interleukin-8 and Sinusitis

Topics: Chronic Disease; Humans; Interleukin-8; Nasal Mucosa; Nasal Polyps; Rhinitis; Sinusitis

Chronic sinusitis is a common inflammatory upper respiratory tract disease. One of the prominent features of this disease is persistent purulent effusion containing numerous emigrated neutrophils in the paranasal sinuses. Recent advances in sinusitis research have revealed two positive feedback mechanisms that explain the chronic neutrophil accumulation in the sinus. First, interleukin (IL)-1beta secreted by monocytes, macrophages and fibroblasts upregulates the expression of E-selectin and intercellular adhesion molecule-1 (ICAM-1) in vascular endothelial cells, and thereby induces the extravascular transmigration of neutrophils. The emigrated neutrophils then secrete IL-1beta, which amplifies the expression of E-selectin and ICAM-1, resulting in further neutrophil infiltration. Second, chemoattractants including IL-8 in the sinus effusion initiate neutrophil exudation. Emigrated neutrophils then secrete IL-8, which elicits further neutrophil accumulation in the sinus effusion. Long-term low-dose macrolide therapy was first introduced for the treatment of diffuse panbronchiolitis in the 1980's. In the 1990's it was also shown to be an effective treatment for chronic sinusitis. The inhibitory effect of macrolides on neutrophil infiltration in inflammatory sites has been well documented in these diseases. Several lines of evidence indicate that macrolides do not function simply as a bactericide. In vitro studies have demonstrated various effects of macrolides on immunocompetent cells, inflammatory cells and airway epithelial cells. It has been shown that macrolides inhibit the production of IL-8 and IL-1beta and the expression of ICAM-1, suggesting that macrolides block the aforementioned dual positive feedback system of neutrophil recruitment and thereby exert their clinical efficacy in the treatment of chronic sinusitis. The inhibitory effects of macrolides on multiple steps in the process of neutrophil recruitment are presumably mediated by the inhibition of transcription factors such as nuclear factor-kB and activator protein-1. Further investigation of the mode of action of macrolides at the molecular level would lead to the development of safer and more effective drugs for the treatment of chronic sinusitis. In addition, the possible risk of this therapy such as the occurrence of resistant strains have to be carefully surveyed hereafter.

Topics: Animals; Anti-Bacterial Agents; Chronic Disease; Humans; Intercellular Adhesion Molecule-1; Interleukin-1; Interleukin-8; Macrolides; Neutrophils; Sinusitis

Apart from ventilatory and bacteriologic aspects, understanding the pathomechanisms of inflammation in chronic sinusitis and nasal polyposis seems crucial for further success in disease treatment. New insights into inflammatory processes became recently possible by investigating the pattern of cytokines and chemokines as well as adhesion molecules in different acute and chronic sinus diseases. The proinflammatory cytokines interleukin (IL)-1 beta, IL-6 and especially the neutrophil-chemoattractant IL-8 play a dominant role in acute sinusitis, as was shown before for viral and allergic rhinitis. In contrast, IL-3 protein dominates the cytokine profile in chronic sinusitis, giving support to a variety of inflammatory cells. The most striking finding was the increased synthesis of IL-5 protein in bilateral nasal polyposis, whereas IL-5 was not found in controls or antrochoanal polyps. As this cytokine is known to enhance eosinophil activation and survival, our data point to IL-5 as a key protein in the pathomechanism of tissue eosinophilia in nasal polyposis. The investigation of cytokine patterns may furthermore help to differentiate between sinusitis subgroups, e.g. in the classification of sinus diseases.

Topics: Acute Disease; Cell Adhesion Molecules; Chemokines; Chemotaxis, Leukocyte; Chronic Disease; Cytokines; Eosinophilia; Eosinophils; Humans; Interleukin-1; Interleukin-3; Interleukin-5; Interleukin-6; Interleukin-8; Maxillary Sinus; Nasal Polyps; Neutrophils; Paranasal Sinus Neoplasms; Polyps; Rhinitis; Sinusitis

Low-dose clarithromycin has been recommended for the treatment of chronic rhinosinusitis without nasal polyps. However, it is uncertain whether a high dose of clarithromycin is more effective than a low dose.. Forty-three chronic rhinosinusitis patients were randomised to low-dose or high-dose clarithromycin groups, and clinical efficacy was evaluated. Pre- and post-treatment measures included: nasal symptom assessment, endoscopic inspection (Lund-Kennedy system), a quality of life questionnaire (the Sino-Nasal Outcome Test 20) and examination of cytokine levels (interleukin-5 and -8) in nasal secretions.. The high dose of clarithromycin was significantly better in terms of clinical efficacy than the low dose for the treatment of chronic rhinosinusitis (p < 0.025). Significant differences in nasal cytokine levels (interleukin-5 and -8) were also observed between the low-dose and high-dose groups after short-term clarithromycin treatment (p < 0.025).. Short-term, high-dose clarithromycin appears to be more effective for the treatment of chronic rhinosinusitis than low-dose clarithromycin.

Topics: Adult; Anti-Bacterial Agents; Biomarkers; Chronic Disease; Clarithromycin; Dose-Response Relationship, Drug; Female; Humans; Interleukin-5; Interleukin-8; Male; Middle Aged; Quality of Life; Rhinitis; Sinusitis; Surveys and Questionnaires; Treatment Outcome

To explore the correlation between the expression of GATA-3 and the level of local cytokines (IL-5, IL-6 and IL-8).. The levels of IL-5, IL-6 and IL-8 in ethmoid sinus mucosa were titrated in 45 patients with chronic rhinosinusitis and 11 normal subjects by ELISA. Patients were divided into AR group (with allergic rhinitis) and NAR group (without allergic rhinitis) . Semi-quantitative RT-PCR and immunohistochemical staining were used to examine the GATA-3 expression in nasal mucosa. The correlation between the expression of GATA-3 and the levels of cytokines was evaluated.. IL-5, IL-6 and IL-8 levels in both AR and NAR groups were significantly elevated compared with normal group (all P < 0.01 for AR group; P < 0.05, 0.05, 0.01 for NAR group, respectively), and they were much higher in AR group in comparison with NAR group (P < 0.01, 0.05, 0.01, respectively). Semi-quantitative RT-PCR showed that AR and NAR groups had markedly greater level of GATA-3 mRNA than that in control group (P < 0.01, respectively), and the level of GATA-3 mRNA in AR group was further higher than that in NAR group (P < 0.01). Immunohistochemical staining illustrated that GATA-3 was primarily presented in cytoplasma and the GATA-3 positive cells were mainly infiltrating inflammatory cells in submucosa. The mean GATA-3 positive-staining rate was (27. 90 +/- 16.75)% and (10.22 +/- 0.05)% in AR and NAR group, which were markedly higher than (1.30 +/- 1.78)% in control group (P < 0.01, respectively). Pearson correlation analysis demonstrated that GATA-3 positive-staining rate was closely correlated with IL-5 level, but not IL-6 and IL-8. The correlation coefficient was 0. 712 for GATA-3 and IL-5 (P < 0.01), 0.200 for GATA-3 and IL-6 (P > 0.05), 0.089 for GATA-3 and IL-8 (P > 0.05).. Activation of GATA-3 might be one of the mechanisms for induction of IL-5 expression in chronic rhinosinusitis . Concomitance of allergic rhinitis with chronic rhinosinusitis further increased expression of GATA-3, and subsequently enhanced IL-5 expression. Chronic sinusitis may be related to allergy, and GATA-3 may play a key role in the pathogenesis of chronic sinusitis.

Topics: Adolescent; Adult; Aged; Chronic Disease; Female; GATA3 Transcription Factor; Humans; Interleukin-5; Interleukin-6; Interleukin-8; Male; Middle Aged; Nasal Mucosa; Sinusitis; Young Adult

Recently, the YAMIK sinus catheter (YAMIK) has been reported to be a useful therapeutic device in the treatment of sinusitis. The present study was conducted to compare its delivery of either a normal saline (NS) or a betamethasone solution (0.4 mg/ml) into the paranasal sinuses of 25 patients (39 sides) with chronic sinusitis. The following parameters were evaluated: (1) subjective nasal clinical symptoms (nasal discharge, nasal obstruction, postnasal drip and headache), (2) X-ray photographs (ethmoid and maxillary sinuses) and (3) cytokine levels (IL-1beta, IL-8 and TNF-alpha) by enzyme-linked immunosorbent assay. The total nasal symptom scores significantly decreased after the first therapy, and the total X-ray photograph scores significantly decreased after therapy with either NS or the betamethasone solution. In both NS and betamethasone patients, the levels of IL-1beta and IL-8 had significantly decreased by the 3rd and 2nd weeks after therapy, respectively. In contrast, the TNF-alpha level decreased after the first therapy with betamethasone solution and remained unchanged after therapy with NS. These findings suggest that evacuation of the pathological effusions in sinuses may exert a beneficial effect by reducing the levels of IL-1beta and IL-8, and we speculate that removal of pathological effusions from the sinuses may provide treatment through different mechanisms than those that occur in treatment with betamethasone.

