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• W2039226089 abstract "Annexin 1 (ANXA1) is a calcium-binding protein endowed with anti-inflammatory properties. Using an extra-hepatic system, we showed that interleukin (IL)-6 regulates ANXA1 expression at the transcriptional level. The purpose of this study was to determine whether ANXA1 synthesis was modulated by IL-6 during experimental inflammation. We have compared liver ANXA1 expression during systemic and localized inflammatory reaction, using lipopolysaccharide (LPS) and turpentine. LPS treatment strongly induced ANXA1 expression in the liver of wild-type (WT) animals (+600%) whereas a modest increase (+60%) was measured in IL-6 knockout (KO) animals. Turpentine treatment did not affect the expression of ANXA1 in either animal type. LPS enhanced serum corticosteroid levels equally in WT and IL-6 KO mice, whereas higher tumor necrosis factor (TNF)-α and IL-1β levels were released in IL-6 KO animals. Injection of mouse recombinant IL-6 to IL-6 KO animals before LPS or TNF-α challenge, replenished ANXA1 liver synthesis to that of WT animals. Exogenous ANXA1 but not ANXA5, administered to IL-6 KO mice before LPS challenge inhibited TNF-α release. We propose that ANXA1 acts as a novel acute phase protein, which is controlled in the liver by TNF-α and IL-6, and which may contribute to the resolution of systemic endotoxemia through a negative feedback on TNF-α release. Annexin 1 (ANXA1) is a calcium-binding protein endowed with anti-inflammatory properties. Using an extra-hepatic system, we showed that interleukin (IL)-6 regulates ANXA1 expression at the transcriptional level. The purpose of this study was to determine whether ANXA1 synthesis was modulated by IL-6 during experimental inflammation. We have compared liver ANXA1 expression during systemic and localized inflammatory reaction, using lipopolysaccharide (LPS) and turpentine. LPS treatment strongly induced ANXA1 expression in the liver of wild-type (WT) animals (+600%) whereas a modest increase (+60%) was measured in IL-6 knockout (KO) animals. Turpentine treatment did not affect the expression of ANXA1 in either animal type. LPS enhanced serum corticosteroid levels equally in WT and IL-6 KO mice, whereas higher tumor necrosis factor (TNF)-α and IL-1β levels were released in IL-6 KO animals. Injection of mouse recombinant IL-6 to IL-6 KO animals before LPS or TNF-α challenge, replenished ANXA1 liver synthesis to that of WT animals. Exogenous ANXA1 but not ANXA5, administered to IL-6 KO mice before LPS challenge inhibited TNF-α release. We propose that ANXA1 acts as a novel acute phase protein, which is controlled in the liver by TNF-α and IL-6, and which may contribute to the resolution of systemic endotoxemia through a negative feedback on TNF-α release. The synthesis of acute phase proteins (APPs) is a protective mechanism of the host during conditions of distress, inflammation, and systemic endotoxemia.1Shumann R Kirschning C Unbehaun A Aberle H Knopf H-P Lamping N Ulevitch RJ Herrmann F The lipopolysaccharide-binding protein is a secretory class 1 acute-phase protein whose gene is transcriptionally activated by APRF/STAT-3 and other cytokine-inducible nuclear proteins.Mol Cell Biol. 1996; 16: 3490-3503PubMed Google Scholar The synthesis and release of class 1 APP (eg, serum amyloid A) is initiated by the proinflammatory cytokines interleukin (IL)-1 and tumor necrosis factor (TNF)-α via a mechanism that is IL-6 mediated. Class 2 APPs (eg, α2-macroglobulin) are induced by IL-6 alone and by cytokines related to the IL-6 family.1Shumann R Kirschning C Unbehaun A Aberle H Knopf H-P Lamping N Ulevitch RJ Herrmann F The lipopolysaccharide-binding protein is a secretory class 1 acute-phase protein whose gene is transcriptionally activated by APRF/STAT-3 and other cytokine-inducible nuclear proteins.Mol Cell Biol. 1996; 16: 3490-3503PubMed Google Scholar APPs provide a buffering mechanism that protects the organism from the damaging actions of proinflammatory mediators that are produced in large amounts during acute and chronic inflammatory diseases.2Baumann H Gauldie J The acute phase response.Immunol Today. 1994; 15: 74-80Abstract Full Text PDF PubMed Scopus (428) Google Scholar, 3Stadnyk A Gauldie J The acute phase protein response during parasitic infection.Immunol Today. 