elafin and Inflammation

Elafin and its precursor trappin-2 are known for their contribution to the physiological mucosal shield against luminal microbes. Such a contribution seems to be particularly relevant in the gut, where the exposure of host tissues to heavy loads of microbes is constant and contributes to mucosa-associated pathologies. The expression of trappin-2/elafin has been shown to be differentially regulated in diseases associated with gut inflammation. Accumulating evidence has demonstrated the protective effects of trappin-2/elafin in gut intestinal disorders associated with acute or chronic inflammation, or with gluten sensitization disorders. The protective effects of trappin-2/elafin in the gut are discussed in terms of their pleiotropic modes of action: acting as protease inhibitors, transglutaminase substrates, antimicrobial peptides or as a regulator of pro-inflammatory transcription factors. Further, the question of the therapeutic potential of trappin-2/elafin delivery at the intestinal mucosa surface is raised. Whether trappin-2/elafin mucosal delivery should be considered to ensure intestinal tissue repair is also discussed.

Topics: Elafin; Humans; Inflammation; Intestinal Diseases; Protease Inhibitors

Secretory leukocyte protease inhibitor (SLPI), a serine protease inhibitor, which was most commonly examined in mucosal fluids such as saliva, is a versatile molecule and plays non-redundant roles. In addition to its anti-protease activity, SLPI has been shown to express anti-bacterial, anti-viral, anti-fungal, and anti-inflammatory properties as well as participating in innate and adaptive immune responses, most of which has been well documented. Recently, it is reported that SLPI is expressed in adipocytes and adipose tissue where it could play an important feedback role in the resolution of inflammation. Furthermore, circulating SLPI has been shown to correlate with progressive metabolic dysfunction. Moreover, adenoviral gene delivery of elafin and SLPI attenuates nuclear factor-κB-dependent inflammatory responses of human endothelial cells and macrophages to atherogenic stimuli. This review contributes to unraveling the protective role of SLPI in obesity-related atherosclerosis development, and the potential role in preventing arterial plaque rupture.

Topics: Adipocytes; Adipose Tissue; Anti-Inflammatory Agents; Antioxidants; Atherosclerosis; Elafin; Humans; Inflammation; Obesity; Plaque, Atherosclerotic; Secretory Leukocyte Peptidase Inhibitor

Trappin-2/Elafin is a potent serine protease inhibitor which prevents excessive damage under inflammatory status. This "alarm-antiprotease" is locally expressed by epithelial cells and immune cells such as macrophages and γδ T cells. It has also been proven to modulate a wide range of parameters that are critical for the inflammation process like modulating the NFκB pathway, cytokine secretion and cell recruitment. In addition, Trappin-2/Elafin was shown to possess anti-microbial properties against different classes of pathogens including viruses, fungi and bacteria. Studies also linked Trappin-2/Elafin to either susceptibility or protection against inflammatory disease and infections, even though the mechanisms remains poorly understood. This review will discuss some of the pleiotropic effects displayed by Trappin-2/Elafin, and the properties that could be used to prevent infection or to protect against inflammation.

Topics: Bacterial Infections; Elafin; Epithelial Cells; Humans; Immunity; Inflammation; Inflammation Mediators; Models, Immunological; Mycoses; T-Lymphocytes; Virus Diseases

WAP (whey acidic protein) is an important whey protein present in milk of mammals. This protein has characteristic domains, rich in cysteine residues, called 4-DSC (four-disulfide core domain). Other proteins, mainly present at mucosal surfaces, have been shown to also possess these characteristic WAP-4-DSC domains. The present review will focus on two WAP-4-DSC containing proteins, namely SLPI (secretory leucocyte protease inhibitor) and trappin-2/elafin. Although first described as antiproteases able to inhibit in particular host neutrophil proteases [NE (neutrophil elastase), cathepsin-G and proteinase-3] and as such, able to limit maladaptive tissue damage during inflammation, it has become apparent that these molecules have a variety of other functions (direct antimicrobial activity, bacterial opsonization, induction of adaptive immune responses, promotion of tissue repair, etc.). After providing information about the 'classical' antiproteasic role of these molecules, we will discuss the evidence pertaining to their pleiotropic functions in inflammation and immunity.