Topics: Adult; Aged; Anti-Inflammatory Agents; Betamethasone; Catheterization; Chronic Disease; Enzyme-Linked Immunosorbent Assay; Female; Humans; Interleukin-1; Interleukin-8; Male; Middle Aged; Sinusitis; Tumor Necrosis Factor-alpha

Chronic refractory sinusitis is a common feature in patients with primary immunodeficiencies. The efficacy of standard therapeutic strategies is questionable. In an open trial we evaluated the efficacy of azithromycin, N-acetylcysteine and topical intranasal beclomethasone (100 microg twice daily for 6 weeks) in 16 patients with primary immunodeficiencies (median age 13.5 years, range 5-32 years). All patients suffered from chronic sinusitis despite regular immunoglobulin replacement therapy every 3 weeks. Magnetic resonance imaging (MRI) scans were performed before and after 6 weeks of treatment to evaluate morphological changes in the paranasal sinuses. Nasal swabs and washings were taken for microbial analysis and measurement of inflammatory mediators (IL-8, tumour necrosis factor-alpha (TNF-alpha), eosinophilic cationic protein (ECP)) before and post therapy. Inflammatory mediators in nasal secretions were significantly elevated in patients: IL-8 median 2436 pg/ml (range 441-5435 pg/ml), TNF-alpha 37.3 pg/ml (3.75-524 pg/ml) and ECP 33 ng/ml (1.5-250 ng/ml) versus age-matched healthy controls: IL-8 median 212 pg/ml (99-825 pg/ml), TNF-alpha 3.77 pg/ml (2.8-10.2 pg/ml) and ECP 1.5 ng/ml (1.5-14.8 ng/ml) (P < 0.0001). Inflammation of the maxillary sinuses was confirmed by MRI scans in all patients, additionally infection of the ethmoidal and frontal sinuses was recorded in five patients. Bacterial growth appeared in 11 out of 16 cultures. In spite of therapy, no improvement in sinal inflammation visualized by MRI was achieved. Moreover, no significant decrease in pathogens and levels of inflammatory mediators could be detected (IL-8 1141 pg/ml, 426-4556 pg/ml; TNF-alpha 13.9 pg/ml, 4.1-291.6 pg/ml; ECP 32.3 ng/ml, 3.7-58.4 ng/ml). Our results demonstrate that conventional management of sinusitis is of little benefit in patients with chronic refractory sinusitis with an underlying immunodeficiency. More studies are needed to test antibiotic regimens, probably combined with surgical drainage and anti-inflammatory agents.

Topics: Acetylcysteine; Administration, Intranasal; Adolescent; Adult; Agammaglobulinemia; Anti-Bacterial Agents; Anti-Inflammatory Agents; Antiviral Agents; Ataxia Telangiectasia; Azithromycin; Beclomethasone; Blood Proteins; Child; Child, Preschool; Chronic Disease; Common Variable Immunodeficiency; Eosinophil Granule Proteins; Ethmoid Sinusitis; Female; Frontal Sinusitis; Glucocorticoids; Humans; Immunization, Passive; Interleukin-8; Lymphoproliferative Disorders; Magnetic Resonance Imaging; Male; Nasal Lavage Fluid; Paranasal Sinuses; Radiography; Ribonucleases; Sinusitis; Tumor Necrosis Factor-alpha

To examine whether and how interleukin (IL)-1α is involved in chronic rhinosinusitis with nasal polyps (CRSwNP).. Nasal polyp (NP) and control tissues were collected from CRSwNP patients and control subjects. The expression of IL-1α and other proinflammatory cytokines (IL-1β, IL-8 and IL-13, etc.), as well as neutrophil and eosinophil accumulation, were examined in sinonasal tissues using immunohistochemical (IHC), immunofluorescent (IF) staining, qPCR, and Luminex, respectively. Moreover, the regulation of IL-1α expression and its effects on other proinflammatory cytokines were evaluated in cultured nasal epithelial cells (NECs).. The mRNA and protein levels of IL-1α were significantly higher in NP tissues compared to that in control tissues. IL-1α in polyp tissues was mainly located in epithelial cells and neutrophils. Polyps IL-1α level was significantly associated with IL-8, IL-1β, IL-6, IL-4 and IL-13 production, as well as tissue neutrophil infiltration. Moreover, poly (I:C), lipopolysaccharides, Flagellin, R848 and cytokines (IL-4, IL-5, and IL-13) significantly increased the expression of IL-1α in cultured NECs in vitro, and recombinant IL-1α significantly promoted production of IL-8 and CXCL1 in cultured NECs.. These findings provided the evidence that IL-1α were significantly increased in NP tissues, which may contribute to tissue neutrophilia in CRSwNP patients in China.

Topics: Chronic Disease; Humans; Interleukin-13; Interleukin-4; Interleukin-8; Nasal Polyps; Rhinitis; Sinusitis

Eosinophilic chronic rhinosinusitis (ECRS) is a subtype of chronic rhinosinusitis (CRS) and is a refractory or intractable disease. However, a reliable clinical marker or an effective treatment strategy has not yet been established. ECRS is accompanied by excessive eosinophil infiltration and Th2 inflammatory response, which is closely related to tissue remodeling in the upper airways.. We sought to investigate the effect of eosinophils on tissue remodeling in ECRS. The purpose of this study was to identify the effects of eosinophils on the expression of pro-inflammatory mediators and extracellular matrix (ECM) in nasal fibroblasts and the key mediators that stimulate them.. Butyric acid was used to differentiate EOL-1 cells into eosinophils. We co-cultured differentiated EOL-1 cells and fibroblasts to measure the expression of pro-inflammatory mediators and ECM in fibroblasts. Among the cytokines secreted from the differentiated EOL-1 cells, factors that induced tissue remodeling of fibroblasts were identified.. Treatment with butyric acid (BA) differentiated EOL-1 cells into eosinophils. Differentiated EOL-1 cells induced fibroblasts to produce pro-inflammatory mediators, IL-6 and IL-8, and tissue remodeling factor, VEGF. It also induced myofibroblast differentiation and overexpression of ECM components. Differentiated EOL-1 cells overexpressed osteopontin (OPN), and recombinant OPN increased the expression of IL-6, IL-8, VEGF, and ECM components in nasal fibroblast. OPN was overexpressed in the nasal tissue of patients with ECRS and was associated with the severity of CRS.. Eosinophil-derived OPN stimulated nasal fibroblasts and contributed to inflammation and tissue remodeling in ECRS. Moreover, the expression level of OPN was proportional to the severity of ECRS. Therefore, OPN regulation is a potential treatment for ECRS.

Topics: Butyric Acid; Chronic Disease; Eosinophils; Extracellular Matrix; Fibroblasts; Humans; Inflammation Mediators; Interleukin-6; Interleukin-8; Nasal Polyps; Osteopontin; Sinusitis; Vascular Endothelial Growth Factor A

Although metabolomics provides novel insights into disease mechanisms and biomarkers, the metabolic alterations in local tissues affected by chronic rhinosinusitis (CRS) are unknown.. This study aimed to determine the metabolomic profiles of sinonasal tissues associated with different types of CRS and their treatment outcomes.. Untargeted metabolomic profiling was performed on sinonasal tissues obtained from patients with eosinophilic CRS with nasal polyps (CRSwNP), noneosinophilic CRSwNP or CRS without nasal polyps (CRSsNP), and controls. The messenger RNA (mRNA) levels of inflammatory cytokines in nasal tissues were detected by quantitative real-time reverse transcriptase PCR. Nasal polyp tissues were cultured ex vivo and treated with glutathione.. Distinct metabolomic profiles were observed for the CRS subtypes. Eosinophilic CRSwNP had profoundly enhanced unsaturated fatty acid oxidization, which correlated with mucosal eosinophil numbers and IL-5 mRNA levels. Noneosinophilic CRSwNP was characterized by uric acid accumulation. Increased uric acid levels were positively correlated with mucosal neutrophil numbers and IFN-γ, IL-17A, IL-1β, and IL-8 mRNA levels. Disrupted purine metabolism was specifically detected in CRSsNP. Reduced levels of amino acid metabolites were found in eosinophilic CRSwNP and CRSsNP, and were inversely associated with mucosal total inflammatory cell numbers and inflammatory cytokines. Compared to non-difficult-to-treat CRS, difficult-to-treat CRS had higher glutathione disulfide levels, which were positively correlated with IL-8 mRNA levels. Glutathione treatment reduced IL-8 mRNA expression in cultured nasal polyp tissues.. Specific metabolic signatures are associated with different types of CRS, inflammatory patterns, and disease outcomes, which may provide novel insights into pathophysiologic mechanisms, subtype-specific biomarkers, and treatment targets of CRS.

