leukotriene-e4 and Inflammation

The review discusses what is known regarding airway molecular phenotypes in pediatric asthma, specifically biomarkers that have been studied and their relation to the various clinical phenotypes of asthma.. Pediatric asthma is a complex and heterogeneous disease that consists of several clinical phenotypes. There have been numerous studies investigating inflammatory markers that would increase our understanding of the underlying pathogenesis of asthma as well as facilitate the discovery of therapies for these patients. Some of these biomarkers, such as exhaled nitric oxide, exhaled breath condensate, urine leukotriene E4 and induced sputum are less invasive measures of inflammation than obtaining bronchoalveolar lavage fluid in children. Although recent data reveal that some of these measures may be helpful in classifying and managing pediatric asthma, further studies are critically needed before any of these biomarkers are able to be routinely used in clinical asthma care.. The search for noninvasive biomarkers to help elucidate specific underlying molecular phenotypes in pediatric asthma should be a continued priority as we work towards improved care and management of these children.

Topics: Adolescent; Asthma; Biomarkers; Child; Child, Preschool; Humans; Inflammation; Inflammation Mediators; Leukotriene E4; Lung; Nitric Oxide; Phenotype; Sputum

Cysteinyl leukotrienes (CysLTs: leukotrienes C(4), D(4), and E(4)) have long been implicated in the pathogenesis of asthma and several allergic diseases. LTE(4) has been identified as a major metabolite of LTC(4), and urinary LTE(4) (U-LTE(4)) is considered as the most reliable analytic parameter for monitoring the endogenous synthesis of CysLTs. From recent studies on the U-LTE(4) associated with adult stable asthma we identified four factors for hyperleukotrieneuria, namely, aspirin intolerance, eosinophilic nasal polyposis (ENP), vasculitis, and severe asthma. In ENP, there is prominent infiltration of eosinophils in the sinus and polyp tissues, which is linked to adult asthma and aspirin sensitivity, and ENP is the most important factor for the overproduction of CysLTs in asthmatics. We also demonstrated that anaphylaxis and eosinophilic pneumonia (EP) are associated with a marked increase in the U-LTE(4) concentration. Under these disease conditions, U-LTE(4) may be one of the candidate biomarkers. Moreover, the changes in U-LTE(4) concentrations may provide valuable information concerning therapeutic targets.

Topics: Aspirin; Asthma; Biomarkers; Cell Movement; Drug Hypersensitivity; Eosinophils; Gene Expression Regulation; Inflammation; Leukotriene E4; Nasal Polyps; Risk Factors; Vasculitis

Topics: Asthma; Basophils; Cell Communication; Endothelium, Vascular; Eosinophils; Humans; Inflammation; Leukotriene B4; Leukotriene E4; Mast Cells; SRS-A

Topics: Animals; Cardiovascular System; Coronary Disease; Dogs; Guinea Pigs; Hemodynamics; Histamine; Humans; Inflammation; Leukotriene E4; Macaca; Rabbits; SRS-A

Immunological and non-immunological injury induce as a result of the action of the enzyme lipoxygenase the release of a series of arachidonic acid metabolites known as leukotrienes. The leukotrienes play an important role in allergic and inflammatory disease. Leukotrienes C4, D4 and E4 which recently have been recognized as constituents of the allergic mediator slow reacting substance of anaphylaxis (SRS-A) induce powerful bronchoconstriction, plasma exudation and weal and flare responses. Leukotriene B4 is involved in the regulation of chemotaxis, chemokinesis and other aspects of both cellular and vascular inflammation. The development of specific lipoxygenase inhibitors may lead to a new class of drugs for the treatment of bronchial asthma and chronic inflammatory diseases.

Topics: Arachidonic Acids; Asthma; Biotransformation; Chemotaxis, Leukocyte; Humans; Hypersensitivity; Inflammation; Leukotriene A4; Leukotriene B4; Leukotriene E4; Lipoxygenase; Lipoxygenase Inhibitors; Neutrophils; SRS-A

In this study, prospectively, we aimed to determine the effects of the different treatment alternatives on the oxidant system and inflammatory and clinic determinants during the stable period of 1 month following an asthmatic attack. Thirty-one patients (22 female, nine male) were randomly divided into three groups following the stabilization of an acute asthma attack. The control group that is an additional group to the three patient groups consisted of 10 healthy volunteers (five female, five male). The following protocols were used for 4 weeks: Group I: short-acting inhaler beta2 mimetic as required (treatment A)+800 mug inhaler budesonide (treatment B)+leukotriene receptor antagonist; Group II: treatment A and B; Group III: treatment A and B+vitamin E. The serum levels before and after treatment of eosinophilic cationic protein (ECP), leukotriene E4 (LTE(4)), and malondialdehyde (MDA) were determined. The values before and after treatment were statistically compared both with each other and control values. Pretreatment ECP, LTE(4), and MDA levels for the three groups were significantly higher compared with post-treatment levels (P<0.05 to P<0.001) and the control levels (P<0.01 to P<0.001). However, when post-treatment levels were compared with those of the control group, no significant differences were found (P>0.05). Lack of significant variation was observed when the pre- and post-treatment differences in the three groups were compared for each one of ECP, LTE(4), and MDA levels (P>0.05). Leukotriene receptor antagonist or antioxidant agents added to standard asthma treatment did not make a significant contribution on ECP, LTE(4), and MDA levels and respiratory parameters such as spirometric function tests. Etiologic factors and/or the possible changes in different pathogenetic ways of the inflammation process may have been responsible for nonsignificant intertreatment difference in the biomarker levels. The result confirms that suppressing the inflammation in asthma enables the entire inflammatory pathologic process to be controlled.

Topics: Adult; Anti-Asthmatic Agents; Antioxidants; Asthma; Biomarkers; Female; Forced Expiratory Volume; Humans; Immunoenzyme Techniques; Inflammation; Leukotriene Antagonists; Leukotriene E4; Male; Malondialdehyde; Oxidative Stress; Oxygen; Respiratory Mechanics; Spirometry

Bronchoalveolar lavage (BAL) was used to sample lung cells and biochemical components in the lung air spaces at various times from 1 to 91 d after intrapulmonary instillation of 2.6 microm-diameter iron oxide particles in human subjects. The instillation of particles induced transient acute inflammation during the first day post instillation (PI), characterized by increased numbers of neutrophils and alveolar macrophages as well as increased amounts of protein, lactate dehydrogenase, and interleukin-8 in BAL fluids. This response was subclinical and was resolved within 4 d PI. A similar dose-dependent response was seen in rats 1 d after intratracheal instillation of the same particles. The particles contained small amounts of soluble iron (240 ng/mg) and possessed the capacity to catalyze oxidant generation in vitro. Our findings indicate that the acute inflammation after particle exposure may, at least partially, be the result of oxidant generation catalyzed by the presence of residual amounts of ferric ion, ferric hydroxides, or oxyhydroxides associated with the particles. These findings may have relevance to the acute health effects associated with increased levels of ambient particulate air pollutants.