Topics: Animals; Elafin; Humans; Immunity, Mucosal; Inflammation; Milk Proteins; Protease Inhibitors; Protein Structure, Tertiary; Secretory Leukocyte Peptidase Inhibitor; Tissue Distribution

It is now clear that NSPs (neutrophil serine proteases), including elastase, Pr3 (proteinase 3) and CatG (cathepsin G) are major pathogenic determinants in chronic inflammatory disorders of the lungs. Two unglycosylated natural protease inhibitors, SLPI (secretory leucocyte protease inhibitor) and elafin, and its precursor trappin-2 that are found in the lungs, have therapeutic potential for reducing the protease-induced inflammatory response. This review examines the multifaceted roles of SLPI and elafin/trappin-2 in the context of their possible use as inhaled drugs for treating chronic lung diseases such as CF (cystic fibrosis) and COPD (chronic obstructive pulmonary disease).

Topics: Aerosols; Anti-Bacterial Agents; Antifungal Agents; Elafin; Humans; Inflammation; Lung Diseases; Proteinase Inhibitory Proteins, Secretory; Secretory Leukocyte Peptidase Inhibitor; Serine Proteases; Serine Proteinase Inhibitors; Transglutaminases

Elafin is an endogenous human protein composed of an N-terminal transglutaminase substrate motif and a C-terminal WAP (whey acidic protein)-domain with antiproteolytic properties. Elafin is expressed predominantly in epithelial tissue and potently inhibits the neutrophil-derived serine proteases elastase and proteinase-3 by a competitive tight-binding mechanism. Furthermore, it inhibits EVE (endogenous vascular elastase). Studies on several animal models show that antiprotease augmentation with human elafin is an effective strategy in the treatment of inflammatory vascular, systemic and pulmonary diseases and of inflammation triggered by reperfusion injury. This raises the possibility that elafin might be effective in the treatment of a variety of human inflammatory diseases. In a Phase I clinical trial, elafin was well tolerated. Phase II trials are underway to investigate the therapeutic effects of elafin on post-operative inflammation and the clinical consequences of major surgery. Of particular interest is the reduction of post-operative morbidity after oesophagus cancer surgery, coronary artery bypass surgery and kidney transplantation.

Topics: Animals; Clinical Trials as Topic; Elafin; Humans; Inflammation; Lung Diseases; Protease Inhibitors; Serine Endopeptidases; Serine Proteinase Inhibitors; Vascular Diseases

Diabetic retinopathy (DR) is a frequent microvascular complication of diabetes mellitus, and inflammatory pathways have been linked to its pathogenesis. In this retrospective, observational pilot study, we aimed to compare the concentrations of four inflammation-related proteins, ZAG, Reg-3a, elafin and RBP-4, in the serum and aqueous humor of healthy controls and diabetic patients with different stages of DR. The concentrations of VEGF-A, IL-8, IL-6 were determined in parallel as internal controls. In the serum, we did not find significant differences in the concentrations of target proteins. In the aqueous humor, higher levels of ZAG, RBP-4, Reg-3a and elafin were observed in advanced nonproliferative DR (NPDR)/ proliferative DR (PDR) compared to controls. The levels of ZAG and RBP-4 were also higher in advanced NPDR/PDR than in nonapparent DR. Normalization of target protein concentrations to the aqueous humor total protein demonstrates that a spill-over from serum due to breakage of the blood-retina barrier only partially accounts for increased inflammation related markers in later stages. In conclusion, we found elevated levels of Reg-3a, RBP-4, elafin and ZAG in advanced stages of diabetic retinopathy. Higher levels of pro-inflammatory proteins, Reg-3a and RBP-4, might contribute to the pathogenesis of diabetic retinopathy, as the parallel increased concentrations of anti-inflammatory molecules elafin and ZAG might indicate a compensatory mechanism.

Topics: Aqueous Humor; Diabetes Mellitus, Type 2; Diabetic Retinopathy; Elafin; Humans; Inflammation; Retrospective Studies

Topics: Arthritis; Autoimmunity; CCAAT-Enhancer-Binding Protein-beta; Cells, Cultured; Elafin; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Inflammation; Interleukin-4; Neutrophils; NF-kappa B; Osteoarthritis; Signal Transduction; Synovial Fluid; Tumor Necrosis Factor-alpha