1991; 12: A7-A12Abstract Full Text PDF PubMed Scopus (54) Google Scholar, 4Gabay C Kushner I Acute-phase proteins and other systemic responses to inflammation.N Engl J Med. 1999; 340: 448-454Crossref PubMed Scopus (4949) Google Scholar The role of endogenous IL-6 in regulating class 2 rather than class 1 APPs has been also addressed using mice deficient in the IL-6 gene [IL-6 knockout (KO) mice].5Fattori E Cappelletti M Costa P Sellitto C Cantoni L Carelli M Faggioni R Fantuzzi G Ghezzi P Poli V Defective inflammatory response in interleukin 6-deficient mice.J Exp Med. 1994; 180: 1243-1250Crossref PubMed Scopus (487) Google Scholar In these animals, the hepatic expression of α2-macroglobulin is compromised after intramuscular injection of turpentine oil,6Kushner I Regulation of the acute phase response by cytokines.Perspect Biol Med. 1993; 36: 611-622Crossref PubMed Scopus (230) Google Scholar whereas the acute-phase response induced by lipopolysaccharide (LPS) is unchanged. The contribution of other cytokines such as TNF-α, which is released in higher amounts in the IL-6 KO mice, has been implicated in this response.5Fattori E Cappelletti M Costa P Sellitto C Cantoni L Carelli M Faggioni R Fantuzzi G Ghezzi P Poli V Defective inflammatory response in interleukin 6-deficient mice.J Exp Med. 1994; 180: 1243-1250Crossref PubMed Scopus (487) Google Scholar Annexin 1 (ANXA1) is a glucocorticoid-inducible protein endowed with profound anti-inflammatory activity. Its powerful action in experimental models of inflammation were attributed originally to its ability to inhibit the activity of phospholipase A2 and, therefore, the production of eicosanoids.7Blackwell GJ Carnuccio R Di Rosa M Flower RJ Parente L Persico P Macrocortin: a polypeptide causing the anti-phospholipase effect of glucocorticoids.Nature. 1980; 287: 147-149Crossref PubMed Scopus (692) Google Scholar, 8Russo-Marie F Duval D Dexamethasone-induced inhibition of prostaglandin production dose not result from a direct action on phospholipase activities but is mediated through a steroid-inducible factor.Biochim Biophys Acta. 1982; 712: 177-185Crossref PubMed Scopus (73) Google Scholar More recent studies have demonstrated ANXA1 ability to down-regulate the process of neutrophil9Perretti M Croxtall JD Wheller SK Goulding NJ Hannon R Flower RJ Mobilizing lipocortin 1 in adherent human leukocytes downregulates their transmigration.Nat Med. 1996; 2: 1259-1262Crossref PubMed Scopus (194) Google Scholar or monocyte10Solito E Romero IA Marullo S Russo-Marie F Weksler BB Annexin1 binds to U937 monocytic cells and inhibits their adhesion to microvascular endothelium: involvement of the α4β1 integrin.J Immunol. 2000; 165: 1573-1581PubMed Google Scholar adhesion to the activated endothelium. Our previous studies using the lung adenocarcinoma A549 cell line showed that ANXA1 expression is up-regulated by IL-6 and corticosteroids.11Solito E de Coupade C Parente L Flower R Russo Marie F IL-6 stimulates Annexin 1 expression and translocation and suggests a new biological role as class II acute phase protein.Cytokine. 1998; 10: 514-521Crossref PubMed Scopus (70) Google Scholar The response to IL-6 was mediated by a C/EBP β transcriptional factor that binds to a specific region of 30 bp.12Solito E de Coupade C Parente L Flower RJ Russo-Marie F Human annexin 1 is highly expressed during the differentiation of the human epithelial cell line A 549. Involvement of NF-IL6 in phorbol ester induction of annexin 1.Cell Growth Differ. 1998; 9: 327-336PubMed Google Scholar From the pattern of stimulation induced by IL-6 and dexamethasone we have proposed that ANXA1 may participate in host defense as a new APP.11Solito E de Coupade C Parente L Flower R Russo Marie F IL-6 stimulates Annexin 1 expression and translocation and suggests a new biological role as class II acute phase protein.Cytokine. 1998; 10: 514-521Crossref PubMed Scopus (70) Google Scholar ANXA1 is expressed in a tissue-specific manner in rodents and, for instance, the liver13Fava RA McKanna J Cohen S Lipocortin 1 (p35) is abundant in a restricted number of differentiated cell types in adult organs.J Cell Physiol. 