Topics: Biomarkers; Chronic Disease; Cytokines; Glutathione; Humans; Interleukin-8; Nasal Polyps; Rhinitis; RNA, Messenger; Sinusitis; Uric Acid

This study was to explore the mechanism of action of nanomaterial-loaded clarithromycin (CLA) after sinusitis surgery. Under the guidance of dynamic enhanced scanning (DES). 120 patients with sinusitis admitted to the First Affiliated Hospital of China Medical University from July 2019 to March 2020 were selected and divided into a control group and an observation group according to the random number table method, with 60 cases in each group. Then, the CLA-containing nano-polylactic acid material was prepared, observed with the scanning electron microscope (SEM), and its drug release ability was tested. All patients underwent endoscopic sinus surgery under general anesthesia. After the surgery was completed, patients in the control group were given only CLA capsules, and patients in the observation group were given freshly prepared nanomaterial-loaded CLA, and both groups of patients were continuously observed for two weeks. After that, the patients were examined using the dynamic enhancement computed tomography (CT). The clinical efficacy, serum interleukin-4 (IL-4), interleukin-8 (IL-8), and tumor necrosis factor α (TNF-α) levels of the two groups of patients were observed. The secretions of the patients' sinuses were performed with microbial bacterial culture, and the results were observed and recorded. Results showed that the characterization and analysis of the nano drug-carrying preparation suggested that the polylactic acid nanomembrane showed linear fiber morphology, relatively dense distribution, not greatly different fiber diameter, and small porosity. Characterization under a field of view (FOV) of 500 um showed that the fiber surface was smooth and rich in content. The release of CLA showed a gradual and steady upward trend. On the 25th day, nearly 50% of the dose had been released, and it had reached more than 90% of the total release on the 55th day. According to the statistics on the clinical efficacy of patients, it was found that the number of cured and effective patients in the observation group was higher than that of the control group, while the number of ineffective cases was much lower than that of the control group. The dynamic enhanced CT examination results of the patients in the control group after treatment showed that the soft tissue mass on the posterior right side of the nasopharynx was reduced, but the pharyngeal suture still existed; while those in the observation group showed that the plain scan density was uniform, and th

Topics: Clarithromycin; Humans; Interleukin-4; Interleukin-8; Nanoparticle Drug Delivery System; Nanostructures; Sinusitis; Staphylococcus aureus; Tumor Necrosis Factor-alpha

Chronic rhinosinusitis with nasal polyps (CRSwNP) is a chronic inflammation of the mucosa of the nose and paranasal sinuses with the presence of polyps, affecting between 2.7% and 4.4% of the population. Cytokine analysis has become important in research on inflammatory mechanisms in CRSwNP. Therefore, our aim is to investigate the complex appearance, relative distribution, and interlinks of IL-1, IL-4, IL-6, IL-7, IL-8, IL-10, IL-12, and Ki-67 in CRSwNP.. Samples of nasal polyps were obtained from 19 patients with previously diagnosed CRSwNP and the recurrence of polyps after previous surgeries. The control group consisted of samples from 17 otherwise healthy individuals with isolated nasal septum deviations. Tissues were stained for previously mentioned cytokines and Ki-67 immunohistochemically.. Polyp samples showed an increased presence of cytokines in subepithelial connective tissue and a decreased appearance in epithelium when compared to controls. There were several very strong, strong, and moderate correlations among factors.. IL-6 strongly correlates with other cytokines as well as with the proliferation marker Ki-67, which suggests significant stimulation of this regulatory cytokine and its possible involvement in the pathogenesis of recurrent nasal polyps. IL-4, IL-7, IL-10, and IL-12 correlate with Ki-67, which suggests the possible involvement of these cytokines in tissue cell proliferation in the case of recurrent nasal polyps.

Topics: Cell Proliferation; Chronic Disease; Cytokines; Humans; Interleukin-1; Interleukin-10; Interleukin-12; Interleukin-4; Interleukin-6; Interleukin-7; Interleukin-8; Ki-67 Antigen; Nasal Polyps; Rhinitis; Sinusitis

We recently developed a ciprofloxacin and azithromycin sinus stent (CASS) to target recalcitrant infections in chronic rhinosinusitis (CRS). The objective of this study was to evaluate the anti-inflammatory activity of azithromycin released from the CASS and assess the impact on the integrity and function of primary human sinonasal epithelial cells (HSNECs).. Pseudomonas aeruginosa lipopolysaccharide (LPS)-stimulated HSNECs were treated with azithromycin and/or ciprofloxacin at concentrations attainable from CASS release. Interleukin-8 (IL-8) secretion was quantified by enzyme-linked immunosorbent assay (ELISA). Epithelial integrity (transepithelial resistance [TEER], paracellular permeability [fluorescein isothiocyanate-labeled dextran], lactate dehydrogenase [LDH] assays) and function (ciliary beat frequency [CBF]) were also evaluated.. Azithromycin significantly reduced secreted IL-8 from P. aeruginosa LPS-stimulated HSNECs at all concentrations tested (mean ± standard deviation; control = 5.77 ± 0.39 ng/mL, azithromycin [6 μg/mL] = 4.58 ± 0.40 ng/mL, azithromycin [60 µg/mL] = 4.31 ± 0.06, azithromycin [180 µg/mL] = 4.27 ± 0.26 ng/mL, p < 0.05). Co-incubation with azithromycin (6 µg/mL) and ciprofloxacin (2.4 µg/mL) in LPS-stimulated HSNECs also displayed a significant reduction in secreted IL-8 when compared to P. aeruginosa LPS alone (co-treatment = 4.61 ± 0.29 ng/mL, P. aeruginosa LPS = 7.35 ± 0.89 ng/mL, p < 0.01). The drugs did not negatively impact TEER, paracellular permeability, LDH release, or CBF, indicating retention of cell integrity and function.. Azithromycin decreased P. aeruginosa LPS IL-8 production in HSNECs at drug concentrations attainable with sustained release of azithromycin from the CASS. In addition to antibacterial activity, anti-inflammatory properties of the CASS should provide further benefit for patients with recalcitrant CRS.

Topics: Azithromycin; Ciprofloxacin; Epithelial Cells; Humans; Interleukin-8; Paranasal Sinuses; Sinusitis

Staphylococcus aureus colonization and release of enterotoxin B (SEB) has been associated with severe chronic rhinosinusitis with nasal polyps (CRSwNP). The pathogenic mechanism of SEB on epithelial barriers, however, is largely unexplored.. We investigated the effect of SEB on nasal epithelial barrier function.. SEB was apically administered to air-liquid interface (ALI) cultures of primary polyp and nasal epithelial cells of CRSwNP patients and healthy controls, respectively. Epithelial cell integrity and tight junction expression were evaluated. The involvement of Toll-like receptor 2 (TLR2) activation was studied in vitro with TLR2 monoclonal antibodies and in vivo in tlr2. SEB applied to ALI cultures of polyp epithelial cells decreased epithelial cell integrity by diminishing occludin and zonula occludens (ZO)-1 protein expression. Antagonizing TLR2 prevented SEB-induced barrier disruption. SEB applied in the nose of control mice increased mucosal permeability and decreased mRNA expression of occludin and ZO-1, whereas mucosal integrity and tight junction expression remained unaltered in tlr2. SEB damages nasal polyp epithelial cell integrity by triggering TLR2 in CRSwNP. Our results suggest that SEB might represent a driving factor of disease exacerbation, rather than a causal factor for epithelial defects in CRSwNP. Interfering with TLR2 triggering might provide a way to avoid the pathophysiological consequences of S. aureus on inflammation in CRSwNP.