Topics: Adult; Animals; Bronchoalveolar Lavage; Bronchoalveolar Lavage Fluid; Dinoprostone; Female; Ferric Compounds; Humans; Inflammation; Instillation, Drug; Interleukin-8; Iron; L-Lactate Dehydrogenase; Leukotriene C4; Leukotriene E4; Lung; Macrophages, Alveolar; Male; Neutrophils; Phagocytosis; Rats; Time Factors

To investigate the contribution of leukotrienes (LTs) to inflammation and bronchoconstriction in nocturnal asthma, we performed a randomized trial in 12 asthmatic patients and 6 normal control subjects. This study involved pulmonary function testing, methacholine challenge, bronchoscopy for cell counts, LT and thromboxane (TX) levels in bronchoalveolar lavage (BAL) fluid, and collection of urine for LTs at 4:00 P.M. and 4:00 A.M. At 4:00 P.M. BAL fluid LTB4 and sulfidopeptide LT levels in asthmatic and control subjects were not statistically different. At 4:00 A.M. alone, LTB4 and cysteinyl LT levels increased to become significantly greater in asthmatic than in control subjects, LTB4 levels correlating significantly (r = -0.66, p < 0.0001) with nocturnal fall in FEV1. Nocturnal asthmatic urinary LTE4 levels were also significantly higher than those of control subjects. The 4:00 A.M. testing was repeated during treatment with a 5-lipoxygenase inhibitor, zileuton. In asthmatic subjects, zileuton decreased BAL fluid LTB4 (p = 0.01) and urinary LTE4 (p = 0.01) while showing a trend for improving nocturnal FEV1 (p = 0.086). These decreases in LTB4 levels and improvement in FVE1 were associated with significant reductions in 4 A.M. BAL fluid and blood eosinophil percentages on zileuton compared with placebo administration. These findings demonstrate the importance of LTs in both the inflammation and the physiology of nocturnal asthma.

Topics: Adolescent; Adult; Asthma; Bronchial Provocation Tests; Bronchoalveolar Lavage Fluid; Bronchoconstriction; Cross-Over Studies; Double-Blind Method; Forced Expiratory Volume; Humans; Hydroxyurea; Inflammation; Leukotriene E4; Leukotrienes; Lipoxygenase Inhibitors; Middle Aged

Topics: Adult; Asthma; Biomarkers; Female; Humans; Inflammation; Leukotriene E4; Male; Middle Aged; Prostaglandins

Topics: Adult; Asthma; Child; Diabetes Mellitus, Type 2; Humans; Inflammation; Leukotriene E4; Prostaglandin D2

Nonsteroidal anti-inflammatory drug (NSAID)-exacerbated respiratory disease (N-ERD) asthma is characterized by chronic rhinosinusitis and intolerance of aspirin and other COX1 inhibitors. Clinical data point to a heterogeneity within the N-ERD phenotype.. Our aim was to investigate immune mediator profiles in the lower airways of patients with N-ERD.. Levels of cytokines (determined by using Luminex assay) and eicosanoids (determined by using mass spectrometry) were measured in bronchoalveolar lavage fluid (BALF) from patients with N-ERD (n = 22), patients with NSAID-tolerant asthma (n = 21), and control subjects (n = 11). mRNA expression in BALF cells was quantified by using TaqMan low-density arrays.. Lower airway eosinophilia was more frequent in N-ERD (54.5%) than in NSAID-tolerant asthma (9.5% [P = .009]). The type-2 (T2) immune signature of BALF cells was more pronounced in the eosinophilic subphenotype of N-ERD. Similarly, BALF concentrations of periostin and CCL26 were significantly increased in eosinophilic N-ERD and correlated with T2 signature in BALF cells. Multiparameter analysis of BALF mediators of all patients with asthma revealed the presence of 2 immune endotypes: T2-like (with an elevated level of periostin in BALF) and non-T2/proinflammatory (with higher levels of matrix metalloproteinases and inflammatory cytokines). Patients with N-ERD were classified mostly as having the T2 endotype (68%). Changes in eicosanoid profile (eg, increased leukotriene E. Lower airway immune profiles show considerable heterogeneity of N-ERD, with skewing toward T2 response and eosinophilic inflammation. Increased production of leukotriene E

Topics: Adult; Aged; Anti-Inflammatory Agents, Non-Steroidal; Aspirin; Asthma; Biomarkers; Bronchoalveolar Lavage Fluid; Eosinophilia; Eosinophils; Female; Humans; Inflammation; Leukotriene E4; Male; Middle Aged; Nasal Lavage; Neutrophils; Rhinitis; Sinusitis

Cysteinyl leukotriene (cysLT) overproduction and eosinophil activation are hallmarks of aspirin-exacerbated respiratory disease (AERD). However, pathogenic mechanisms of AERD remain to be clarified. Here, we aimed to find the significance of transforming growth factor beta 1 (TGF-β1) in association with cysteinyl leukotriene E4 (LTE4) production, leading to eosinophil degranulation. To evaluate levels of serum TGF-β1, first cohort enrolled AERD (n = 336), ATA (n = 442) patients and healthy control subjects (HCs, n = 253). In addition, second cohort recruited AERD (n = 34) and ATA (n = 25) patients to investigate a relation between levels of serum TGF-β1 and urinary LTE4. The function of TGF-β1 in LTE4 production was further demonstrated by ex vivo (human peripheral eosinophils) or in vivo (BALB/c mice) experiment. As a result, the levels of serum TGF-β1 were significantly higher in AERD patients than in ATA patients or HCs (P = .001; respectively). Moreover, levels of serum TGF-β1 and urinary LTE4 had a positive correlation (r = 0.273, P = .037). In the presence of TGF-β1, leukotriene C4 synthase (LTC4S) expression was enhanced in peripheral eosinophils to produce LTE4, which sequentially induced eosinophil degranulation via the p38 pathway. When mice were treated with TGF-β1, significantly induced eosinophilia with increased LTE4 production in the lung tissues were noted. These findings suggest that higher levels of TGF-β1 in AERD patients may contribute to LTE4 production via enhancing LTC4S expression which induces eosinophil degranulation, accelerating airway inflammation.