1989; 141: 284-293Crossref PubMed Scopus (76) Google Scholar or primary hepatic cell14de Coupade C Gillet R Bennoun M Briand P Russo-Marie F Solito E Annexin 1 expression and phosphorylation are upregulated during liver regeneration and transformation in antithrombin III SV40 T large antigen transgenic mice.Hepatology. 2000; 31: 371-380Crossref PubMed Scopus (89) Google Scholar show negligible expression of ANXA1 in basal conditions. However, transgenic mice that developed a hepatocarcinoma expressed ANXA1 (in the liver) in a strictly temporal manner, ie, before tumor development. Similarly, up-regulation of an ANXA1 isoform phosphorylated on tyrosine 21 was detected during liver regeneration after partial hepatectomy.14de Coupade C Gillet R Bennoun M Briand P Russo-Marie F Solito E Annexin 1 expression and phosphorylation are upregulated during liver regeneration and transformation in antithrombin III SV40 T large antigen transgenic mice.Hepatology. 2000; 31: 371-380Crossref PubMed Scopus (89) Google Scholar Glucocorticoid hormones modulate several facets of the host inflammatory response. During experimental endotoxemia, circulating glucocorticoid [corticosterone (CCS) in rodents] levels increase in a time-dependent manner.15Zuckerman SH Shellhaas J Butler LD Differential regulation of lipopolysaccharide-induced interleukin 1 and tumor necrosis factor synthesis: effects of endogenous and exogenous glucocorticoids and the role of the pituitary-adrenal axis.Eur J Immunol. 1989; 19: 301-305Crossref PubMed Scopus (302) Google Scholar The role of this hormonal response is not to down-regulate cytokine production,16Perretti M Duncan GS Flower RJ Peers SH Serum corticosterone, interleukin-1 and tumour necrosis factor in rat experimental endotoxaemia: comparison between Lewis and Wistar strains.Br J Pharmacol. 1993; 110: 868-874Crossref PubMed Scopus (55) Google Scholar but rather to favor the synthesis of APP in the liver.2Baumann H Gauldie J The acute phase response.Immunol Today. 1994; 15: 74-80Abstract Full Text PDF PubMed Scopus (428) Google Scholar, 17Xing Z Gauldie J Cox G Baumann H Jordana M Lei XF Achong MK IL-6 is an antiinflammatory cytokine required for controlling local or systemic acute inflammatory responses.J Clin Invest. 1998; 101: 311-320Crossref PubMed Scopus (1161) Google ScholarIn vitro, glucocorticoids are required for optimal APP induction by IL-1 or IL-6.1Shumann R Kirschning C Unbehaun A Aberle H Knopf H-P Lamping N Ulevitch RJ Herrmann F The lipopolysaccharide-binding protein is a secretory class 1 acute-phase protein whose gene is transcriptionally activated by APRF/STAT-3 and other cytokine-inducible nuclear proteins.Mol Cell Biol. 1996; 16: 3490-3503PubMed Google Scholar The present study was performed to monitor ANXA1 expression in the liver of wild-type (WT) and IL-6 KO animals during experimental inflammation. ANXA1 expression was almost absent in the liver of naïve mice. LPS and turpentine were used as agents inducing systemic or local inflammation, respectively. ANXA1 blocks TNF-α release occurring during a systemic inflammatory reaction. We propose ANXA1 as a novel APP, which contributes to the resolution of inflammation. Male C57BL/6J IL6 KO18Kopf M Baumann H Freer G Freudenberg M Lamers M Kishimoto T Zinkernagel R Bluethmann H Kohler G Impaired immune and acute-phase responses in interleukin-6-deficient mice.Nature. 1994; 368: 339-342Crossref PubMed Scopus (1487) Google Scholar (kindly provided by Transgenic Alliance-IFFA-CREDO, Lyon, France) and control mice (C57BL/6J) from the same genetic background (Center d'Elevage R. Janvier, Le Genest St. Isle, France) (28 to 32g body weight) were used for all experiments. Mice were maintained in standard conditions under a 12-hour light/dark cycle and fed ad libitum a chow diet of 6.5% fat, 53% carbohydrate, and 18.6% protein. The procedure followed in the care and killing of the study animals was in accordance with European Community standards on the care and use of laboratory animals (Ministère de l’Agriculture, France; authorization no. 1975). LPS (from Escherichia coli serotype 055:B5) was purchased from Sigma Chemical Co. (St Louis, MO), resuspended in sterile pyrogen-free saline solution and injected intraperitoneally at a dose of 1 mg/kg body weight.19Ajuebor MN Das AM Virag L Flower RJ Szabo C Perretti M Role of resident peritoneal macrophages and mast cells in chemokine production and neutrophil migration in acute inflammation: evidence for an inhibitory loop involving endogenous IL-10.J Immunol. 