Topics: Adolescent; Adult; Aged; Animals; Case-Control Studies; Cell Line; Enterotoxins; Female; Humans; In Vitro Techniques; Interleukin-6; Interleukin-8; Male; Mice; Mice, Knockout; Middle Aged; Nasal Mucosa; Nasal Polyps; Occludin; Permeability; Primary Cell Culture; Rhinitis; RNA, Messenger; Sinusitis; Staphylococcus aureus; Tight Junctions; Toll-Like Receptor 2; Young Adult; Zonula Occludens-1 Protein

IL-8 is an important chemokine implicated in the pathogenesis of chronic rhinosinusitis (CRS), but little is known about epigenetic regulation of IL8 in the pathogenesis of CRS.. We sought to investigate the relationship between the DNA methylation level in the IL8 proximal promoter and CRS in Han Chinese subjects.. Patients with chronic rhinosinusitis with nasal polyps (CRSwNP; n = 187), patients with chronic rhinosinusitis without nasal polyps (CRSsNP; n = 89), and control subjects (n = 57) were enrolled in 2 independent cohorts. Purified human nasal epithelial cells from each participant were assessed for percentage DNA methylation of CpG sites in the IL8 proximal promoter by using bisulfite pyrosequencing and for functional aspects of methylation status by using in vitro assays.. DNA methylation of CpG sites 1, 2, and 3, respectively, in the IL8 proximal promoter was significantly decreased in human nasal epithelial cells of patients with CRSwNP compared with that in patients with CRSsNP (P < .001) and control subjects (P < .001). Percentage of DNA methylation of the CpG3 site was correlated negatively with both tissue eosinophilic cationic protein (P < .01) and myeloperoxidase (P < .05) levels. IL-1β (P < .001) and TNF-α (P < .01) significantly increased IL8 expression accompanied by a reduction in methylation at the CpG3 site (P < .001). Electrophoretic mobility shift assays demonstrated that methylation status of CpG3 changed the binding of octamer-binding transcription factor 1 and nuclear factor κB.. Decreased DNA methylation of particularly CpG sites in the IL8 proximal promoter might play a role in the pathogenesis of CRSwNP.

Topics: Adolescent; Adult; Aged; Asian People; Chronic Disease; Cohort Studies; CpG Islands; DNA Methylation; Female; Humans; Interleukin-8; Male; Middle Aged; Nasal Polyps; Promoter Regions, Genetic; Respiratory Mucosa; Rhinitis; Sinusitis; Young Adult

High-mobility group box 1 (HMGB1) mediates various functions according to the location. We tried to investigate the role of HMGB1 in upper airway under hypoxic conditions. We cultured primary normal human nasal epithelium (NHNE) cells under hypoxic conditions and evaluated the movement of HMGB1 by western blotting, immunofluorescence, and enzyme-linked immunosorbent assay (ELISA). Reactive oxygen species (ROS) level was evaluated to estimate the translocation mechanism of HMGB1. The role of secreted HMGB1 was evaluated by ELISA assay. Furthermore, we collected human nasal mucosa samples and nasal lavage fluids from patients conditioned under hypoxic and non-hypoxic environment, and compared the expression of HMGB1 in human nasal mucosa samples by immunohistochemistry and the levels of HMGB1 in lavage fluids using ELISA assay. Hypoxia induced translocation of HMGB1 into the extracellular area and it was dependent on ROS produced by dual oxidase 2. Secreted HMGB1 was involved in the upregulation of interleukin (IL)-8. In human samples, HMGB1 was translocated from nucleus to the cytoplasm in hypoxic-conditioned nasal mucosa. HMGB1 was increased in nasal lavage samples of chronic rhinosinusitis patients, whose sinus mucosa was supposed to be hypoxic as compared with controls. We suggest that HMGB1 is secreted in hypoxic condition via ROS-dependent mechanism and secreted HMGB1 participates in IL-8 upregulation mediating inflammatory response.

Topics: Adult; Cells, Cultured; Chronic Disease; Dual Oxidases; Female; HMGB1 Protein; Humans; Hypoxia; Interleukin-8; Male; Middle Aged; Nasal Mucosa; Reactive Oxygen Species; Respiratory Mucosa; Rhinitis; Sinusitis; Up-Regulation; Young Adult

Topics: Chronic Disease; Humans; Immunity, Innate; Interleukin-33; Interleukin-8; Rhinitis; Sinusitis

Fibroblasts are major supporting cells in nasal mucosa and can induce inflammatory process with recruitment of inflammatory cells. Airborne fungi have been suggested as an etiologic factor of chronic rhinosinusitis (CRS). The aim of this study was to investigate the interaction between airborne fungi and pattern recognition receptors (PRRs) in nasal fibroblasts.. Primary nasal polyp fibroblasts were cultured with Alternaria and Aspergillus for 48h. To determine the production of chemical mediators interleukine-6 (IL-6), IL-8, granulocyte-macrophage colony stimulating factor (GM-CSF), eotaxin, and regulated on activation normal T expressed and secreted (RANTES) were measured with enzyme immunoassay methods. PRRs for toll-like receptors (TLRs) and protease-activated receptors (PARs) mRNA were determined with reverse transcription polymerase chain reaction (RT-PCR). To determine the role of PRRs, fibroblasts were treated with small interfering RNA (siRNA).. IL-6 and IL-8 productions were significantly increased by 50 and 100μg/ml of Alternaria. However, GM-CSF, eotaxin, and RANTES productions did not change. Aspergillus did not influence the production of chemical mediators from nasal polyp fibroblasts. TLR2 and TLR5 mRNA expressions were significantly increased by fungi and these two TLRs were associated with the production of IL-6 and IL-8.. Alternaria interacts as a pathogen-associated molecular pattern with the PRRs, such as TLR2 and TLR5, which induce the production of inflammatory chemical mediators from nasal polyp fibroblasts. Airborne fungi enhance the innate immune defense mechanism and may be associated with the pathogenesis of nasal inflammatory diseases.

Topics: Adult; Alternaria; Aspergillus; Cells, Cultured; Chemokine CCL5; Chemokines; Chronic Disease; Female; Fibroblasts; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Interleukin-6; Interleukin-8; Male; Middle Aged; Mycoses; Nasal Polyps; Receptors, Pattern Recognition; Receptors, Proteinase-Activated; Reverse Transcriptase Polymerase Chain Reaction; Rhinitis; RNA, Messenger; RNA, Small Interfering; Sinusitis; Toll-Like Receptors

T-helper 2 (Th2) inflammation is a hallmark of chronic rhinosinusitis with nasal polyps (CRSwNP) although the pathogenesis is poorly understood. P-glycoprotein (permeability glycoprotein, P-gp) is an efflux pump that is capable of regulating cytokine transport and is expressed within sinonasal mucosa. The purpose of this study was to examine if the oversecretion of interleukin 5 (IL-5) and thymic stromal lymphopoietin (TSLP) in CRSwNP could be explained through P-gp-mediated secretory pathways.. Fifteen ethmoid mucosal explants were harvested from patients with CRS (n = 10) and CRSwNP (n = 10) and stimulated with Staphylococcus aureus enterotoxin B (SEB). P-gp was inhibited using zosuquidar trihydrochloride (herein Zosuquidar). P-gp expression was measured using real-time polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA). IL-5, IL-8, and TSLP secretion were quantified using ELISA.. P-gp protein was overexpressed in CRSwNP (28.32 ± 25.94 ng/mL per mg explant) as compared to CRS (10.74 ± 8.61; p = 0.01, 2-tailed Mann-Whitney U test). There was no difference in messenger RNA (mRNA) expression. SEB induced a significant increase in IL-5 and TSLP but not IL-8 secretion relative to control in the CRSwNP explants only. Subsequent P-gp inhibition significantly reduced IL-5 and TSLP secretion (p = 0.04 for both, 2-tailed Student t test) to control levels. The concentration of IL-5 and TSLP secretion were strongly and significantly correlated to the concentration of P-gp within the same explant (IL-5: r = 0.791, p = 0.001; TSLP: r = 0.687, p = 0.003; 2-tailed Spearman's rank-order correlation).. P-gp protein is expressed at higher concentrations in CRSwNP as compared to CRS. This overexpression directly contributes to the relative hypersecretion of IL-5 and TSLP. These findings suggest a novel mechanism for Th2 skewing in CRSwNP.