Topics: Adult; Animals; Aspirin; Asthma, Aspirin-Induced; Eosinophils; Female; Gene Expression Regulation; Glutathione Transferase; Humans; Inflammation; Leukotriene E4; Male; Mice; Middle Aged; p38 Mitogen-Activated Protein Kinases; Receptors, Leukotriene; Respiratory System; Respiratory System Abnormalities; Transforming Growth Factor beta1

Surfactant protein D (SPD) is a member of the collectin family that lines the airway epithelial cells with host defense. However, the role of SPD in the pathogenesis of aspirin-exacerbated respiratory disease (AERD) is still unclear.. The serum SPD level was measured in patients with AERD (n = 336), those with aspirin-tolerant asthma (ATA, n = 442), and healthy controls (HC, n = 104). Polymorphisms of SFTPD in the study subjects were analyzed. The effect of LTE4 on SPD production through eosinophil infiltration was investigated in BALB/c mice. The protective function of SPD against eosinophils inducing inflammation and remodeling was assessed in vitro/vivo. The potential efficacy of nintedanib against airway remodeling through the production of SPD was evaluated.. The decreased level of SPD in AERD was associated with airway inflammation/remodeling under the eosinophilic condition, suggesting that modulation of SPD may provide a potential benefit in AERD.

Topics: Adult; Airway Remodeling; Animals; Asthma, Aspirin-Induced; Eosinophils; Female; Humans; Indoles; Inflammation; Leukotriene E4; Male; Mice; Mice, Inbred BALB C; Middle Aged; Pulmonary Surfactant-Associated Protein D; Respiratory System

Human type-2 CD8

Topics: A549 Cells; Asthma; CD8-Positive T-Lymphocytes; Cell Line, Tumor; Chemokines; Cytokines; Humans; Hypersensitivity; Inflammation; Leukotriene E4; Lipids; Lymphocyte Count; Mast Cells; Prostaglandin D2; Pulmonary Eosinophilia; Th2 Cells

Exposure to low levels of volatile organic compounds (VOCs) in ordinary life is suspected to be related to oxidative stress and decreased lung function. This study evaluated whether exposure to ambient VOCs in indoor air affects airway inflammation.. Thirty-four subjects from the hospital that had moved to a new building were enrolled. Symptoms of sick building syndrome, pulmonary function tests, and fractional exhaled nitric oxide (FeNO) were evaluated, and random urine samples were collected 1 week before and after the move. Urine samples were analyzed for VOC metabolites, oxidative stress biomarkers, and urinary leukotriene E4 (uLTE4) levels.. The level of indoor VOCs in the new building was higher than that in the old building. Symptoms of eye dryness and eye irritation, as well as the level of a xylene metabolite (o-methylhippuric acid) increased after moving into the new building (p = 0.012, p = 0.008, and p < 0.0001, respectively). For the inflammatory markers, FeNO decreased (p = 0.012 and p = 0.04, respectively) and the uLTE4 level increased (p = 0.005) after the move.. Exposure to a higher level of VOCs in everyday life could affect airway inflammation.

Topics: Adult; Aged; Air Pollutants; Biomarkers; Environmental Exposure; Environmental Monitoring; Female; Humans; Inflammation; Leukotriene E4; Male; Middle Aged; Nitric Oxide; Oxidative Stress; Republic of Korea; Respiratory Function Tests; Respiratory System; Sick Building Syndrome; Volatile Organic Compounds

The cysteinyl leukotrienes (cys-LTs), leukotriene C4 (LTC4), a conjugation product of glutathione and eicosatetraenoic acid, and its metabolites, LTD4 and LTE4, are lipid mediators of smooth muscle constriction and inflammation in asthma. LTD4 is the most potent ligand for the type 1 cys-LT receptor (CysLT1R), and LTC4 and LTD4 have similar lesser potency for CysLT2R, whereas LTE4 has little potency for either receptor. Cysltr1/Cysltr2(-/-) mice, lacking the two defined receptors, exhibited a comparable dose-dependent vascular leak to intradermal injection of LTC4 or LTD4 and an augmented response to LTE4 as compared with WT mice. As LTE4 retains a cysteine residue and might provide recognition via a dicarboxylic acid structure, we screened cDNAs within the P2Y nucleotide receptor family containing CysLTRs and dicarboxylic acid receptors with trans-activator reporter gene assays. GPR99, previously described as an oxoglutarate receptor (Oxgr1), showed both a functional and a binding response to LTE4 in these transfectants. We generated Gpr99(-/-) and Gpr99/Cysltr1/Cysltr2(-/-) mice for comparison with WT and Cysltr1/Cysltr2(-/-) mice. Strikingly, GPR99 deficiency in the Cysltr1/Cysltr2(-/-) mice virtually eliminated the vascular leak in response to the cys-LT ligands, indicating GPR99 as a potential CysLT3R active in the Cysltr1/Cysltr2(-/-) mice. Importantly, the Gpr99(-/-) mice showed a dose-dependent loss of LTE4-mediated vascular permeability, but not to LTC4 or LTD4, revealing a preference of GPR99 for LTE4 even when CysLT1R is present. As LTE4 is the predominant cys-LT species in inflamed tissues, GPR99 may provide a new therapeutic target.

Topics: Animals; Capillary Permeability; Inflammation; Leukotriene C4; Leukotriene D4; Leukotriene E4; Ligands; Mice; Mice, Inbred BALB C; Mice, Knockout; Receptors, Leukotriene

Leukotriene E4 (LTE4), the most stable of the cysteinyl leukotrienes (cysLTs), binds poorly to classical type 1 and 2 cysLT receptors although in asthmatic individuals it may potently induce bronchial constriction, airway hyperresponsiveness and inflammatory cell influx to the lung. A recent study has suggested that the purinergic receptor P2Y12 is required for LTE4 mediated pulmonary inflammation in a mouse model of asthma and signals in response to cysLTs. The aim of the study was to characterise the responsiveness of human P2Y12 to cysteinyl leukotrienes. Models of human CysLT1, CysLT2 and P2Y12 overexpressed in HEK293, CHO cells and human platelets were used and responsiveness to different agonists was measured using intracellular calcium, cAMP and β-arrestin recruitment assays. CysLTs induced concentration dependent calcium mobilisation in cells overexpressing CysLT1 and CysLT2 but failed to induce any calcium response in cells expressing P2Y12 or P2Y12+ Gα16. In contrast, selective P2Y12 agonists ADP and 2-MeS-ADP induced specific calcium flux in cells expressing P2Y12+ Gα16. Similarly, specific response to 2-MeS-ADP, but not to cysLTs was also observed in cells expressing P2Y12 when intracellular cAMP and β-arrestin signalling were analysed. Platelets were used as a model of human primary cells expressing P2Y12 to analyse potential signalling and cell activation through P2Y12 receptor or receptor heterodimers but no specific LTE4 responses were observed. These results show that LTE4 as well as other cysLTs do not activate intracellular signalling acting through P2Y12 and suggest that another LTE4 specific receptor has yet to be identified.