Psoriasis is a pathological condition characterized by immune system dysfunction and inflammation. Patients with psoriasis are more likely to develop a wide range of disorders associated with inflammation. Serum levels of various substances and their combinations have been associated with the presence of the disease (psoriasis) and have shown the potential to reflect its activity. The aim of the present study is to contribute to the elucidation of pathophysiological links between psoriasis, its pro-inflammatory comorbidity metabolic syndrome (MetS), and the expression of clusterin and elafin, which are reflected in the pathophysiological "portfolio" of both diseases.. Clinical examinations (PASI score), ELISA (clusterin, elafin), and biochemical analyses (parameters of MetS) were performed.. We found that patients with psoriasis were more often afflicted by MetS, compared to the healthy controls. Clusterin and elafin levels were higher in the patients than in the controls but did not correlate to the severity of psoriasis.. Our data suggest that patients with psoriasis are more susceptible to developing other systemic inflammatory diseases, such as MetS. The levels of clusterin and elafin, which are tightly linked to inflammation, were significantly increased in the patients, compared to the controls, but the presence of MetS in patients did not further increase these levels.

Topics: Adult; Body Mass Index; Case-Control Studies; Clusterin; Comorbidity; Elafin; Female; Gene Expression Regulation; Humans; Inflammation; Male; Metabolic Syndrome; Middle Aged; Psoriasis; Severity of Illness Index

Receptor activator of NF-κB ligand (RANKL) as an osteoclast differentiation factor induces inflammatory reactions via production of thymic stromal lymphopoietin (TSLP). Epigallocatechin gallate (EGCG) is the major and the most active compound in green tea and has anti-inflammatory, anti-cancer, anti-oxidant, and neuroprotective effects. However, the effect and molecular mechanisms of EGCG are still unknown in RANKL-induced inflammatory reactions. Here we investigated the immuno-regulatory effects and its molecular mechanisms of epigallocatechin gallate (EGCG) in RANKL-stimulated human mast cell line, HMC-1 cells. In this study, EGCG prevented expression of PI3 Kinase and phosphorylation of mitogen-activated protein (MAP) Kinases in RANKL-stimulated HMC-1 cells. EGCG prevented caspase-1 activity and decreased transcriptional activity of nuclear factor (NF)-κB by suppressing inhibitory protein κBα phosphorylation in RANKL-stimulated HMC-1 cells. EGCG has been shown to prevent production and mRNA expression of TSLP, interleukin (IL)-1β, IL-6, and IL-8 by RANKL without cytotoxicity. Furthermore, EGCG prevented degranulation of mast cell in RANKL-stimulated HMC-1 cells. Overall, these results suggest that EGCG acts as a natural agent for preventing and treating RANKL-mediated inflammatory diseases by targeting PI3 Kinase, MAP Kinase, caspase-1, and NF-κB signaling cascade in mast cells.

Topics: Caspase 1; Catechin; Cell Line; Cell Survival; Cytokines; Elafin; Histamine; Humans; Inflammation; Interleukin-1beta; Interleukin-6; Interleukin-8; Mast Cells; Mitogen-Activated Protein Kinases; NF-kappa B; RANK Ligand; Signal Transduction; Thymic Stromal Lymphopoietin

Psoriasis is a distressing chronic skin disease. Its exact pathogenesis is still unclear, as many factors interplay in its occurrence.. The aim of the study is to estimate elafin levels in the serum of both cases and controls and to correlate the levels with psoriasis severity and other markers of inflammation.. Twenty-six psoriatic cases along with 26 healthy controls were assigned in this case-control study. Psoriasis severity was determined by Psoriasis Area and Severity Index score. Elafin levels were measured by ELISA in serum of both cases and controls. C-reactive protein (CRP) and erythrocyte sedimentation rate (ESR) were measured in the cases.. Elafin levels show highly statistical significance difference (P < 0.001) between cases and controls. There is a statistical significant correlation between elafin levels and both psoriasis severity and inflammation markers as CRP and ESR.. Elafin represents a mirror for psoriasis severity and inflammatory state.

Topics: Adolescent; Adult; Aged; Biomarkers; Blood Sedimentation; C-Reactive Protein; Case-Control Studies; Child; Elafin; Female; Humans; Inflammation; Male; Middle Aged; Psoriasis; Severity of Illness Index; Young Adult