Topics: ATP Binding Cassette Transporter, Subfamily B, Member 1; Cells, Cultured; Chronic Disease; Cytokines; Dibenzocycloheptenes; Enterotoxins; Humans; Interleukin-5; Interleukin-8; Nasal Mucosa; Organ Culture Techniques; Quinolines; Rhinitis; Sinusitis; Staphylococcus aureus; Thymic Stromal Lymphopoietin

Alarmins play important roles in the pathogenesis of inflammatory and autoimmune diseases. However, the role of the alarmin protein high-mobility group box 1 (HMGB1) in upper airway inflammation is unclear.. To determine if HMGB1 is present in the nasal mucosa and, if so, to elucidate its role in upper airway inflammation.. Nasal secretions were collected from a total of 32 patients with chronic rhinosinusitis with nasal polyp, allergic rhinitis, and control subjects. The concentration of HMGB1 in nasal secretions and its tissue and cellular localization were examined by enzyme immunoassays and immunofluorescent staining of nasal polyps and cultured nasal epithelial cells. We then examined whether nasal epithelial cells secrete HMGB1 after inflammatory stimulation by tumor necrosis factor (TNF) α. The effects of HMGB1 on the production and secretion of interleukin (IL) 6 and IL-8 were also examined in cultured nasal epithelial cells.. Significantly higher concentrations of HMGB1 were found in nasal secretions from patients with chronic rhinosinusitis with nasal polyp or allergic rhinitis compared with the control subjects. HMGB1 expression was localized in the nuclei of epithelial cells and other constitutive cells in nasal polyps and in the nuclei of cultured nasal epithelial cells. TNF-α stimulated the production and secretion of HMGB1 by cultured nasal epithelial cells. HMGB1 stimulated the production and secretion of IL-6 and IL-8 by cultured nasal epithelial cells, and anti-toll-like receptor 4 blocking antibody significantly inhibited HMGB1-induced secretion of IL-6 and IL-8.. Nasal secretions contain substantial amounts of HMGB1. TNF-α stimulates the production of HMGB1, which, in turn, upregulates the production and secretion of IL-6 and IL-8 by nasal epithelial cells via toll-like receptor 4, which indicated that HMGB1 plays an important role in the pathogenesis of upper airway inflammation.

Topics: Adult; Aged; Antibodies, Blocking; Cells, Cultured; Chronic Disease; Epithelial Cells; Female; HMGB1 Protein; Humans; Interleukin-6; Interleukin-8; Male; Middle Aged; Nasal Mucosa; Nasal Polyps; Rhinitis, Allergic; Signal Transduction; Sinusitis; Toll-Like Receptor 4; Tumor Necrosis Factor-alpha

Chronic rhinosinusitis without nasal polyps (CRSsNP) is a common chronic disease and the etiology remains unclear. Thromboxane A2 (TXA2) participates in platelet aggregation and tissue inflammation. In this study, the CXCL1/8 chemokine and TXA2-TP receptor expression in the CRSsNP mucosa was investigated.. Immunohistochemistry, chemokine release assay by ELISA, RT-PCR, Real-time PCR, Western blotting, pharmacological and siRNA knockdown analysis were applied in the CRSsNP tissue specimen and cultured nasal mucosa-derived fibroblasts.. The immunohistochemistry results indicated that CXCL1 and CXCL8 were highly expressed in the CRSsNP mucosa compared with the controls; however, the TP receptors were expressed in both mucosa. Therefore, U46619 and IBOP, a TXA2 analog and TP agonist, were used to explore the role of TP activation in CXCL1/8 expression; both of these induced CXCL1/8 mRNA and protein expression in CRSsNP mucosa-derived fibroblasts. U46619 phosphorylated PI-3K, cyclic AMP (cAMP)/PKA, PKC, and cAMP response element (CREB). Activation of cAMP/PKA, PKC, and CREB was the major pathway for cxcl1/8 gene transcription. Pharmacological and siRNA knockdown analyses revealed that activation of cAMP/PKA and PKCμ/PKD pathways were required for CREB phosphorylation and PKA/C crosstalked with the PI-3K pathway.. Our study provides the first evidence for abundant TP receptor and CXCL1/8 expression in human CRSsNP mucosa and for TXA2 stimulation inducing CXCL1/8 expression in nasal fibroblasts primarily through TP receptor, cAMP/PKA, PKCμ/PKD, and CREB-related pathways.

Topics: 15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid; Bridged Bicyclo Compounds, Heterocyclic; Case-Control Studies; Cells, Cultured; Chemokine CXCL1; Cyclic AMP Response Element-Binding Protein; Fatty Acids, Unsaturated; Fibroblasts; Interleukin-8; Nasal Mucosa; Phosphatidylinositol 3-Kinases; Protein Kinase C; Receptors, Thromboxane A2, Prostaglandin H2; Rhinitis; Second Messenger Systems; Sinusitis; Thromboxane A2

Nasal secretions include cytokines and inflammatory mediators that are involved in the pathogenesis of upper airway inflammation.. We tried to find unknown biomolecules that are involved in the pathogenesis of chronic rhinosinusitis (CRS).. We collected nasal mucosal secretions from patients who were diagnosed as having CRS and who underwent endoscopic sinus surgery. A total of 63 patients who underwent nasal secretion collection were reviewed. Enzyme-linked immunosorbent assay was performed by using nasal lavage samples to evaluate which biomolecules were associated with the severity of inflammation based on the Lund-Mackay score. By using human nasal epithelial cells, we performed Western blot, real-time polymerase chain reaction, and enzyme-linked immunosorbent assay to evaluate the secretory mechanism of heat shock protein (HSP) 70.. We found that the level of interleukin 8 and HSP70 were significantly associated with the Lund-Mackay score and interleukin 17C, C-X-C motif chemokine 10, and HSP27 were not significantly associated. HSP70 was also significantly associated with the surgical outcome of the enrolled patients. Furthermore, we found that exposure to hypoxia and treatment of lipoteichoic acid induced the secretion of HSP70 but that lipopolysaccharide did not induce the secretion of HSP70 in human nasal epithelial cells.. Our findings indicated that HSP70 might play a role in the pathogenesis of CRS and the possibility of HSP70 as a biomolecule that represents the severity of CRS.

Topics: Adolescent; Adult; Aged; Cells, Cultured; Chronic Disease; Female; HSP70 Heat-Shock Proteins; Humans; Interleukin-8; Lipopolysaccharides; Male; Middle Aged; Rhinitis; Severity of Illness Index; Sinusitis; Teichoic Acids

Chronic rhinosinusitis (CRS) is an inflammatory disorder of the nose and paranasal sinuses. Staphylococcus aureus is increasingly linked with CRS exacerbations. Little is known about how bacteria activate inflammatory pathways that contribute to CRS.. To develop an in vitro coculture system to explore how infection with S aureus stimulates innate immune responses of sinonasal epithelial cells (SNECs).. Sinonasal epithelial cells were collected from 13 patients during endoscopic sinus surgery and grown in culture at the air-liquid interface from July 2014 through December 2014.. Differentiated SNECs from control individuals, patients with CRS with nasal polyps (CRSwNPs), and patients with CRS without nasal polyps (CRSsNPs) were infected with S aureus at 3 different concentrations for 24 hours.. Growth of S aureus and viability of SNECs were measured. Expression of inflammatory markers and innate immune genes was measured by reverse transcription-polymerase chain reaction. Basal secretion of interleukin 8 was determined by enzyme-linked immunosorbent assay.. Cultured SNECs from patients with CRSsNPs demonstrated a significant increase (P < .05) in expression of interleukin 8 (23-fold to 82-fold) and tumor necrosis factor (11-fold to 61-fold) at all the tested concentrations of S aureus. Control or CRSwNP SNECs demonstrated a significant increase (P < .05) in expression of interleukin 8 (47-fold and 50-fold, respectively) and tumor necrosis factor (106-fold and 58-fold, respectively) at the higher inoculum of S aureus. Basal secretion of inflammatory markers correlated with expression changes. No significant changes in expression were observed for the helper T cell, subtype 2, inflammatory mediators tested.. In this study, we developed a model to study early innate immune-mediated changes in SNECs cocultured at an air-liquid interface with bacteria. We also demonstrated that bacterial burden can be detected by SNECs in the absence of adaptive immune-mediated responses. The CRSsNP SNECs are more sensitive to S aureus burden than control or CRSwNP SNECs. Future studies will further develop this infection model and explore the SNEC innate immune response to bacteria.