Topics: Adenosine Diphosphate; Animals; Arrestins; beta-Arrestins; Blood Platelets; Calcium; CHO Cells; Cricetulus; Cyclic AMP; Cysteine; HEK293 Cells; Humans; Inflammation; Leukotriene E4; Leukotrienes; Mice; Platelet Activation; Purinergic P2Y Receptor Agonists; Receptors, Leukotriene; Receptors, Purinergic P2Y12; Signal Transduction; Thionucleotides

PGD(2) exerts a number of proinflammatory responses through a high-affinity interaction with chemoattractant receptor-homologous molecule expressed on Th2 cells (CRTH2) and has been detected at high concentrations at sites of allergic inflammation. Because cysteinyl leukotrienes (cysLTs) are also produced during the allergic response, we investigated the possibility that cysLTs may modulate the response of human Th2 cells to PGD(2). PGD(2) induced concentration-dependent Th2 cytokine production in the absence of TCR stimulation. Leukotrienes D(4) and E(4) (LTE(4)) also stimulated the cytokine production but were much less active than PGD(2). However, when combined with PGD(2), cysLTs caused a greater than additive enhancement of the response, with LTE(4) being most effective in activating Th2 cells. LTE(4) enhanced calcium mobilization in response to PGD(2) in Th2 cells without affecting endogenous PGD(2) production or CRTH2 receptor expression. The effect of LTE(4) was inhibited by montelukast but not by the P2Y(12) antagonist methylthioadenosine 5'-monophosphate. The enhancing effect was also evident with endogenous cysLTs produced from immunologically activated mast cells because inhibition of cysLT action by montelukast or cysLT synthesis by MK886, an inhibitor of 5-lipoxygenase-activating protein, reduced the response of Th2 cells to the levels produced by PGD(2) alone. These findings reveal that cysLTs, in particular LTE(4), have a significant proinflammatory impact on T cells and demonstrate their effects on Th2 cells are mediated by a montelukast-sensitive receptor.

Topics: Animals; Cells, Cultured; CHO Cells; Cricetinae; Cytokines; Drug Synergism; Humans; Inflammation; Leukotriene E4; Lymphocyte Activation; Mast Cells; Prostaglandin D2; Th2 Cells

Currently, bronchopulmonary dysplasia (BPD) occurs almost exclusively in pre-term infants. In addition to prematurity, other factors like oxygen toxicity and inflammation can contribute to the pathogenesis. This study aimed to compare urinary inflammatory and oxidative stress markers between the no/mild BPD group and moderate/severe BPD group and between BPD cases with significant early lung disease like respiratory distress syndrome (RDS) ('classic' BPD) and with minimal early lung disease ('atypical' BPD). A total of 60 patients who were a gestational age < 30 weeks or a birth weight < 1250 g were included. Urine samples were obtained on the 1(st), 3(rd) and 7(th) day of life and measured the levels of leukotriene E(4) (LTE(4)) and 8-hydroxydeoxyguanosine (8-OHdG). The 8-OHdG values on the 3(rd) day showed significant correlation to duration of mechanical ventilation. The 8-OHdG levels on the 7(th) day were the independent risk factor for developing moderate/severe BPD. In 'classic' BPD, the 8-OHdG values on the 3(rd) day were higher than those of 'atypical' BPD. In 'atypical' BPD, the LTE(4) values on the 7(th) day were higher than the values in 'classic' BPD. These results suggest that oxidative DNA damage could be the crucial mechanism in the pathogenesis of current BPD and the ongoing inflammatory process could be an important mechanism in 'atypical' BPD.

Topics: 8-Hydroxy-2'-Deoxyguanosine; Biomarkers; Bronchopulmonary Dysplasia; Deoxyguanosine; Female; Humans; Infant; Infant, Newborn; Infant, Very Low Birth Weight; Inflammation; Leukotriene E4; Male; Oxidative Stress; Statistics as Topic

Inflammatory environment chronically activates bronchial epithelial cells to stimulate airway cells including epithelial cells themselves by secreting pro-inflammatory and regulatory factors. Proteomic approach is most relevant to screen the epithelial pathways following the inflammatory stimuli. We compared protein expression of the human bronchial epithelial cells exposed to leukotriene E(4) (LTE(4)) and transforming growth factor-beta(1) (TGF-beta(1)) with that of non-stimulated cells. The proteins were separated by 2-DE and the differentially expressed proteins were identified by MALDI-TOF MS and TOF/TOF tandem MS/MS. This approach allowed identification of 31 proteins, of which 26 corresponded to different proteins. beta-tubulin, significantly down-regulated by LTE(4), was confirmed as a ciliated cell marker beta-tubulin IV, whose decrease by LTE(4) was further corroborated by flow cytometry and RT-qPCR. This refers to a contribution of cysteinyl leukotrienes to epithelial remodelling and initiation of epithelial-mesenchymal transition in conducting airways. Of the affected proteins by TGF-beta(1), clinically most relevant ones were up-regulated antioxidant enzyme superoxide dismutase 1, pro-fibrotic enzyme protein disulfide-isomerase and heat shock 70 kDa protein 9B. The changed protein profiles from this study add novel aspects to improve our understanding of the airway pathobiology, provide hints for further directed airway research and may contribute to selecting targets for future therapeutics.

Topics: Bronchi; Electrophoresis, Gel, Two-Dimensional; Epithelial Cells; Flow Cytometry; Gene Expression Regulation; Humans; Inflammation; Leukotriene E4; Protein Disulfide-Isomerases; Proteome; Proteomics; Reverse Transcriptase Polymerase Chain Reaction; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Transforming Growth Factor beta1; Tubulin

Oxidative stress and inflammation have been linked to many chronic diseases including cancer and cardiovascular diseases. Urinary levels of F(2)-isoprostanes (F(2)-IsoPs), 2,3-dinor-5,6-dihydro-15-F(2t)-IsoP (15-F(2t)-IsoP-M), a major metabolite of F(2)-IsoPs, prostaglandin E(2) metabolite (PGE-M), and leukotriene E(4) (LTE(4)) have been proposed as biomarkers for oxidative stress and inflammation. However, little information is available regarding the intra-person variation of these biomarkers, hindering their application in epidemiologic studies.. We evaluated the intra-person variation of these four urinary biomarkers among 48 randomly chosen participants of a validation study of a population-based cohort, the Shanghai Men's Health Study. Four spot urine samples, collected during each season over a 1-year period, were measured for these biomarkers.. The intraclass correlation coefficients for F(2)-IsoPs, 15-F(2t)-IsoP-M, PGE-M, and LTE(4) were 0.69, 0.76, 0.67, and 0.64, respectively. The Spearman correlation coefficients, derived by using bootstrap analysis of single spot measurements and the average of the other three seasonal measurements, were 0.47, 0.60, 0.61, and 0.57 for F(2)-IsoPs, 15-F(2t)-IsoP-M, PGE-M, and LTE(4). Except for high correlations between F(2)-IsoPs and 15-F(2t)-IsoP-M (r = 0.65), the other biomarkers were moderately correlated (r = 0.21-0.44).. Our study results suggest that these four urinary biomarkers have relatively low intra-person variation over a 1-year period.. Spot measurements of F(2)-IsoPs, 15-F(2t)-IsoP-M, PGE-M, and LTE(4) could be useful as biomarkers of oxidative stress and inflammation status for epidemiologic studies.