Despite increasing evidence that adults with long-standing atopic dermatitis (AD) have systemic inflammation, little is known about systemic inflammation in recent-onset early pediatric AD.. To analyze blood inflammatory proteins of early pediatric AD.. Using high-throughput proteomics (proximity extension assay), we assessed 257 inflammatory and cardiovascular risk proteins in the blood of 30 children with moderate to severe AD younger than 5 years of age (within 6 months of onset) compared with age-matched pediatric control individuals and adult patients with AD.. In pediatric AD blood, T helper (Th) type 2 (CCL13, CCL22) and Th17 (peptidase inhibitor-3/elafin) markers were increased, together with markers of tissue remodeling (matrix metalloproteinases 3/9/10, urokinase receptor), endothelial activation (E-selectin), T-cell activation (IL2RA), neutrophil activation (myeloperoxidase), lipid metabolism (FABP4), and growth factors (FGF21, transforming growth factor-α). Total numbers of dysregulated proteins were smaller in pediatric AD (n = 22) than in adult AD (n = 61). Clinical severity scores were positively correlated with receptors for interleukins 33 and 36 and inversely correlated with some Th1 markers (interferon gamma, CXCL11).. Different baseline expression levels in healthy pediatric vs adult samples.. Within months of pediatric AD onset, systemic immune activation is present, with Th2/Th17 skewing but otherwise different proteomic patterns from adult AD. Future correlation of proteomic patterns with disease course, comorbidity development, and drug response may yield predictive biomarkers.

Topics: Age Factors; Biomarkers; Case-Control Studies; Chemokines; Child, Preschool; Chronic Disease; Dermatitis, Atopic; E-Selectin; Elafin; Fatty Acid-Binding Proteins; Female; Fibroblast Growth Factors; Humans; Infant; Inflammation; Interleukin-2 Receptor alpha Subunit; Male; Matrix Metalloproteinases; Peroxidase; Proteome; Receptors, Interleukin; RNA, Messenger; Severity of Illness Index; Transforming Growth Factor alpha

Elafin is a low molecular weight protein with antileukoproteinase, anti-inflammatory, antibacterial and immunomodulating properties. The profile of Elafin in fetal membranes is not well characterized. This study determined the changes in Elafin expression and concentration in human fetal membrane from patients with preterm prelabor rupture of membranes (PPROM) and in vitro in response to intra-amniotic polymicrobial pathogens.. Elafin messenger RNA (mRNA) expressions were studied in fetal membranes from PPROM, normal term as well as in normal term not in labor membranes in an organ explant system treated (24 h) with lipopolysaccharide (LPS), using quantitative reverse transcription-polymerase chain reaction (RT-PCR). Enzyme-linked immunosorbent assay (ELISA) measured Elafin concentrations in culture supernatants from tissues treated with LPS and polybacterial combinations of heat-inactivated Mycoplasma hominis (MH), Ureaplasma urealyticum (UU) and Gardnerella vaginalis (GV).. Elafin mRNA expression in fetal membranes from women with PPROM was significantly higher compared to women who delivered at term after normal pregnancy (5.09±3.50 vs. 11.71±2.21; P<0.05). In vitro, LPS-stimulated membranes showed a significantly increased Elafin m-RNA expression (P<0.05). However, the protein levels after LPS stimulation was not changed. Similarly, polymicrobial-treated fetal membranes also showed no changes in Elafin protein concentrations compared to untreated controls.. Higher Elafin expression in PPROM fetal membranes suggests a host response to an inflammatory pathology. However, lack of Elafin response to LPS and polymicrobial treatment is indicative of the minimal anti-inflammatory impact of this molecule in fetal membranes.

Topics: Elafin; Extraembryonic Membranes; Female; Fetal Membranes, Premature Rupture; Host-Pathogen Interactions; Humans; In Vitro Techniques; Inflammation; Lipopolysaccharides; Organ Culture Techniques; Pregnancy

Protease-activated receptor 2 (PAR2) is implicated in the pathogenesis of chronic inflammatory diseases, including periodontitis; it can be activated by gingipain and produced by Porphyromonas gingivalis and by neutrophil protease 3 (P3). PAR2 activation plays a relevant role in inflammatory processes by inducing the release of important inflammatory mediators associated with periodontal breakdown. The effects of periodontal treatment on PAR2 expression and its association with levels of proinflammatory mediators and activating proteases were investigated in chronic periodontitis patients. Positive staining for PAR2 was observed in gingival crevicular fluid cells and was reflective of tissue destruction. Overexpression of PAR2 was positively associated with inflammatory clinical parameters and with the levels of interleukin-6 (IL-6), IL-8, tumor necrosis factor alpha, matrix metalloprotease 2 (MMP-2), MMP-8, hepatocyte growth factor, and vascular endothelial growth factor. Elevated levels of gingipain and P3 and decreased levels of dentilisin and the protease inhibitors secretory leukocyte protease inhibitor and elafin were also associated with PAR2 overexpression. Healthy periodontal sites from individuals with chronic periodontitis showed diminished expression of PAR2 mRNA and the PAR2 protein (P < 0.05). Furthermore, periodontal treatment resulted in decreased PAR2 expression and correlated with decreased expression of inflammatory mediators and activating proteases. We concluded that periodontal treatment resulted in decreased levels of proteases and that proinflammatory mediators are associated with decreased PAR2 expression, suggesting that PAR2 expression is influenced by the presence of periodontal infection and is not a constitutive characteristic favoring periodontal inflammation.