1999; 162: 1685-1691PubMed Google Scholar A volume of 100 μl of steam-distilled turpentine was injected intramuscularly.5Fattori E Cappelletti M Costa P Sellitto C Cantoni L Carelli M Faggioni R Fantuzzi G Ghezzi P Poli V Defective inflammatory response in interleukin 6-deficient mice.J Exp Med. 1994; 180: 1243-1250Crossref PubMed Scopus (487) Google Scholar Turpentine oil (British Pharmacopoeia) was from Thornton and Ross (Huddersfield, England). Mouse recombinant TNF-α or mouse recombinant IL-6 (R & D Systems, Oxon, UK) were administered intravenously at a dose of 1 μg per mouse.17Xing Z Gauldie J Cox G Baumann H Jordana M Lei XF Achong MK IL-6 is an antiinflammatory cytokine required for controlling local or systemic acute inflammatory responses.J Clin Invest. 1998; 101: 311-320Crossref PubMed Scopus (1161) Google Scholar, 20Tissi L Puliti M Barluzzi R Orefici G von Hunolstein C Bistoni F Role of tumor necrosis factor alpha, interleukin-1beta, and interleukin-6 in a mouse model of group B streptococcal arthritis.Infect Immun. 1999; 67: 4545-4550PubMed Google Scholar, 21Romano M Sironi M Toniatti C Polentarutti N Fruscella P Ghezzi P Faggioni R Luini W van Hinsbergh V Sozzani S Bussolino F Poli V Ciliberto G Mantovani A Role of IL-6 and its soluble receptor in induction of chemokines and leukocyte recruitment.Immunity. 1997; 6: 315-325Abstract Full Text Full Text PDF PubMed Scopus (903) Google Scholar Human recombinant ANXA1 and ANXA5 were administrated intravenously at a dose of 10 μg per animal 30 minutes before LPS treatment.22Lim LH Solito E Russo-Marie F Flower RJ Perretti M Promoting detachment of neutrophils adherent to murine postcapillary venules to control inflammation: effect of lipocortin 1.Proc Natl Acad Sci USA. 1998; 95: 14535-14539Crossref PubMed Scopus (154) Google Scholar Neutralizing rabbit anti-mouse IL-6 or anti-mouse TNF-α polyclonal IgG (R&D Systems, Oxon, UK) were administered intraperitoneally (5 μg per mouse) 1 hour before LPS (1 mg/kg intraperitoneally).23Perretti M Szabo C Thiemermann C Effect of interleukin-4 and interleukin-10 on leucocyte migration and nitric oxide production in the mouse.Br J Pharmacol. 1995; 116: 2251-2257Crossref PubMed Scopus (67) Google Scholar Whereas the glucocorticoid antagonist RU-486 (or mifepristone; Roussel-Uclaf, Romainville, France) was co-injected with LPS, at the dose of 20 mg/kg intraperitoneally.24Perretti M Flower RJ Measurement of lipocortin 1 levels in murine peripheral blood leukocytes by flow cytometry: modulation by glucocorticoids and inflammation.Br J Pharmacol. 1996; 118: 605-610Crossref PubMed Scopus (81) Google Scholar At the reported times after stimulus administration, liver total RNA from control and IL-6 KO mice was extracted using the RNAeasy Qiagen Kit (TEBU, Paris, France), following the manufacturer's instructions. After hybridization with the ANXA1 cDNA-radiolabeled probe, filters were hybridized to a DNA fragment coding for glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as previously reported.25Solito E Raguenes-Nicol C de Coupade C Bisagni-Faure A Russo-Marie F U 937 cells deprived of annexin 1 demonstrate an increased PLA2 activity.Br J Pharmacol. 1998; 124: 1675-1683Crossref PubMed Scopus (45) Google Scholar RNA-DNA hybridization was quantified by densitometric computer analysis in a series 400 Phosphorimager from Molecular Dynamics (Sunnyvale, CA). For protein analysis, liver proteins from WT or KO animals treated or not were extracted in RIPA buffer26Ausubel FM Brent R Kingston R Moore DD Seidman JG Smith JA Struhl K Current Protocols in Molecular Biology. John Wiley & Sons, Inc., Boston1995Google Scholar containing protease and phosphatase inhibitors [100 μmol/L phenylmethyl sulfonyl fluoride, 1 μg/ml leupeptin, 1 μg/ml aprotinin (Boehringer Mannheim, Indianapolis, IN), 1 μmol/L Na3VO4, 1 μmol/L NaF (Boehringer)]. Protein aliquots (30 μg) were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis27Laemmli UK Cleavage of structural proteins during the assembly of the head of bacteriophage T4.Nature. 