Topics: Adult; Aged; Biomarkers; Chromatography, Liquid; Cohort Studies; Dinoprostone; F2-Isoprostanes; Gas Chromatography-Mass Spectrometry; Humans; Inflammation; Leukotriene E4; Male; Middle Aged; Oxidative Stress; Tandem Mass Spectrometry

Female hormones play an important role in women's lung health, especially in asthma pathophysiology. Although a growing interest has recently been aroused in asthma related to short-term reproductive states, menopausal asthma has been little studied in the past. The aim of the present study was to explore airway inflammation in menopausal asthmatic women in a noninvasive manner.. Forty consecutive women with menopausal asthma, 35 consecutive women with premenopausal asthma and 30 age-matched healthy controls were enrolled in the study. Urinary LTE-4, induced sputum inflammatory cells, and exhaled LTE-4, IL-6, pH, and NO levels were measured in all the subjects enrolled.. Women with menopausal asthma showed decreased estradiol concentrations, high sputum neutrophils, and exhaled IL-6. Women with premenopausal asthma presented instead an essentially eosinophilic inflammatory pattern. Higher urine and breath condensate LTE-4 concentrations were found in premenopausal and menopausal asthma compared to controls.. Our results substantiate the existence of a new biological phenotype of menopausal asthma that is mainly characterized by neutrophilic airways inflammation and shares several characteristics of the severe asthma phenotype.

Topics: Asthma; Case-Control Studies; Eosinophils; Exhalation; Female; Humans; Hydrogen-Ion Concentration; Inflammation; Interleukin-4; Leukotriene E4; Menopause; Middle Aged; Neutrophils; Nitric Oxide; Phenotype; Premenopause; Sputum

Increased levels of leukotrienes (LTs) in exhaled breath condensate (EBC) are associated with asthma and bronchial hyperresponsiveness (BHR), whereas eicosanoids generated through the 15-lipoxygenase (LO) pathway (15-hydroxyeicosatetraenoic acid [HETE] and eoxins) have been less studied.. We investigated whether metabolites of the 5- and 15-LO pathways in EBC are associated with childhood asthma, asthma severity, and clinical parameters.. The present study included 131 school-aged children (27 children with problematic severe asthma, 80 children with mild-to-moderate asthma, and 24 healthy children) from the Severe Asthma Recognized in Childhood study and 19 children with other nonasthmatic chronic lung diseases. Clinical work-up included spirometry, fractional exhaled nitric oxide measurements, skin prick testing, and methacholine challenge. Eicosanoids were analyzed in EBC by using mass spectrometry and are reported as concentrations (in picograms per milliliter) and eicosanoid/palmitic acid (PA) ratios.. Eoxin C₄/PA, eoxin D₄/PA, eoxin E₄/PA, 15-HETE/PA, and LTC₄/PA ratios were significantly increased in asthmatic versus healthy children. Eoxin D₄/PA and LTE₄/PA ratios were also significantly higher in children with BHR. A nonsignificant trend was observed toward higher eoxin/PA ratios with increasing asthma severity. In contrast to asthma, children with chronic lung disease had the highest 15-HETE/PA, LTC₄/PA, LTE₄/PA, and LTB₄/PA ratios.. The results point to increased activity of the 15-LO inflammatory pathway in childhood asthma. Mass spectrometric analyses of EBC demonstrate that increased eoxin levels not only accompany the increased 5-LO product LTC₄ but are also associated with BHR. These markers might represent a new therapeutic target for asthma treatment.

Topics: Adolescent; Arachidonate 15-Lipoxygenase; Asthma; Breath Tests; Bronchial Hyperreactivity; Child; Exhalation; Female; Humans; Hydroxyeicosatetraenoic Acids; Inflammation; Leukotriene C4; Leukotriene E4; Leukotrienes; Male; Mass Spectrometry; Severity of Illness Index

To assess the clinical significance of three different noninvasive airway inflammatory indices in induced sputum and exhaled breath condensate (EBC) from persistent asthmatic patients.. Moderate and severe asthmatic patients were prescribed inhaled corticosteroids combined with long-acting beta(2) agonists for a month. The symptom scores and percentage of predicted value of forced expiratory volume in one second (FEV(1)) (FEV(1)%pred) were measured while the concentrations of H(2)O(2), NO(3)(-)/NO(2)(-), and cysteinyl-leukotriene E(4) (LTE(4)) in induced sputum and EBC were detected before and after therapy.. A total of twenty-five subjects with moderate and severe asthma were enrolled. By combined therapy for one month the asthma symptoms relieved and FEV(1)%pred improved significantly (P < 0.01). The concentrations of H(2)O(2), NO(3)(-)/NO(2)(-) and LTE(4) in induced sputum and EBC declined significantly (P < 0.01) although the concentrations were still higher than those at normal baseline. More marked reduction of H(2)O(2) and NO(3)(-)/NO(2)(-) compared to LTE(4) was observed. It was revealed that the concentrations of H(2)O(2)and NO(3)(-)/NO(2)(-) but not of LTE(4) in EBC were negatively correlated with FEV(1)%pred (P < 0.01) and positively with symptom scores. Such correlations were also found in H(2)O(2) in induced sputum with FEV(1)%pred and symptom scores as well as NO(3)(-)/NO(2)(-) in induced sputum with FEV(1)%pred. The improvement of FEV(1)%pred after treatment was positively correlated with the reduction of H(2)O(2) and NO(3)(-)/NO(2)(-) both in induced sputum and EBC. Correlation analysis also demonstrated three inflammatory indices were equivalent in induced sputum and EBC (correlation coefficient of H(2)O(2), NO(3)(-)/NO(2)(-) and LTE(4), 0.759, 0.826 and 0.653, respectively. P < 0.01).. (1) Combined therapy with inhaled corticosteroid plus long-acting beta(2) agonist significantly improves the clinical symptoms and lung function of patients with moderate and severe asthma companies with marked suppression of airway inflammation. (2) Both of EBC and induced sputum sampling are valuable noninvasive procedures for detecting asthma airway inflammation, however, EBC technique is superior in safety and reproducibility. (3) H(2)O(2) and NO(3)(-)/NO(2)(-) seem to be more sensitive indices in diagnosis and monitoring asthma compared to LTE(4).