Topics: Bacterial Load; Case-Control Studies; Cell Survival; Chronic Disease; Coculture Techniques; Epithelial Cells; Humans; Immunity, Innate; Interleukin-8; Nasal Mucosa; Nasal Polyps; Rhinitis; Sinusitis; Staphylococcus aureus; Tumor Necrosis Factor-alpha

High mobility group box 1 (HMGB1) protein is a chromatin protein that functions as a proinflammatory cytokine when secreted in response to inflammatory stimuli. The purpose of this study was to determine the relationship between the HMGB1 level in nasal secretions and the severity of inflammation in chronic rhinosinusitis.. This was a cross-sectional study.. Nasal secretions were obtained by irrigation of the affected sinonasal cavities with normal saline. Total 63 nasal lavage fluid samples were collected from 38 patients with chronic rhinosinusitis who underwent endoscopic sinus surgery. Levels of HMGB1 and tumor necrosis factor alpha, interleukin (IL)-1β, and IL-8 were determined by enzyme-linked immunoassay. Severity of inflammation was assessed by the Lund-Mackay scoring system, which is based on preoperative computed tomography scans. Concurrent medical disorders, presence of nasal polyps, septal deviation, and allergic rhinitis were also investigated.. The level of HMGB1 in nasal lavage fluid was positively correlated with the Lund-Mackay score. The score was the only factor associated with HMGB1 by univariate and multivariate analysis. Other cytokines, with the exception of IL-8, were not correlated with the Lund-Mackay score.. Our results showed that HMGB1 is secreted into the extracellular area space in the upper airway, and HMGB1 levels in nasal lavage fluid correlate with severity of inflammation, as assessed by the Lund-Mackay staging system for chronic rhinosinusitis. These results provide evidence for HMGB1 as an inflammatory mediator associated with the severity of chronic rhinosinusitis.

Topics: Adolescent; Adult; Aged; Chronic Disease; Cross-Sectional Studies; Enzyme-Linked Immunosorbent Assay; Female; HMGB1 Protein; Humans; Immunohistochemistry; Inflammation; Interleukin-1beta; Interleukin-8; Male; Middle Aged; Nasal Lavage Fluid; Rhinitis; Severity of Illness Index; Sinusitis; Tumor Necrosis Factor-alpha; Young Adult

Chronic rhinosinusitis (CRS) is a chronic inflammatory disease of the sinonasal mucosa either accompanied by polyp formation (CRSwNP) or without polyps (CRSsNP). CRSsNP accounts for the majority of CRS cases and is characterized by fibrosis and neutrophilic inflammation. However, the pathogenesis of CRS, especially CRSsNP, remains unclear. Immunohistochemistry of CRSsNP specimens in the present study showed that the submucosa, perivascular areas, and the mucous glands were abundant in fibroblasts. Therefore, we investigated the effects bradykinin (BK), an autacoid known to participate in inflammation, on human CRSsNP nasal mucosa-derived fibroblasts (NMDFs). BK increased CXCL1 and -8 secretion and mRNA expression with EC50 ranging from 0.15~0.35 μM. Moreover, BK enhanced cell proliferation and upregulated the expressions of proinflammatory molecules, including cell adhesion molecules (CAMs) and cyclooxygenase (COX)-1 and -2. These functionally caused an increase in monocyte adhesion to fibroblast monolayer. Using pharmacological intervention and BKR siRNA knockdown, we demonstrated that the BK-induced CXCL chemokine release, cell proliferation and COX and CAM expressions were mainly through the B2 receptor (B2R). Accordingly, the B2R was preferentially expressed in the NMDFs than B1R. The B2R was highly expressed in the CRSsNP than the control specimens, while the B1R and kininogen (KNG)/BK expression slightly increased in the CRSsNP mucosa. Collectively, we report here for the first time that fibroblasts, KNG/BK, and BKRs are overexpressed in CRSsNP mucosa and BK upregulates chemokine expression, proliferation, and proinflammatory molecule expression in NMDFs via B2R activation, which lead to a functional increase in monocyte-fibroblast interaction. Our findings reveal a critical role of fibroblast, KNG/BK, and BKRs in the development of CRSsNP.

Topics: Bradykinin; Cell Adhesion; Cell Proliferation; Cells, Cultured; Chemokine CXCL1; Chronic Disease; Fibroblasts; Gene Expression; Humans; Interleukin-8; Nasal Mucosa; Receptor, Bradykinin B1; Receptor, Bradykinin B2; Rhinitis; Sinusitis

Firefighters exposed to World Trade Center (WTC) dust have developed chronic rhinosinusitis (CRS) and abnormal forced expiratory volume in 1 s (FEV1). Overlapping but distinct immune responses may be responsible for the clinical manifestations of upper and lower airway injury. We investigated whether a panel of inflammatory cytokines, either associated or not associated with WTC-LI, can predict future chronic rhinosinusitis disease and its severity.. Serum obtained within six months of 9/11/2001 from 179 WTC exposed firefighters presenting for subspecialty evaluation prior to 3/2008 was assayed for 39 cytokines. The main outcomes were medically managed CRS (N = 62) and more severe CRS cases requiring sinus surgery (N = 14). We tested biomarker-CRS severity association using ordinal logistic regression analysis.. Increasing serum IL-6, IL-8, GRO and neutrophil concentration reduced the risk of CRS progression. Conversely, increasing TNF-α increased the risk of progression. In a multivariable model adjusted for exposure intensity, increasing IL-6, TNF-α and neutrophil concentration remained significant predictors of progression. Elevated IL-6 levels and neutrophil counts also reduced the risk of abnormal FEV1 but in contrast to CRS, increased TNF-α did not increase the risk of abnormal FEV1.. Our study demonstrates both independent and overlapping biomarker associations with upper and lower respiratory injury, and suggests that the innate immune response may play a protective role against CRS and abnormal lung function in those with WTC exposure.

Topics: Biomarkers; Dust; Firefighters; Forced Expiratory Volume; Humans; Inflammation; Interleukin-6; Interleukin-8; Lung Injury; Male; Neutrophils; New York; Occupational Exposure; Respiratory Tract Diseases; Rhinitis; Risk Assessment; Risk Factors; September 11 Terrorist Attacks; Severity of Illness Index; Sinusitis; Tumor Necrosis Factor-alpha

Asian dust storms (ADS) contain various airborne particles that may augment airway inflammation by increasing the level of interleukin-8. The objective of the study was to investigate the association of exposure to an ADS with worsening of symptoms of adult asthma and the effect of ADS particles on interleukin-8 transcriptional activity.. The subjects were 112 patients with mild to moderate asthma who recorded scores for their daily upper and lower respiratory tract symptoms and measured morning peak expiratory flow (PEF) from March to May 2011. Interleukin-8 transcriptional activity was assessed in THP-G8 cells that were exposed to airborne particles collected during days of ADS exposure.. Of the 112 patients, 31 had comorbid allergic rhinitis (AR) and/or chronic sinusitis (CS), and had worsened scores for upper respiratory tract symptoms on ADS days compared to non-ADS days. Scores for lower respiratory tract symptoms during ADS days were higher than non-ADS days in all patients. Three patients also had unscheduled hospital visits for exacerbation of asthma on ADS days. However, there was no significant difference in daily morning PEF between ADS and non-ADS days. Airborne particles collected on ADS days induced interleukin-8 transcriptional activity in THP-G8 cells compared to the original soil of the ADS.. Exposure to an ADS aggravates upper and lower tract respiratory symptoms in patients with adult asthma. ADS airborne particles may increase airway inflammation through enhancement of interleukin-8 transcriptional activity.

Topics: Aged; Air Pollutants; Asthma; Dust; Enzyme-Linked Immunosorbent Assay; Female; Humans; Interleukin-8; Japan; Luciferases; Male; Middle Aged; Particulate Matter; Peak Expiratory Flow Rate; Rhinitis, Allergic; Rhinitis, Allergic, Perennial; Sinusitis; Wind

The primary human sinonasal epithelial cell culture (HSNEC) allows for in-vitro modelling of mucosal responses to topical therapy. Cultures grown from healthy donors may underestimate changes in individuals with chronic sinonasal disease thereby yielding inaccurate results with respect to this large patient population. The purpose of this study was to analyse HSNECs derived from patients with chronic rhinosinusitis with nasal polyposis (CRSwNP) to determine whether expected disease dependent variables salient to topical drug delivery persist in culture.. Cultures were grown from patients with CRSwNP. Ciliary beat frequency (CBF) (basal and stimulated), permeability (trans and paracellular), inflammatory response, and glucocorticoid dose response were measured and compared with healthy controls.. Methylcholine stimulated CBF was greater in CRSwNP versus controls (ΔCBF(60 min) 7.25 ± 1.02 vs 0.89 ± 1.04 Hz, respectively). Paracellular permeability was greater in CRSwNP versus controls (basolateral dextran(120 min) 18.97 ± 3.90 vs 11.31 ± 4.35 µg/ml, respectively). Lipopolysaccharide (0.1 mg/ml) stimulated interleukin-6 (IL-6) and IL-8 secretion was increased in CRSwNP versus controls (IL-6 Δbaseline 1738.72 ± 654.82 vs 1461.61 ± 533.51%, respectively; IL-8 Δbaseline 137.11 ± 0.83 vs 111.27 ± 0.67%, respectively). CRSwNP cultures were more sensitive than controls to dexamethasone (1 µg/ml) dependent IL-6 and IL-8 suppression.. HSNECs derived from patients with CRSwNP retained their primary phenotype with respect to ciliary function, epithelial permeability, irritant induced inflammatory cytokine secretion, and glucocorticoid dose response.