1970; 227: 680-685Crossref PubMed Scopus (205531) Google Scholar and electroblotted onto nitrocellulose membranes (Bio-Rad, Hercules, CA). Immunoreactive proteins were revealed after immunoblotting with a rabbit polyclonal ANXA1 antibody (1/1000).14de Coupade C Gillet R Bennoun M Briand P Russo-Marie F Solito E Annexin 1 expression and phosphorylation are upregulated during liver regeneration and transformation in antithrombin III SV40 T large antigen transgenic mice.Hepatology. 2000; 31: 371-380Crossref PubMed Scopus (89) Google Scholar A mouse monoclonal α-tubulin antibody (1/1000; Amersham SA, France) was used as internal control for protein level standardization. Densitometric analysis was performed using an Ultrascan XL Laser densitometer (Agfa, Ridgefield Park, NJ). Hepatocytes were isolated by in situ collagenase perfusion of IL-6 KO mice liver according to Decaux and colleagues.28Decaux JF Antoine B Kahn A Regulation of the expression of the L-type pyruvate kinase gene in adult rat hepatocytes in primary culture.J Biol Chem. 1989; 264: 11584-11590Abstract Full Text PDF PubMed Google Scholar Perfused liver was minced in M199 medium (Life Technologies, Inc., Gaithersburg, MD) and filtered through a 70-μm-mesh filter. Viability of recovered cells exceeded 90% as determined by trypan blue exclusion test. Hepatocytes were then plated at a density of 1 × 106 in 60-mm diameter and keep in culture according to Decaux and colleagues,28Decaux JF Antoine B Kahn A Regulation of the expression of the L-type pyruvate kinase gene in adult rat hepatocytes in primary culture.J Biol Chem. 1989; 264: 11584-11590Abstract Full Text PDF PubMed Google Scholar in the presence of 1 μmol/L of dexamethasone (Sigma Chemical Co., St. Louis, MO). After 3 hours, medium was removed and the hepatocytes were cultured under serum-free conditions at 37°C in 5% CO2 atmosphere. Blood CCS levels were measured by radioimmunoassay, according to the manufacturer's instructions (ICN Pharmaceuticals Ltd., Basingstoke, UK). An enzyme-linked immunosorbent assay kit purchased from R&D System (Abingdon, UK) was used to measure serum TNF-α levels. Fresh resected liver and specimens were fixed in 4% paraformaldehyde and embedded in paraffin as described.14de Coupade C Gillet R Bennoun M Briand P Russo-Marie F Solito E Annexin 1 expression and phosphorylation are upregulated during liver regeneration and transformation in antithrombin III SV40 T large antigen transgenic mice.Hepatology. 2000; 31: 371-380Crossref PubMed Scopus (89) Google Scholar Sections (5 μm) were immunostained with a polyclonal antibody directed against the N-terminal domain of ANXA1 (1:5000 final dilution) and subsequently developed using a Vectastain ABC kit (Vector Laboratories, Burlingame, Ca). Preimmune serum was used for control staining.14de Coupade C Gillet R Bennoun M Briand P Russo-Marie F Solito E Annexin 1 expression and phosphorylation are upregulated during liver regeneration and transformation in antithrombin III SV40 T large antigen transgenic mice.Hepatology. 2000; 31: 371-380Crossref PubMed Scopus (89) Google Scholar All values in the figures and text are expressed as mean ± SEM of n observations, where n represents the number of animals studied. Data sets were examined by one- and two-way analysis of variance, and individual group means were compared with Student's unpaired t-test. A P value < 0.05 was considered significant. A low degree of ANXA1 expression was detected in the livers of WT and IL-6 KO mice in basal conditions (Figure 1A). Treatment with LPS (1 mg/kg body weight, intraperitoneally) induced a time-dependent expression of the protein and the mRNA with a peak at 4 hours (approximately sixfold increase greater than basal expression; Figure 1, B and C, referred to the protein level whereas Figure 1D is referred to mRNA expression). ANXA1 protein expression was elevated in WT animals also at the 24-hour time point, whereas it had gone back to basal values by 48 hours after LPS. A modest increase in liver ANXA1 protein level was measured in IL-6 KO mice at all time points under study, with an approximate increase of 60% at 4 hours after LPS (Figure 1B). Similarly, the induction in ANXA1 expression did not last up to the 24-hour time point. Treatment