Topics: Adult; Asthma; Biomarkers; Breath Tests; Case-Control Studies; Female; Forced Expiratory Volume; Humans; Hydrogen Peroxide; Inflammation; Leukotriene E4; Male; Middle Aged; Nitric Oxide; Sputum

Cysteinyl leukotrienes (cys-LTs) are potent inflammatory lipid mediators, of which leukotriene (LT) E(4) is the most stable and abundant in vivo. Although only a weak agonist of established G protein-coupled receptors (GPCRs) for cys-LTs, LTE(4) potentiates airway hyper-responsiveness (AHR) by a cyclooxygenase (COX)-dependent mechanism and induces bronchial eosinophilia. We now report that LTE(4) activates human mast cells (MCs) by a pathway involving cooperation between an MK571-sensitive GPCR and peroxisome proliferator-activated receptor (PPAR)gamma, a nuclear receptor for dietary lipids. Although LTD(4) is more potent than LTE(4) for inducing calcium flux by the human MC sarcoma line LAD2, LTE(4) is more potent for inducing proliferation and chemokine generation, and is at least as potent for upregulating COX-2 expression and causing prostaglandin D(2) (PGD(2)) generation. LTE(4) caused phosphorylation of extracellular signal-regulated kinase (ERK), p90RSK, and cyclic AMP-regulated-binding protein (CREB). ERK activation in response to LTE(4), but not to LTD(4), was resistant to inhibitors of phosphoinositol 3-kinase. LTE(4)-mediated COX-2 induction, PGD(2) generation, and ERK phosphorylation were all sensitive to interference by the PPARgamma antagonist GW9662 and to targeted knockdown of PPARgamma. Although LTE(4)-mediated PGD(2) production was also sensitive to MK571, an antagonist for the type 1 receptor for cys-LTs (CysLT(1)R), it was resistant to knockdown of this receptor. This LTE(4)-selective receptor-mediated pathway may explain the unique physiologic responses of human airways to LTE(4) in vivo.

Topics: Cell Proliferation; Chemokines; Dose-Response Relationship, Drug; Enzyme Activation; Extracellular Signal-Regulated MAP Kinases; Humans; Inflammation; Leukemia; Leukotriene D4; Leukotriene E4; Lipids; Mast Cells; Models, Biological; PPAR gamma; Prostaglandin D2

Although eosinophils produce cysteinyl leukotrienes (CysLTs) in large quantities, information on the relationship between CysLTs and eosinophilic pneumonia (EP) is lacking. Inflammatory mediator concentrations in urine were quantified to clarify the relationship between CysLT concentrations and EP severity. Leukotriene (LT)E(4), eosinophil-derived neurotoxin (EDN), 9alpha,11beta-prostaglandin F2 and LTB(4) glucuronide concentrations were quantified in the urine of: EP patients during acute exacerbation and clinical remission; asthmatic patients during acute exacerbation and under stable conditions; and healthy control subjects. The urinary LTE(4) and EDN concentrations of EP patients during acute exacerbation were significantly higher than those of asthmatic patients and healthy subjects, and decreased immediately during clinical remission. The urinary LTE(4) concentration was associated with the urinary EDN concentration of EP patients during acute exacerbation. The urinary LTE(4) concentration significantly correlated with the diffusing capacity of the lung for carbon monoxide in EP patients during acute exacerbation. The increased urinary concentrations of leukotriene and eosinophil-derived neurotoxin were associated with acute exacerbation in eosinophilic pneumonia patients. The increased leukotriene concentration significantly correlated with diffusing capacity of the lung for carbon monoxide, suggesting that the monitoring of leukotriene concentration may aid in the management of eosinophilic pneumonia patients.

Topics: Adolescent; Adult; Aged; Asthma; Case-Control Studies; Female; Glucuronides; Humans; Inflammation; Leukotriene E4; Male; Middle Aged; Neurotoxins; Pulmonary Eosinophilia; Remission Induction

Inhalation of heparin attenuates hyperventilation-induced bronchoconstriction in humans and dogs. The purpose of this study was to determine whether heparin inhibits the late-phase response to hyperventilation, which is characterized by increased peripheral airway resistance (RP), eicosanoid mediator production, neutrophilic/ eosinophilic inflammation, and airway hyperreactivity (AHR) at 5 h after dry air challenge (DAC). Fiberoptic bronchoscopy was used to record RP and airway reactivity (DeltaRP) to aerosol and intravenous histamine before and 5 h after DAC. Bronchoalveolar lavage fluid (BALF) cells and eicosanoid mediators were also measured approximately 5 h after DAC. DAC of vehicle-treated bronchi resulted in late-phase airway obstruction (approximately 120% increase over baseline RP), inflammation, increased BALF concentrations of leukotriene (LT) C(4), LTD(4), and LTE(4) and prostaglandin (PG)D(2), and AHR. Pretreatment with aerosolized heparin attenuated late-phase airway obstruction by approximately 50%, inhibited eosinophil infiltration, reduced BALF concentrations of LTC(4), LTD(4), and LTE(4) and PGD(2), and abolished AHR. We conclude that heparin inhibits hyperventilation-induced late-phase changes in peripheral airway function, and does so in part via the inhibition of eosinophil migration and eicosanoid mediator production and release.

Topics: Administration, Inhalation; Airway Resistance; Animals; Bronchial Hyperreactivity; Bronchial Provocation Tests; Bronchoalveolar Lavage Fluid; Bronchoscopy; Disease Models, Animal; Dogs; Drug Evaluation, Preclinical; Eicosanoids; Eosinophils; Heparin; Humans; Hyperventilation; Inflammation; Leukotriene C4; Leukotriene D4; Leukotriene E4; Male; Neutrophils; Prostaglandin D2; Time Factors

Topics: Acute Disease; Animals; Blood Platelets; Disease Models, Animal; In Vitro Techniques; Inflammation; Kinetics; Leukocyte Count; Leukocytes, Mononuclear; Leukotriene B4; Leukotriene E4; Male; N-Formylmethionine Leucyl-Phenylalanine; Neutrophils; Platelet Count; Rabbits

Bleomycin and asparaginase are widely used antineoplastic agents which may induce allergic or inflammatory side-effects. Mast cells are implicated as effector cells in allergic and inflammatory responses. The aim of this study was to establish whether bleomycin or asparaginase modulate leukotriene production in vitro and in vivo. Leukotriene C4 (LTC4) production by murine bone marrow-derived mast cells (BMMC) was determined by radioimmunoassay (RIA). Leukotriene production in patients was assessed by determining leukotriene E4 and N-acetyl-leukotriene E4 in urine by means of combined HPLC and RIA. Bleomycin induced an up to 2.1-fold increase in LTC4 production both in unstimulated and in calcium ionophore-stimulated mast cells. In 3 of 7 patients treated with bleomycin, a greater than 2-fold increase in endogenous leukotriene production was observed. This effect was associated with febrile responses and was most pronounced in a patient who developed an Adult Respiratory Distress Syndrome (ARDS). Asparaginase increased leukotriene production up to 10-fold in stimulated but not in unstimulated BMMC. In a patient who developed an anaphylactic reaction after treatment with asparaginase, a pronounced increase in urinary leukotriene concentration was observed. In contrast to bleomycin or asparaginase, a number of other cytostatic agents did not significantly change leukotriene production by BMMC. Our data indicate that some of the inflammatory and allergic side-effects of bleomycin and asparaginase could be mediated by leukotrienes, a possible source of which may be mast cells.