Topics: Adhesins, Bacterial; Adult; Bacterial Proteins; Chymotrypsin; Cysteine Endopeptidases; Elafin; Female; Gingipain Cysteine Endopeptidases; Gingival Crevicular Fluid; Hepatocyte Growth Factor; Humans; Inflammation; Inflammation Mediators; Interleukin-6; Interleukin-8; Male; Matrix Metalloproteinase 2; Matrix Metalloproteinase 8; Middle Aged; Myeloblastin; Peptide Hydrolases; Periodontal Pocket; Periodontitis; Porphyromonas gingivalis; Receptor, PAR-2; RNA, Messenger; Secretory Leukocyte Peptidase Inhibitor; Tumor Necrosis Factor-alpha; Vascular Endothelial Growth Factor A; Young Adult

The etiology of inflammatory myofibroblastic tumors (IMTs) is controversial and the prognosis is unpredictable. Previous studies have not investigated the expression of hypoxia-related markers in IMTs.. Between 2002 and 2012, 12 consecutive patients with histologically proven IMTs were enrolled in the study. Immunohistochemistry was used to detect GLUT-1, HIF-1α, PI3K, and p-Akt expression in paraffin-embedded tumor specimens. Associations among GLUT-1, HIF-1α, PI3K, and p-Akt protein expression and clinical parameters were investigated.. The mean duration of follow-up was 52.1 months (range, 11 to 132 months). Six patients had local recurrence. GLUT-1, HIF-1α, PI3K, and p-Akt expression were detected in 41.7%, 50.0%, 33.3%, and 41.7% of patients, respectively. Fisher's exact test revealed significant correlations between recurrence of IMT and PI3K expression (P = 0.01) and p-Akt expression (P = 0.015). Univariate analyses revealed significant correlations between survival and GLUT-1 expression (P = 0.028), PI3K expression (P = 0.006), and p-Akt expression (P = 0.028). Multivariate analysis did not show a significant relationship between survival and GLUT-1, HIF-1α, PI3K, or p-Akt. Spearman rank correlation analysis showed significant correlations between HIF-1α and PI3K expression (r = 0.707, P = 0.01) and between p-Akt and PI3K expression (r = 0.837, P = 0.001).. Although our results are inconclusive owing to the small sample size, they suggest that PI3K and p-Akt expression may play a role in the recurrence of IMTs of the head and neck.

Topics: Adult; Biomarkers, Tumor; Elafin; Female; Follow-Up Studies; Glucose Transporter Type 1; Head and Neck Neoplasms; Humans; Hypoxia; Hypoxia-Inducible Factor 1, alpha Subunit; Immunoenzyme Techniques; Inflammation; Lymphatic Metastasis; Male; Middle Aged; Myofibroblasts; Neoplasm Recurrence, Local; Neoplasm Staging; Neoplasms, Muscle Tissue; Phosphatidylinositol 3-Kinases; Prognosis; Proto-Oncogene Proteins c-akt; Young Adult