of mice with turpentine did not modify ANXA1 expression in the liver of either WT or IL-6 KO animals (Figure 1C). No differences in lung or thymus ANXA1 were detected between WT and IL-6 KO mice either in the basal condition or after LPS or turpentine treatment (data not shown). Therefore in the next series of experiments we investigated the in vivo mechanism(s) operating during endotoxemia to increase ANXA1 expression in the liver. To explore whether CCS could participate in ANXA1 induction after LPS administration, we measured serum CCS level both before and after treatment with the two inflammogens. Figure 2 A shows that equivalent CCS values were measured in WT and IL-6 KO mice under basal conditions. A comparable increase in CCS plasma levels was seen after LPS treatment in both animal types, with a peak at 4 hours after LPS and values back to normal by the 24-hour time point. LPS treatment induced a significant IL-1β release maximal at 4 hours in both animal types with a higher magnitude in IL-6 KO mice (∼3 times more than WT mice; Figure 2B). A similar sharp increase in serum TNF-α concentration was measured after LPS injection: in IL-6 KO mice TNF-α levels were almost twice as high as those measured in WT mice. After the 90-minute time point, a time-dependent decrease was observed in both animal species, returning almost back to basal by the 4-hour time point (Figure 2C). To identify the cellular source of ANXA1 in the liver, immunohistological analysis was performed on tissues collected from WT and IL-6 KO animals, with or without treatment with LPS. Control liver tissues from WT and KO animals show no obvious staining for ANXA1 (Figure 3, a and b). The large majority of ANXA1 expression in the liver of endotoxic mice was observed in most hepatocytic cells 4 hours after LPS injection into WT mice (Figure 3c), and with less expression in IL-6 KO animals (Figure 3d). Injection of mouse recombinant IL-6 (1 μg/mouse, 4 hours) to IL-6 KO mice augmented ANXA1 expression after LPS injection, bringing it back to the degree of expression detected in WT mice (Figure 4). In this set of experiments an approximate eightfold increase in ANXA1 protein was measured in LPS-treated WT animals, and a value of 9 ± 0.5-fold increase was attained in IL-6 KO mice treated with LPS + IL-6. Interestingly, IL-6 alone was not sufficient to promote ANXA1 expression (Figure 4). Next, the effect of TNF-α, which levels varied between the two types of animals after LPS, was tested alone or together with IL-6. Injected at the dose of 1 μg per mouse, TNF-α significantly induced liver ANXA1 expression in WT animals, at levels similar to those achieved with LPS. Quite surprisingly, a pronounced response was also measured in IL-6 KO mice (Figure 4). However, co-injection with IL-6 (1 μg) produced maximal ANXA1 expression in the latter animals. To verify if IL-6 or TNF-α was able to target the ANXA1 expression directly on the hepatocyte, primary cultured hepatocytes prepared from perfused livers of WT or IL-6 KO animals were treated with the two different cytokines. In cells taken from WT mice, exogenous IL-6 induced an up-regulation of ANXA1 expression already at 3 hours (Figure 5A). In contrast IL-6 KO animal primary hepatocytes needed a prolonged (24 hours) incubation with IL-6 (10 ng/ml) to produce a significant response. A similar effect was seen with TNF-α (2 ng/ml), although the response to this cytokine was more rapid in primary hepatocytes from both WT and KO (Figure 5, A and B). There was no synergism or additive effect when the cytokines were co-incubated for 24 hours. Between 50 and 60% inhibition of LPS-induced liver ANXA1 protein expression was measured in WT and IL-6 KO mice pretreated with an anti-TNF-α antibody (Figure 6). In this set of experiments, the effect of the glucocorticoid antagonist RU-486 was also tested. RU-486 (20 mg/kg) was highly effective in WT mice (∼60% reduction in the ANXA1 response), whereas it was significantly less active in IL-6 KO mice (Figure 6). A mutual exclusion and certainly not an additive effect were observed when the two treatments were given together. The effect of an anti-IL-6 antibody was tested in WT mice: comparable inhibition (between 30 and 45%) in liver ANXA1 protein expression was