Topics: Adult; Anaphylaxis; Animals; Antineoplastic Agents; Asparaginase; Bleomycin; Calcimycin; Drug Hypersensitivity; Humans; In Vitro Techniques; Inflammation; Ionophores; Leukotriene C4; Leukotriene E4; Leukotrienes; Lymphoma, Non-Hodgkin; Mast Cells; Mice; Mice, Inbred BALB C; Respiratory Distress Syndrome

Myrtol standardized (Gelomyrtol/Gelomyrtol forte) inhibits the activity of 5-lipoxygenase of human basophil and eosinophil leukocytes and the formation of leukotriene C4 as well as 1.8-cineole (eucalyptol). An increase of prostaglandins (PGE2) in mucous membranes of teat cisterns after topical administration of TPA (tetradecanoylphorbol-13-acetate) was inhibited. In vitro and in vivo studies revealed spasmolytic and broncholytic effects of Myrtol stand. After topical administration into the teat cisterns of the isolated bovine udder Myrtol stand. increased the surface temperature, comparable to effects of menthol.

Topics: Animals; Arachidonate 5-Lipoxygenase; Arachidonic Acid; Basophils; Cattle; Cyclohexanols; Dinoprostone; Drug Combinations; Drug Interactions; Eosinophils; Eucalyptol; Female; Guinea Pigs; Humans; Hyperemia; In Vitro Techniques; Inflammation; Leukemia, Basophilic, Acute; Leukotriene C4; Leukotriene D4; Leukotriene E4; Male; Mammary Glands, Animal; Menthol; Mice; Monoterpenes; Mucous Membrane; Muscle Contraction; Muscle, Smooth; Rats; Terpenes; Tetradecanoylphorbol Acetate; Trachea; Tumor Cells, Cultured

Inflammation may contribute to the pathogenesis of high-altitude pulmonary edema (HAPE). This study was designed to determine whether a marker of inflammation, urinary leukotriene E4 (LTE4), is elevated in patients with HAPE.. We conducted a case-control study to collect clinical data and urine samples from HAPE patients and healthy control subjects at moderate altitude (> or = 2727 m), and follow-up urine samples from HAPE patients following their return to low altitude (< or = 1,600 m).. Five medical clinics in Summit County, Colorado.. Questionnaire data were evaluated in 71 HAPE patients and 36 control subjects. Urinary LTE4 levels were determined from a random subset of 38 HAPE patients and 10 control subjects presenting at moderate altitude, and on 5 HAPE patients who had returned to low altitude.. Using an enzyme immunoassay technique, urinary LTE4 levels were found to be significantly higher in HAPE patients (123 [16 to 468] pg/mg creatinine, geometric mean [range]) than in control subjects (69 [38 to 135]), p = 0.02. Following return to low altitude, urinary LTE4 levels fell significantly from 122 (41.8 to 309) to 53.6 (27.6 to 104) pg/mg creatinine (p = 0.05). Urinary LTE4 levels were not related to age, sex, time at altitude, physical condition or habitual exercise, recent use of alcohol or nonsteroidal anti-inflammatory drugs (NSAIDs), or oxygen saturation. Clinical factors associated with HAPE included male sex, regular exercise, and recent use of NSAIDs.. We conclude that urinary LTE4 levels are elevated in patients with HAPE, supporting the view that HAPE involves inflammatory mechanisms.

Topics: Adult; Altitude; Case-Control Studies; Creatinine; Female; Humans; Immunoenzyme Techniques; Inflammation; Leukotriene E4; Male; Middle Aged; Pulmonary Edema; Risk Factors

We used the 5-lipoxygenase-activating protein (FLAP) antagonist MK-0591 to investigate the importance of leukotrienes (LT) in causing ozone-induced bronchoconstriction, airway inflammation, and airway hyperresponsiveness in dogs. Six random source dogs were studied. On one day, dogs were treated with MK-0591 (2 mg/kg iv) followed by a continuous intravenous infusion of 8 micrograms.kg-1.min-1. On the other day, the diluent was infused. Acetylcholine airway responsiveness was measured before and 1 h after ozone inhalation (3 ppm for 30 min). On each day, whole blood and bronchoalveolar lavage (BAL) cells were challenged with calcium ionophore to stimulate LTB4 production. Urinary LTE4 levels were measured before and after ozone. MK-0591 inhibited LTB4 production in whole blood by 96% (P = 0.001) and that from BAL cells by 91% (P = 0.001). By contrast, MK-0591 had no effect on ozone-induced bronchoconstriction, airway hyperresponsiveness, or influx of neutrophils into BAL. The mean log difference of the pre- to post-acetylcholine provocative concentration was 0.64 +/- 0.40 during MK-0591 treatment and 0.68 +/- 0.40 during diluent treatment (P = 0.71). These results indicate that peptidoleukotrienes are produced during ozone inhalation and that MK-0591 inhibits LT production in dogs. However, LTs do not play a role in ozone-induced bronchoconstriction, airway inflammation, or airway hyperresponsiveness in dogs.