Topics: Administration, Topical; Case-Control Studies; Cells, Cultured; Chronic Disease; Cilia; Dexamethasone; Dose-Response Relationship, Drug; Epithelial Cells; Glucocorticoids; Humans; Interleukin-6; Interleukin-8; Models, Biological; Nasal Mucosa; Nasal Polyps; Phenotype; Rhinitis; Sinusitis

The examination was applied to 81 children aged 5-15 years, including 64 children with diagnosis of rhinosinusitis. The control group consisted of 17 healthy children. The set of laboratory clinical diagnostic techniques was applied to detect the causes of pathology. It is established that children with rhinosinusitis suffered from concurrent bacterial and virus infections. The morphologic presentation of mucous membrane of nasal cavity reflects the etiologic factor and the stage of inflammatory process. The detection of concentration of IL-4, IL-6 and IL-8 of blood serum gives a possibility to diagnose children with combined mechanisms of development of rhinosinusitis.

Topics: Adolescent; Child; Child, Preschool; Enzyme-Linked Immunosorbent Assay; Humans; Interleukin-4; Interleukin-6; Interleukin-8; Nasal Cavity; Rhinitis; Sinusitis

To investigate the expressions of LL-37 and IL-8 in chronic sinusitis with nasal polyps.. Semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemical staining were used to detect the expressions of LL-37 and IL-8 in nasal polyp tissues of 31 patients with chronic sinusitis and inferior turbinate tissues of 11 patients with chronic rhinitis.. LL-37 and IL-8 mRNA were all positively expressed in all nasal polyps and inferior turbinate tissues. There were significant increases of LL-37 and IL-8 mRNA expressions in nasal polyps compared with the inferior turbinate tissues (P < 0.01). There were also significant increases of positive expression rates of LL-37 and IL-8 protein in nasal polyps, compared with the inferior turbinate tissues (P < 0.01). There was a positive relationship between the mRNA and protein expressions of LL37 and IL-8 (P < 0.01).. The expressions of LL-37 and IL-8 in nasal polyps suggest that they may play a role in the pathogenesis of chronic sinusitis. Besides its innate immune, LL-37 could enhance human body's anti-infected function by increasing acquired immune.

Topics: Adult; Antimicrobial Cationic Peptides; Cathelicidins; Chronic Disease; Female; Humans; Interleukin-8; Male; Middle Aged; Nasal Mucosa; Nasal Polyps; Sinusitis; Young Adult

The aim of this study was to elucidate the role of IL-8 for neutrophil recruitment in nonallergic CRS patients.. After coculture of Streptococcus pneumoniae (SP) with the mucosal epithelial cells (MECs) from non-CRS patients, at three different SP/MEC (1/1, 10/1, 100/1) ratios, the expression of IL-8 mRNA and the concentration of IL-8 were measured by RT-PCR and ELISA. The expression of CD11b/CD18 on neutrophils and E-selectin/ICAM-1 on endothelial cells and the adherence between neutrophils and human umbilical vascular endothelial cells (HUVECs) were determined by flow cytometric analysis, ELISA, and RIA, respectively.. IL-8 concentration and IL-8 mRNA expression continued to increase from 3 hours after incubation in SP number-dependent manner. The expression of CD11b/CD18 on neutrophils and E-selectin/ICAM-1 on HUVECs, and the adherence between neutrophils and HUVECs were significantly increased in 10 SP/MEC-CM, and the increments were significantly blocked by anti-IL-8 antibody.. MEC and IL-8 are major factors for neutrophil recruitment in nonallergic CRS.

Topics: Adolescent; Adult; Animals; Bacterial Infections; CD11b Antigen; CD18 Antigens; Cells, Cultured; Chronic Disease; Epithelial Cells; Female; Humans; Interleukin-8; Male; Middle Aged; Nasal Mucosa; Neutrophil Infiltration; Rhinitis; Sinusitis; Young Adult

In patients with chronic fungal sinusitis, concentrations of interleukin-8 (IL-8), immunoglobulin E (IgE), and soluble intercellular adhesion molecule-1 (sICAM-1) were compared in paranasal sinus aspirates and serum. Furthermore, immunological effects of macrolide treatment of our patients with chronic fungal rhinosinusitis were also studied.. In our cohort study, 108 patients with chronic rhinosinusitis undergoing sinus surgery were selected. Sinus aspirates were collected, and used for immunological assasy and cultured for fungal study. All patients were examined for the presence of characteristic allergic mucin of chronic allergic fungal rhinosinusitis and this was confirmed later by measurement of total serum IgE.. Our cases were classified into 3 groups: chronic rhinosinusitis with positive fungal culture and negative allergic mucin, chronic rhinosinusitis with positive fungal culture and positive allergic mucin and chronic rhinosinusitis without fungal growth. A control group was included. We found 57.4% of the patient cultures positive for fungus and 36.4% of the control subjects. Aspergillus ssp. were the most prevalent followed by Bipolaris ssp., and Curvularia. IgE levels were increased in group II compared to group I, III and IV. ICAM-1 and IL-8 levels were increased in groups I, II and III compared to the control group. Erythromycin given in group II decreased the levels of IL-8 and ICAM-1.. Aspergillus species were the most common. These results confirm the role of ICAM-1 and IL-8 in all types of rhinosinusitis. Erythromycin modulated the immune status of the patients.

Topics: Adult; Anti-Bacterial Agents; Chronic Disease; Erythromycin; Female; Humans; Immunoglobulin E; Intercellular Adhesion Molecule-1; Interleukin-8; Male; Paranasal Sinuses; Rhinitis; Sinusitis

To explore the relationship between the bacterial biofilm, allergy, inflammatory cytokines and refractory rhinosinusitis.. According to the assessment of symptom by visual analogue scale and nasal endoscopy by Lund-Kennedy system, 19 patients with persistent chronic rhinosinusitis and 6 patients with curative chronic rhinosinusitis were recruited from 340 chronic rhinosinusitis patients underwent functional endoscopic sinus surgery for the first time and received standard medication postoperatively. These cases were divided into the study group and the control group respectively. Electron microscope scanning and enzyme linked immunosorbent assay were used to observe the bacterial biofilm formation on ethmoid sinus mucosal surface and detect the concentration of specific IgE and interleukin (IL) 4, IL-5, IL-6, and IL-8 in ethmoid sinus mucosal tissue. Then the differences between the two groups of cases were analyzed statistically.. (1) Bacterial biofilms: nineteen cases from the study group were all found biofilm formation in ethmoid sinus, besides the damage of epithelial cells and cilia. Six cases from the control group were found no biofilm, with the integrity of epithelial cells and cilia. Statistical analysis showed significant differences in biofilm detection rate between the two groups (P < 0.01). (2) Specific IgE: specific IgE was detected in nine cases from the study group and was not found in any case from the control group, with a significant difference between them (P < 0.05). (3) IL: IL concentration was all detected under normal value both in the study group and the control group, except for only 2 cases from the study group with higher IL-8 concentration. Positive reaction to IL was detected as follows: IL-4 (7 cases), IL-5 (14 cases), IL-6 (15 cases), and IL-8 (12 cases) in the study group; IL-4 (1 case), IL-5 (5 cases), IL-6 (5 cases), and IL-8 (5 cases) in control group. No difference in IL detection rate was found between the two groups (P > 0.05).. Bacterial biofilm and allergic inflammation are important factors resulting into refractory chronic rhinosinusitis.

Topics: Adult; Bacteria; Biofilms; Case-Control Studies; Chronic Disease; Female; Humans; Immunoglobulin E; Interleukin-4; Interleukin-5; Interleukin-6; Interleukin-8; Male; Middle Aged; Reference Values; Sinusitis; Young Adult

The impact of Staphylococcus aureus in the pathogenesis of chronic rhinosinusitis is not well understood. Therefore, we investigated primary human nasal epithelial cell cultures for their ability to produce IL-8, growth-related oncogene-alpha, and IL-6 via stimulation with trypsin and culture supernatants of different S. aureus strains and phenotypes. Inhibition of cytokine synthesis was performed using a glucocorticoid, a serine protease inhibitor, and a cysteine protease inhibitor. Finally, signal transduction pathways were analyzed by quantifying phosphorylated forms of MAPKs (PI3K, ERK, and p38) and DNA-binding assays that quantified NF-kappaB and its inhibition using BAY11-7085. In vitro studies showed that the induction of IL-8, growth-related oncogene-alpha, and IL-6 by S. aureus culture supernatants was significantly inhibited by the serine protease inhibitor. In contrast, steroids and the cysteine protease inhibitor had little effect. Activation of NF-kappaB was observed after cell treatment with trypsin and bacterial supernatants, and was inhibited by BAY11-7085 and the serine protease inhibitor. S. aureus serine proteases were identified to modulate chemokine synthesis and activate NF-kappaB in nasal epithelial cells, and may therefore be relevant for the pathophysiology of chronic rhinosinusitis.

