-10-induced PI-3K within macrophages. Our studies show that STAT-3 is also required for expression of HO-1, suggesting that the two pathways may work in concert to drive expression. Exactly how the two pathways may relate is beyond the present study.
Less straightforward was the regulation of HO-1 by LPS, which was able to induce HO-1 expression in murine RAW264.7 and primary murine macrophages and required the activation of p38 MAPK and PI-3K. In RAW264.7 cells, HO-1 expression was enhanced by costimulation with IL-10, and in primary murine cells, expression was reduced by costimulation. However, the LPS induction of HO-1 expression could not be reproduced in human myeloid cells, where LPS inhibited the IL-10- and the IL-6-induced HO-1. Such a dichotomy between HO-1 expression in human and murine systems has been observed with various other stimuli such as interferon-
[33
], where the stimulus up-regulates HO-1 in the murine system but not in the human. However, this is the first time in a primary human system that LPS signaling has been shown to interfere with the effects of IL-10. This observation demonstrates that human and murine cells can behave differently and may be an explanation for the diversity of mechanisms ascribed to IL-10; i.e., they may result from species-specific differences.
Overall, given the promiscuity of stimuli that can elicit the expression of HO-1, whose function is to reduce oxidative stress, it is difficult to see how it could be pivotal to the anti-inflammatory activity of IL-10. The suppression of HO-1 expression by LPS seen in human cells could have an important rationale. It has been postulated by Shibahara et al. [34 ] that HO-1 is highly energy-dependent, requiring colocalization with the cytochrome P450 reductase, which provides 1 mole of nicotinamide adenine dinucleotide phosphate per mole of heme catabolized. This may be prohibitive for cells involved in fighting infection [34 ]. Therefore, stimulation by LPS, a component of bacterial cell membranes, could potentially signal a repression of HO-1 to conserve energy. Although IL-10, produced as a negative regulator post-infection, may stimulate HO-1 expression as a sink for reactive oxygen species produced during infection. However, this is conjecture, and the physiological role in human cells for IL-10-induced HO-1 remains to be fully elucidated. Furthermore, if this hypothesis is correct, it is difficult to understand why human and murine cells behave so differently.
In summary, this study has investigated the regulation of HO-1 expression by IL-10 and LPS. Although IL-10 clearly regulates HO-1 expression in human and murine myeloid cells, there is little evidence to support any major role for this event in the anti-inflammatory activity of the cytokine. The study shows that STAT-3 and PI-3K pathways are required for the induction of HO-1 by IL-10, but we find no evidence of a role for p38 MAPK. The regulation of HO-1 expression by LPS showed a clear species disparity with induction in murine cells and inhibition in human cells. It is difficult to understand at present why HO-1 regulation may need to differ so much, but the data do demonstrate clearly that gene regulation cannot be assumed to be identical across species barriers.
Received January 28, 2004; revised May 17, 2004; accepted May 27, 2004.
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STAT3 complexes in human T cells and monocytes J. Immunol. 155,1079-1090[Abstract]B (NFB) activation in human monocytes: IL-10 and IL-4 suppress cytokine synthesis by different mechanisms J. Biol. Chem. 270,9558-9563B activity in murine monocytes/macrophages DNA Cell Biol. 22,131-139[CrossRef][Medline] production that does not involve inhibition of p38 mitogen-activated protein kinase or NF- B in primary human macrophages J. Immunol. 168,4837-4845B reveals that macrophage tumor necrosis factor production in rheumatoid arthritis is NF-B dependent Proc. Natl. Acad. Sci. USA 95,8211-8215 in human glioblastoma cells J. Neurochem. 72,2356-2361[CrossRef][Medline]This article has been cited by other articles:
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|
|
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