Topics: 5-Lipoxygenase-Activating Proteins; Acetylcholine; Airway Resistance; Animals; Bronchial Hyperreactivity; Bronchoalveolar Lavage Fluid; Carrier Proteins; Dogs; Indoles; Inflammation; Leukotriene Antagonists; Leukotriene B4; Leukotriene E4; Leukotrienes; Lipoxygenase Inhibitors; Membrane Proteins; Ozone; Quinolines; Respiratory System

Sickle cell (HbSS) disease is associated with rheological and inflammatory stresses within the microcirculation. In order to determine the role of leukotrienes in the inflammatory processes in HbSS patients, we analysed plasma and urine levels of leukotrienes (LT); LTB4, LTC4, LTD4, and LTE4 as indicators of their in vivo metabolism. Plasma and urine level samples of 15 HbSS patients in steady-state and age-matched healthy, homozygous (HbAA) controls were extracted for leukotrienes and quantitated by HPLC. Control plasma level of leukotrienes (mean +/- SEM, ng ml-1) were: LTB4, 8.95 +/- 0.26; LTC4, 7.24 +/- 0.21; LTD4, 11.42 +/- 0.40; and LTE4, 14.51 +/- 0.50. Corresponding values for HbSS patients were: LTB4, 6.15 +/- 0.42; LTC4, 13.61 +/- 1.45; LTD4, 6.44 +/- 0.51 and LTE4, 4.97 +/- 0.37. The differences were significant at P < 0.05. Urine levels (mean +/- SEM, ng mmol-1 creatinine), for controls were: LTB4, 10.60 +/- 0.35; LTC4, 360.0 +/- 9.82. Values for HbSS urine were: LTB4, 27.50 +/- 3.33; LTC4, 356.0 +/- 17.87; LTD4, 69.90 +/- 14.51. LTD4 was not detected in control urine. These results suggest that sickle cell patients may exhibit impaired ability to catabolize LTC4 in plasma during steady state conditions. This altered metabolism may contribute to the persistent stress of the microcirculation, and is probably related to the abnormal microvascular rheology of sickle blood cells.

Topics: Adult; Anemia, Sickle Cell; Cell Adhesion; Endothelium, Vascular; Female; Humans; Inflammation; Leukocytes; Leukotriene B4; Leukotriene C4; Leukotriene D4; Leukotriene E4; Leukotrienes; Male; Middle Aged

Increased synthesis of peptidoleukotrienes may occur in a variety of inflammatory diseases. To test this theory, hospitalized patients with a variety of diseases were studied and urine LTE4 quantitated as an index of total body peptidoleukotriene synthesis. 10 patients with ARDS, 7 of which had additional organ involvement, and 5 patients suffering from severe burn injuries were studied. Patients with uncomplicated ARDS excreted approximately 6-fold higher amounts of LTE4 in urine compared to healthy subjects. When ARDS was complicated by multiple organ failure (MOF), urine LTE4 levels were 2- to 150-fold higher than in healthy volunteers. Patients with severe burn injuries had peak urine LTE4 levels which were approximately 20-fold higher than in healthy volunteers. As additional controls, patients with cardiac arrhythmias (absence of inflammatory disease) and patients with uncomplicated pneumonia (localized inflammation) showed normal or mildly elevated urinary LTE4 levels. The urinary LTE4 levels in ARDS patients did not correlate with serum creatinine, bilirubin, or LDH levels, or with the WBC, nor did renal or liver failure by itself predict extremely elevated urinary LTE4 levels. In conclusion, patients with ARDS or ARDS/MOF and patients with severe injuries and sepsis syndrome excrete higher levels of urinary LTE4 than patients healthy volunteers or patients with limited inflammatory disease. In certain situations, urinary LTE4 levels may be useful as a marker of the degree of inflammation.

Topics: Burns; Chromatography, High Pressure Liquid; Humans; Inflammation; Intensive Care Units; Leukotriene E4; Multiple Organ Failure; Respiratory Distress Syndrome; SRS-A; Tritium

Using an allergic inflammation model of air pouch type in rats, levels of peptide-leukotriene (LT) C4, D4 and E4 in the pouch fluid were measured chromatographically, and peptide-LT metabolizing activities in the pouch fluid in the anaphylactic phase were examined. 10 min after injection of an antigen (azobenzene arsonate-conjugated acetyl bovine serum albumin) solution into a preformed air pouch on the back of the immunized rats, LTC4 level in the pouch fluid was the highest, followed by LTD4 and LTE4. At 30 min, the order of the level was reversed to LTE4 greater than LTD4 greater than LTC4, and total amount of peptide-LTs (LTC4 + LTD4 + LTE4) was the highest. Supernatant fraction of the pouch fluid collected 30 min after the antigenic challenge, converted [3H]LTC4 into [3H]LTD4, and [3H]LTD4 into [3H]LTE4 in time- and concentration-dependent manner. [3H]LTE4 was not metabolized under these conditions. Heat denaturation of the pouch fluid diminished the conversion of [3H]LTC4 into [3H]LTD4, and [3H]LTD4 into [3H]LTE4. In the granule fraction of purified mast cells, no metabolic activity of [3H]LTs was found. In intact mast cells as well as degranulating mast cells, a small but significant amount of [3H]LTC4 was metabolized into [3H]LTD4 and [3H]LTE4. In contrast, rat serum showed potent metabolizing activities of peptide-LTs. Since plasma exudation into the pouch is very prominent in the anaphylactic phase in this model, peptide-LT metabolizing activities in the pouch fluid are suggested to be attributable to plasma leaked into the pouch during the anaphylactic phase.

Topics: Anaphylaxis; Animals; Hypersensitivity; Inflammation; Leukotriene E4; Male; Mast Cells; Rats; Rats, Inbred Strains; SRS-A

Intraperitoneal injection of zymosan in mice induced rapid extravasation and accumulation of plasma protein in the peritoneal cavity. Neutrophils began to appear in the peritoneal cavity after a lag period of approximately 3 hours. The injected mice exhibited a pain response (writhing) during the first 30 minutes after injection, but writhing ceased before protein or cell accumulation had reached maximum levels. The injection of zymosan induced synthesis of PGE2 (measured by RIA) which reached maximum levels at 30 minutes, then declined slowly. Peptido-leukotriene levels (detected by bioassay, RIA and HPLC) increased rapidly after injection, reached a peak within an hour of injection and declined to undetectable levels within 4 hours. The early peptido-LT was predominantly LTC4, while later, LTE4 was the major component. LTD4 levels remained low throughout and no LTB4 was detected at any time. Indomethacin treatment elevated levels of peptido-LTs, reduced PGE2 levels and inhibited writhing. Phenidone reduced peptido-LT levels. In vitro studies demonstrated that zymosan stimulates LTC4 synthesis by peritoneal cells whereas LTE4, LTD4, LTB4 or monoHETES were not detectable (using HPLC methods). The source of enzymes responsible for the in vivo metabolism of LTC4 to LTD4 and LTE4 could not be identified.

Topics: Animals; Arachidonate Lipoxygenases; Cell Aggregation; Dinoprostone; In Vitro Techniques; Indomethacin; Inflammation; Injections, Intraperitoneal; Leukotriene B4; Leukotriene E4; Lipoxygenase; Male; Mice; Mice, Inbred Strains; Pain; Prostaglandins E; Proteins; SRS-A; Zymosan

Topics: Arachidonic Acids; Asthma; Humans; Hypersensitivity; Inflammation; Leukotriene A4; Leukotriene B4; Leukotriene E4; SRS-A