Humanized mouse models offer a challenging possibility to study human cell function in vivo. In the huPBL-SCID-huSkin allograft model human skin is transplanted onto immunodeficient mice and allowed to heal. Thereafter allogeneic human peripheral blood mononuclear cells are infused intra peritoneally to induce T cell mediated inflammation and microvessel destruction of the human skin. This model has great potential for in vivo study of human immune cells in (skin) inflammatory processes and for preclinical screening of systemically administered immunomodulating agents. Here we studied the inflammatory skin response of human keratinocytes and human T cells and the concomitant systemic human T cell response.As new findings in the inflamed human skin of the huPBL-SCID-huSkin model we here identified: 1. Parameters of dermal pathology that enable precise quantification of the local skin inflammatory response exemplified by acanthosis, increased expression of human β-defensin-2, Elafin, K16, Ki67 and reduced expression of K10 by microscopy and immunohistochemistry. 2. Induction of human cytokines and chemokines using quantitative real-time PCR. 3. Influx of inflammation associated IL-17A-producing human CD4+ and CD8+ T cells as well as immunoregulatory CD4+Foxp3+ cells using immunohistochemistry and -fluorescence, suggesting that active immune regulation is taking place locally in the inflamed skin. 4. Systemic responses that revealed activated and proliferating human CD4+ and CD8+ T cells that acquired homing marker expression of CD62L and CLA. Finally, we demonstrated the value of the newly identified parameters by showing significant changes upon systemic treatment with the T cell inhibitory agents cyclosporine-A and rapamycin. In summary, here we equipped the huPBL-SCID-huSkin humanized mouse model with relevant tools not only to quantify the inflammatory dermal response, but also to monitor the peripheral immune status. This combined approach will gain our understanding of the dermal immunopathology in humans and benefit the development of novel therapeutics for controlling inflammatory skin diseases.

Topics: Animals; Antigens, Differentiation, T-Lymphocyte; beta-Defensins; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Cell Differentiation; Cyclosporine; Disease Models, Animal; Elafin; Gene Expression Regulation; Humans; Inflammation; Injections, Intraperitoneal; Interleukin-17; Keratinocytes; Keratins; Ki-67 Antigen; L-Selectin; Membrane Glycoproteins; Mice; Mice, SCID; Sirolimus; Skin; Skin Transplantation; Transplantation, Heterologous

Elafin, a natural protease inhibitor expressed in healthy intestinal mucosa, has pleiotropic anti-inflammatory properties in vitro and in animal models. We found that mucosal expression of Elafin is diminished in patients with inflammatory bowel disease (IBD). This defect is associated with increased elastolytic activity (elastase-like proteolysis) in colon tissue. We engineered two food-grade strains of lactic acid bacteria (LAB) to express and deliver Elafin to the site of inflammation in the colon to assess the potential therapeutic benefits of the Elafin-expressing LAB. In mouse models of acute and chronic colitis, oral administration of Elafin-expressing LAB decreased elastolytic activity and inflammation and restored intestinal homeostasis. Furthermore, when cultures of human intestinal epithelial cells were treated with LAB secreting Elafin, the inflamed epithelium was protected from increased intestinal permeability and from the release of cytokines and chemokines, both of which are characteristic of intestinal dysfunction associated with IBD. Together, these results suggest that oral delivery of LAB secreting Elafin may be useful for treating IBD in humans.

Topics: Animals; Bacteria; Colon; Elafin; Humans; In Vitro Techniques; Inflammation; Inflammatory Bowel Diseases; Intestinal Mucosa; Mice

To explore the effects of recombinant human Elafin gene transfection on lipopolysaccharide (LPS)-induced airway mucous hypersecretion.. An eukaryotic expression vector pEGFP-C1-Elafin was first constructed and then transfected into NCI-H292 cell. After co-incubation with polymorphonuclear granulocyte (PMN) plus LPS stimulation for 24 hour, the vital force unit of neutrophil elastase (NE) and the content of mucin (MUC) protein in culture media and the level of MUC 5AC mRNA in culture cells were detected with substrate detection technique, enzyme-linked immunosorbent assay (ELISA) and reverse transcription-polymerase chain reaction (RT-PCR) respectively.. All parameters in control group showed a low level (0.13 ± 0.03, 0.28 ± 0.06, 0.29 ± 0.04 respectively). And transfecting NCI-H292 cell with pEGFP-C1 at different levels of LPS, all parameters were obviously different (all P < 0.01) and they all showed a dose-dependent relationship. After LPS stimulation (0.5, 5 mg/L), as compared with the transfection of NCI-H292 cell with pEGFP-C1-Elafin, the vital force unit of NE and the content of MUC5AC protein in culture media and the level of MUC 5AC mRNA in transfecting pEGFP-C1 NCI-H292 cell decreased obviously (all P < 0.05). But after LPS stimulation (25 mg/L), as compared with the transfection of NCI-H292 cell with pEGFP-C1-Elafin, no parameter of transfecting pEGFP-C1 NCI-H292 cell had any obvious alteration (0.67 ± 0.06, 0.64 ± 0.08, 0.67 ± 0.07 respectively, all P > 0.05).. The transfection of recombinant human elafin gene into NCI-H292 cell may inhibit the force of inflammatory telo-effective factor NE induced by a certain level of LPS and airway mucous hypersecretion. But it can not inhibit the effects induced by a high level of LPS.