measured, again without an additive effect with the glucocorticoid antagonist RU-486 (Figure 6). In IL-6 KO mice, TNF-α release was twofold to threefold higher than in WT mice (see Figure 2). As suggested by Fattori and colleagues,5Fattori E Cappelletti M Costa P Sellitto C Cantoni L Carelli M Faggioni R Fantuzzi G Ghezzi P Poli V Defective inflammatory response in interleukin 6-deficient mice.J Exp Med. 1994; 180: 1243-1250Crossref PubMed Scopus (487) Google Scholar this indicates that endogenous IL-6 controls TNF-α production during endotoxemia. Because ANXA1 is up-regulated by IL-6, and because IL-6 modulates TNF-α production, we next tested if ANXA1 participates in the modulation of TNF-α release. Figure 7 illustrates these data, with injection of mouse recombinant IL-6 to IL-6 KO mice being able to significantly reduce TNF-α (Figure 7A) but not IL-1 β (Figure 7B) plasma levels measured after LPS. We then tested the effect of recombinant ANXA1. In this series of experiments, LPS released >2 ng/ml of TNF-α as measured at 90 minutes after injection. A significant reduction (−25%) in TNF-α or (77%) IL-1 β was measured in the group of animals treated with ANXA1, whereas the structurally related protein ANXA5 was essentially inactive (Figure 7). This study stems from the investigation of Fattori and colleagues,5Fattori E Cappelletti M Costa P Sellitto C Cantoni L Carelli M Faggioni R Fantuzzi G Ghezzi P Poli V Defective inflammatory response in interleukin 6-deficient mice.J Exp Med. 1994; 180: 1243-1250Crossref PubMed Scopus (487) Google Scholar in which the liver response to systemic and local inflammation, produced with LPS and turpentine, respectively, was studied in the mouse. At variance from serum amyloid protein A,5Fattori E Cappelletti M Costa P Sellitto C Cantoni L Carelli M Faggioni R Fantuzzi G Ghezzi P Poli V Defective inflammatory response in interleukin 6-deficient mice.J Exp Med. 1994; 180: 1243-1250Crossref PubMed Scopus (487) Google Scholar ANXA1 expression in the liver was not modified by intramuscular injection of turpentine oil in either WT or IL-6 KO mice. In contrast, marked liver ANXA1 gene and protein synthesis were induced by LPS, and this was attenuated in IL-6 KO mice. We therefore conclude that liver ANXA1 is not overexpressed as a result of local inflammation (eg, as the one induced by turpentine) but rather as the result of a systemic inflammatory condition (eg, experimentally reproduced with LPS). For this reason, turpentine-induced inflammation was no longer used in the subsequent experiments. LPS injection to the experimental animal initiates a cascade of events that has been investigated in detail throughout the past two decades, characterized by a time-dependent and concerted release of proinflammatory cytokines.29Paludan SR Synergistic action of pro-inflammatory agents: cellular and molecular aspects.J Leukoc Biol. 2000; 67: 18-25Crossref PubMed Scopus (154) Google Scholar, 30Hersh D Weiss J Zychlinsky A How bacteria initiate inflammation: aspects of the emerging story.Curr Opin Microbiol. 1998; 1: 43-48Crossref PubMed Scopus (54) Google Scholar, 31Zetterstrom M Sundgren-Andersson AK Ostlund P Bartfai T Delineation of the proinflammatory cytokine cascade in fever induction.Ann NY Acad Sci. 1998; 856: 48-52Crossref PubMed Scopus (78) Google Scholar Monocyte/macrophage-derived TNF-α is the first cytokine release by LPS with plasma peak around 90 minutes,5Fattori E Cappelletti M Costa P Sellitto C Cantoni L Carelli M Faggioni R Fantuzzi G Ghezzi P Poli V Defective inflammatory response in interleukin 6-deficient mice.J Exp Med. 1994; 180: 1243-1250Crossref PubMed Scopus (487) Google Scholar, 15Zuckerman SH Shellhaas J Butler LD Differential regulation of lipopolysaccharide-induced interleukin 1 and tumor necrosis factor synthesis: effects of endogenous and exogenous glucocorticoids and the role of the pituitary-adrenal axis.Eur J Immunol. 1989; 19: 301-305Crossref PubMed Scopus (302) Google Scholar, 32Beutler B Krochin N Milsark IW Luedke C Cerami A Control of cachectin (tumor necrosis factor) synthesis: mechanisms of endotoxin resistance.Science. 1986; 232: 977-980Crossref PubMed Scopus (1009) Google Scholar and high levels of TNF-α were also measured in our experimental c" @default.
