ABSTRACT
Several lines of evidence indicate that 15-lipoxygenase type 1 (15-LO-1) plays a pathophysiological role in asthma. The aim for this study was to investigate the 15-LO-1 expression and activity in primary human airway epithelial cells cultivated on micro-porous filters at air-liquid interface. Incubation of human airway epithelial cells with arachidonic acid led to the formation of 15(S)-hydroxy-eicosatetraenoic acid (15-HETE) and exposing the cells to bacteria or physical injury markedly increased their production of 15-HETE. The cells were also found to convert arachidonic acid to eoxin C4 (EXC4). Subcellular fractionation revealed that the conversion of EXA4 to EXC4 was catalyzed by a soluble glutathione transferase (GST). The GST P1-1 enzyme was found to possess the highest activity of the investigated soluble GSTs. Following IL-4 treatment of airway epithelial cells, microarray analysis confirmed high expression of 15-LO-1 and GST P1-1, and immunohistochemical staining of bronchial biopsies revealed co-localization of 15-LO-1 and GST P1-1 in airway epithelial cells. These results indicate that respiratory infection and cell injury may activate the 15-LO pathway in airway epithelial cells. Furthermore, we also demonstrate that airway epithelial cells have the capacity to produce EXC4.
Subject(s)
Bronchi/cytology , Epithelial Cells/metabolism , Hydroxyeicosatetraenoic Acids/biosynthesis , Leukotrienes/biosynthesis , Arachidonate 15-Lipoxygenase/metabolism , Arachidonic Acid/metabolism , Biocatalysis , Cell Line , Glutathione Transferase/chemistry , Glutathione Transferase/metabolism , Humans , Protein Transport , SolubilityABSTRACT
BACKGROUND AND OBJECTIVE: Analysis of inflammatory biomarkers in saliva could offer an attractive opportunity for the diagnosis of different systemic conditions specifically in epidemiological surveys. The aim of this study was to investigate if certain salivary biomarkers could be used for detection of common systemic diseases. MATERIALS AND METHODS: A randomly selected sample of 1000 adults living in Skåne, a county in the southern part of Sweden, was invited to participate in a clinical study of oral health. 451 individuals were enrolled in this investigation, 51% women. All participants were asked to fill out a questionnaire, history was taken, a clinical examination was made and stimulated saliva samples were collected. Salivary concentrations of IL-1ß, -6, -8, TNF-α, lysozyme, MMP-8 and TIMP-1 were determined using ELISA, IFMA or Luminex assays. RESULTS: Salivary IL-8 concentration was found to be twice as high in subjects who had experience of tumour diseases. In addition, IL-8 levels were also elevated in patients with bowel disease. MMP-8 levels were elevated in saliva from patients after cardiac surgery or suffering from diabetes, and muscle and joint diseases. The levels of IL-1ß, IL-8 and MMP-8, as well as the MMP-8/TIMP-1 ratio were higher in subjects with muscle and joint diseases. CONCLUSION: Biomarkers in saliva have the potential to be used for screening purposes in epidemiological studies. The relatively unspecific inflammatory markers used in this study can not be used for diagnosis of specific diseases but can be seen as markers for increased systemic inflammation.
Subject(s)
Biomarkers/metabolism , Saliva/metabolism , Adult , Aged , Aged, 80 and over , Cytokines/metabolism , Epidemiologic Studies , Female , Humans , Inflammation Mediators/metabolism , Male , Middle Aged , Population Surveillance , Sweden/epidemiology , Young AdultABSTRACT
AIM: Saliva is a useful diagnostic fluid for oral-related diseases. Monitoring salivary biomarkers for oral and systemic diseases could become an important complement to clinical examinations in epidemiological surveys. Recent findings indicate that it is possible to detect biomarkers for oral diseases within saliva samples. The aim of this study was to investigate if known salivary biomarkers could be used for epidemiological studies for detection of periodontitis. MATERIALS AND METHODS: A randomly selected sample of adults (20-89 years) living in Southern Sweden were invited to participate. Four hundred and fifty-one individuals were examined clinically using standard examination procedures. Stimulated saliva samples were collected and analysed for concentrations of IL-1ß, -6, -8, lysozyme, matrix metalloproteinases (MMP)-8 and tissue inhibitor of metalloproteinase (TIMP)-1 using ELISA, immunofluorometric assay or Luminex assays. RESULTS: Patients with severe periodontitis presented with elevated salivary concentrations of IL-1ß (p < 0.001) and MMP-8 (p < 0.001). In addition, the MMP-8/TIMP-1 ratio was significantly higher in the severe periodontitis group (p < 0.001). Smokers compared with non-smokers showed slightly lower concentrations of IL-8 (p < 0.05) and MMP-8 (p = 0.052). CONCLUSION: This investigation shows that IL-1ß, MMP-8 and the ratio of MMP-8/TIMP-1 could be used as markers of periodontal disease in larger patient populations.