Topics: Cell Line; Elafin; Genetic Vectors; Humans; Inflammation; Leukocyte Elastase; Lipopolysaccharides; Mucin 5AC; Mucins; Recombinant Proteins; Respiratory Mucosa; Transcription, Genetic; Transfection

Elafin, an antiprotease, is likely to protect pulmonary epithelial cells from inflammatory damages. We aimed to explore the molecule mechanisms of recombinant human elafin on protecting A549 cells integrity against inflammatory assault. We transfected A549 airway epithelial cells with eukaryotic expression vector pEGFP-N1-Elafin and negative control vector pEGFP-N1, respectively. Cells were co-incubated with polymorphonuclear neutrophils (PMN) and then were stimulated with lipopolysaccharide (LPS). Results revealed that, in pEGFP-N1-Elafin transfected cells, neutrophil elastase (NE) activity significantly decreased after LPS stimulation, accompanied with elevated elafin mRNA level and protein production, whereas in cells transfected with pEGFP-N1, NE activity was higher and elafin expression was lower, compared with pEGFP-N1-Elafin transfected group (P < 0.05). In pEGFP-N1 transfected cells, LPS suppressed tight junctions protein zonula occludens-1 (ZO-1) production, while in recombinant pEGFP-N1-Elafin cells, LPS did not cause significantly decrease of ZO-1, compared with normal control cells. LPS stimulation also significantly weakened the collagen adhesion capability of pEGFP-N1 transfected cells (P < 0.01), but there was no significant difference in recombinant pEGFP-N1-Elafin cells (P > 0.05). These results suggest that elafin can maintain airway epithelium integrity by protecting airway epithelial cells and enhancing the anti-inflammatory capability of airway.

Topics: Aged; Cell Adhesion; Cell Line, Tumor; Collagen; Elafin; Fluorescent Antibody Technique; Gene Expression Regulation, Neoplastic; Humans; Inflammation; Leukocyte Elastase; Lipopolysaccharides; Male; Membrane Proteins; Microscopy, Confocal; Middle Aged; Phosphoproteins; Recombinant Proteins; Respiratory Mucosa; RNA, Messenger; Subcellular Fractions; Zonula Occludens-1 Protein

Elafin (or skin-derived antileukoprotease) and secretory leukocyte protease inhibitor (SLPI) are serine antiproteases antagonizing human neutrophil elastase (HNE), thereby preventing tissue injury from excessive release of proteolytic enzymes by inflammatory cells. Furthermore, elafin and SLPI are "defensin-like" molecules with broad antimicrobial activity. The balance between proteases and antagonists may critically determine inflammatory processes in Crohn's disease (CD) and ulcerative colitis (UC). Real-time PCR was performed to quantitate colonic, proinflammatory cytokine IL-8, protease (HNE), and antiprotease mRNA (elafin and SLPI) in a total of 340 biopsies from 117 patients (47 CD, 45 UC, 25 controls). Histological inflammation was scored, and HNE, elafin, and SLPI were localized and semiquantified by immunostaining in 51 colonic paraffin sections (23 CD, 11 UC, 17 controls). Proinflammatory IL-8, degree of histological inflammation, and granulocyte content were similar in UC and CD. Elafin stained predominantly in the epithelium and SLPI in mucosal inflammatory cells. HNE mRNA levels and immunostaining were increased equally in both forms of inflammatory bowel disease. Levels of mRNA and immunostaining of the antiproteases elafin and SLPI were enhanced strongly in inflamed versus noninflamed UC. It is surprising that comparing inflamed versus noninflamed CD, this increase was significantly less pronounced for elafin and even lacking for SLPI. Despite comparable degrees of inflammation and protease levels, the induction of both antiproteases was attenuated in CD. This could contribute to the transmural depth of tissue destruction in CD. Elafin and SLPI may be added to the list of defensin-like peptides with diminished induction in CD versus UC.

Topics: Adult; Case-Control Studies; Colon; Crohn Disease; Elafin; Epithelial Cells; Frameshift Mutation; Humans; Inflammation; Intestinal Mucosa; Leukocyte Elastase; Leukocytes; Middle Aged; Nod2 Signaling Adaptor Protein; Secretory Leukocyte Peptidase Inhibitor