• W2039226089 created "2016-06-24" @default.
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• W2039226089 date "2001-10-01" @default.
• W2039226089 modified "2023-09-23" @default.
• W2039226089 title "Cytokine Modulation of Liver Annexin 1 Expression during Experimental Endotoxemia" @default.
• W2039226089 cites W1597308548 @default.
• W2039226089 cites W1613504607 @default.
• W2039226089 cites W1965231932 @default.
• W2039226089 cites W1965612828 @default.
• W2039226089 cites W1966324047 @default.
• W2039226089 cites W1967990939 @default.
• W2039226089 cites W1970814200 @default.
• W2039226089 cites W1974392294 @default.
• W2039226089 cites W1978067969 @default.
• W2039226089 cites W1980563273 @default.
• W2039226089 cites W1985018359 @default.
• W2039226089 cites W1986992784 @default.
• W2039226089 cites W1987150557 @default.
• W2039226089 cites W1992375878 @default.
• W2039226089 cites W2000294771 @default.
• W2039226089 cites W2004317005 @default.
• W2039226089 cites W2006392093 @default.
• W2039226089 cites W2006708761 @default.
• W2039226089 cites W2012410727 @default.
• W2039226089 cites W2018289835 @default.
• W2039226089 cites W2018870992 @default.
• W2039226089 cites W2019679024 @default.
• W2039226089 cites W2025126559 @default.
• W2039226089 cites W2028751330 @default.
• W2039226089 cites W2044648187 @default.
• W2039226089 cites W2052747333 @default.
• W2039226089 cites W2057018521 @default.
• W2039226089 cites W2058583204 @default.
• W2039226089 cites W2067624413 @default.
• W2039226089 cites W2067761476 @default.
• W2039226089 cites W2073015829 @default.
• W2039226089 cites W2073077060 @default.
• W2039226089 cites W2075787303 @default.
• W2039226089 cites W2077057589 @default.
• W2039226089 cites W2078690661 @default.
• W2039226089 cites W2095405203 @default.
• W2039226089 cites W2100837269 @default.
• W2039226089 cites W2100863754 @default.
• W2039226089 cites W2104083541 @default.
• W2039226089 cites W2109219945 @default.
• W2039226089 cites W2113828544 @default.
• W2039226089 cites W2116459589 @default.
• W2039226089 cites W2128214423 @default.
• W2039226089 cites W2131154808 @default.
• W2039226089 cites W2137472494 @default.
• W2039226089 cites W2138168133 @default.
• W2039226089 cites W2139416004 @default.
• W2039226089 cites W2145078103 @default.
• W2039226089 cites W2148285252 @default.
• W2039226089 cites W2149615892 @default.
• W2039226089 cites W2154746915 @default.
• W2039226089 cites W2162656941 @default.
• W2039226089 cites W2169563132 @default.
• W2039226089 cites W2200443534 @default.
• W2039226089 cites W2317184793 @default.
• W2039226089 cites W2395121051 @default.
• W2039226089 cites W2402677984 @default.
• W2039226089 cites W2415597251 @default.
• W2039226089 cites W308660587 @default.
• W2039226089 cites W4298193420 @default.
• W2039226089 cites W4300659364 @default.
• W2039226089 cites W78635780 @default.
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