Subject(s)
Chronic Periodontitis/metabolism , Interleukin-1beta/analysis , Matrix Metalloproteinase 8/analysis , Saliva/chemistry , Salivary Proteins and Peptides/analysis , Tissue Inhibitor of Metalloproteinase-1/analysis , Adult , Aged , Aged, 80 and over , Analysis of Variance , Biomarkers , Case-Control Studies , Cross-Sectional Studies , Female , Humans , Linear Models , Male , Middle Aged , Oral Health , Periodontal Index , Smoking/metabolism , Statistics, Nonparametric , Surveys and Questionnaires , Young AdultABSTRACT
Human 15-lipoxygenase-1 (LO) possesses mainly 15-lipoxygenase activity whereas the animal ortholog 12/15-LO possesses mainly 12-lipoxygenase activity. These findings have raised the question if studies on animals can predict the function of 15-LO-1 in human. In this study we have characterized the arachidonic acid metabolites formed by porcine 12/15-LO. Mini pigs were infected with a parasite to increase the number of blood eosinophils, which highly express 12/15-LO. Isolated porcine polymorphonuclear leukocytes (PMNL) were incubated with arachidonic acid and the produced metabolites were analysed with HPLC and mass spectrometry (MS). The cells were found to produce 15-hydroxyeicosatetraenoic acid (HETE) and 12-HETE at a ratio of 1:5. Furthermore 8,15-dihydroxyeicosatetraenoic acids (DiHETEs) and 14,15-DiHETE were formed. Based on HPLC, UV-spectroscopy and MS analysis it was found that porcine PMNL also produced eoxin (EX) C4. These results demonstrate that although porcine 12/15-LO possesses primarily 12-lipoxygenase activity, the enzyme can catalyse the formation of EXC(4).
Subject(s)
Arachidonate 12-Lipoxygenase/metabolism , Arachidonate 15-Lipoxygenase/metabolism , Leukocytes/metabolism , Leukotrienes/biosynthesis , Animals , Arachidonic Acid/metabolism , Cells, Cultured , Chromatography, High Pressure Liquid , Mass Spectrometry , Neutrophils/metabolism , SwineABSTRACT
Classical Hodgkin lymphoma (cHL) is characterized histologically by a minority of malignant Hodgkin Reed-Sternberg cells surrounded by abundant inflammatory cells, generally believed to be of major importance in the pathophysiology of the disease. Here, we present data that link inflammatory cell-derived arachidonic acid metabolites, the cysteinyl leukotrienes (CysLT), to the pathogenesis of cHL. Two HL cell lines, L1236 and KMH2, were shown to express functional CysLT(1) receptors, responding with a robust calcium signal upon leukotriene (LT) D(4) challenge. LTD(4) stimulated protein release of tumor necrosis factor-alpha, interleukin-6 and -8 by L1236 cells and interleukin-8 by KMH2 cells. Importantly, all these LTD(4)-induced effects were blocked by the CysLT(1) receptor-specific antagonist zafirlukast. Immunohistochemical studies of cHL biopsies and microarray analysis of microdissected cells revealed that the CysLT(1) receptor is expressed also by primary Hodgkin Reed-Sternberg cells. As these cells are surrounded by CysLT-producing eosinophils, macrophages and mast cells, our results suggest the CysLTs as mediators in the pathogenesis of cHL, contributing to the aberrant cytokine network of this lymphoma.