Topics: Cells, Cultured; Chemokine CXCL1; Electrophoretic Mobility Shift Assay; Enzyme-Linked Immunosorbent Assay; Epithelial Cells; Humans; Immunomodulation; Interleukin-6; Interleukin-8; Metalloendopeptidases; Middle Aged; Nasal Mucosa; NF-kappa B; Rhinitis; Serine Proteases; Signal Transduction; Sinusitis; Staphylococcal Infections; Staphylococcus aureus

Chronic rhinosinusitis (CRS) is characterized by persistent mucosal inflammation and frequent exacerbations.. To determine whether innate epithelial responses to cigarette smoke or bacterial or viral pathogens may be abnormal in CRS leading to an inappropriate inflammatory response.. Primary nasal epithelial cells (PNECs) were grown from middle turbinate biopsies of 9 healthy controls and 11 patients with CRS. After reaching 80% to 90% confluence, PNECs were exposed to medium or cigarette smoke extract (CSE) 5% (vol/vol) for 1 hour, washed, then stimulated with staphylococcal lipoteichoic acid, LPS, or double-stranded RNA (dsRNA). After 24 hours, gene expression was quantified by QRT-PCR.. At baseline, PNECs revealed elevated TNF-alpha and growth-related oncogene-alpha (a C-X-C chemokine)/CXCL1 (GRO-alpha) (4-fold increase, P = .02; and 16-fold increase, P = .004, respectively) in subjects with CRS compared with controls with normal levels of IL-1beta, IL-6, IL-8/CXCL8, human beta-defensin-2, monocyte chemoattractant protein 2/CCL8, monocyte chemoattractant protein 3/CCL7, and regulated upon activation, normal T-cell expressed and secreted (RANTES)/CCL5. Immunostaining of nasal biopsies, however, revealed comparable epithelial staining for TNF-alpha, GRO-alpha, and RANTES. There were no differences in mRNA induction by CSE, TNF-alpha, lipoteichoic acid, LPS, or dsRNA alone. The combination of CSE+dsRNA induced exaggerated RANTES (12,115-fold vs 1500-fold; P = .03) and human beta-defensin-2 (1120-fold vs 12.5-fold; P = .05) in subjects with CRS. No other genes were differentially induced. Furthermore, CSE+dsRNA induced normal levels of IFN-beta, IFN-lambda1, and IFN-lambda2/3 mRNA in subjects with CRS.. Cigarette smoke extract plus dsRNA induces exaggerated epithelial RANTES expression in patients with CRS. We propose that an analogous response to cigarette smoke plus viral infection may contribute to acute exacerbations and eosinophilic mucosal inflammation in CRS.

Topics: Adult; Aged; beta-Defensins; Cells, Cultured; Chemokine CCL5; Chemokine CCL7; Chronic Disease; Complex Mixtures; Epithelial Cells; Female; Gene Expression Regulation; Humans; Immunochemistry; Interferon-beta; Interferons; Interleukin-1beta; Interleukin-6; Interleukin-8; Interleukins; Lipopolysaccharides; Male; Middle Aged; Nasal Mucosa; Rhinitis; RNA, Double-Stranded; RNA, Messenger; Sinusitis; Tobacco Smoke Pollution; Toll-Like Receptor 3; Tumor Necrosis Factor-alpha

Bacterial etiology of chronic rhinosinusitis (CRS) still remains controversial. Whereas Staphylococcus aureus enterotoxins have been detected in CRS, the impact of Staphylococcus epidermidis, a major commensal inhabitant of the nose, has not been studied. Among others, serine and cysteine proteases have been identified as factors of virulence in S. epidermidis.. S. epidermidis was examined in tissue biopsies of 30 CRS patients (16 with nasal polyposis) using standard procedures. Primary human nasal epithelial cells from inferior nasal turbinates (HNECs), from nasal polyps (NPECs) and A549 airway epithelial cells were stimulated with S. epidermidis supernatants DSM20044 or ATCC35984 and the IL-8 and GRO-alpha response was quantified by ELISA. Protease-triggered chemokine responses and involvement of NF-kappaB were investigated by addition of protease or NF-kappaB inhibitors. Activation of NF-kappaB was demonstrated by quantitative DNA binding assay.. S. epidermidis was the most frequently isolated bacteria in the majority of CRS patients. HNECs and NPECs revealed no different IL-6 and IL-8 synthesis following stimulation with DSM20044 or ATCC35984. Stimulation of HNECs and A549 cells with S. epidermidis supernatants resulted in increased IL-8 and GRO-alpha expression which could be suppressed by the serine protease inhibitor AEBSF and the NF-kappaB inhibitor BAY 11 but not by the cysteine protease inhibitor E64. Results obtained for A549 cells were similar to HNECs.. S. epidermidis was present in the majority of CRS specimens. Proinflammatory impact of S. epidermidis supernatants on nasal epithelial cells was demonstrated by serine protease-triggered and NF-kappaB-dependent chemokine responses.

Topics: Cell Line; Cells, Cultured; Chemokine CXCL1; Chronic Disease; Epithelial Cells; Humans; Interleukin-6; Interleukin-8; Nasal Mucosa; Nasal Polyps; NF-kappa B; Nitriles; Rhinitis; Serine Proteinase Inhibitors; Sinusitis; Staphylococcus epidermidis; Sulfones

The effects of protease-activated receptor-2 (PAR-2) stimulation on inflammation mechanisms of chronic rhinosinusitis (CRS) are still unknown.. PAR-2 receptor expression was investigated by immunohistochemistry and Taqman mRNA analysis in the mucosa of different rhinosinusitis entities. In primary nasal epithelial cell cultures, the function of PAR-2 and its ability to produce CXC, CC chemokines, and IL-6 were measured by calcium mobilization and stimulation tests. Inhibition tests were performed using cortisone, serine protease inhibitors, cysteine protease inhibitors, Pertussis toxin (PTX) and nuclear transcription factor (NF-kappaB) inhibition (BAY 11-7085). Signal transduction pathways were analysed by electromobility shift assays (EMSA) and NF-kappaB binding studies.. The expression of PAR-2 was found to be increased in CRS specimens. The activation of PAR by trypsin or PAR-2-specific activating peptide (AP) caused an increase in cytosolic calcium, as well as the release of the CXC chemokines IL-8 and growth-related oncogene (GRO)-alpha, but not the release of CC chemokines or IL-6. AP-induced CXC chemokine was sensitive to PTX and activation of NF-kappaB was inhibited by BAY11-7085. Furthermore, a serine protease inhibitor significantly inhibited chemokine synthesis stimulated by trypsin and culture supernatants of staphylococci, whereas steroids and cysteine protease inhibitors had little effect.. PAR-2 plays a role in serine protease-mediated regulation - staphylococcal and non-staphylococcal origin - of IL-8 and GRO-alpha in nasal epithelial cells, but not in the regulation of CC chemokines. PAR-2 may therefore be involved in the pathophysiology of CRS and NP at different sites of activation, namely (i) proteases, (ii) the PAR-2 receptor itself or (iii) the application of novel agents that block NF-kappaB/IkappaB-alpha signalling.

Topics: Acute Disease; Adult; Aged; Bacterial Proteins; Calcium; Case-Control Studies; Cells, Cultured; Chemokine CCL11; Chemokine CCL17; Chemokine CCL5; Chemokine CXCL1; Chemokines, CC; Chemokines, CXC; Chronic Disease; Culture Media, Conditioned; Female; Humans; Interleukin-6; Interleukin-8; Male; Middle Aged; Nasal Mucosa; Nasal Polyps; NF-kappa B; Nitriles; Peptides; Pertussis Toxin; Receptor, PAR-2; Rhinitis; RNA, Messenger; Serine Endopeptidases; Serine Proteinase Inhibitors; Signal Transduction; Sinusitis; Staphylococcus aureus; Sulfones; Trypsin

The aim of this study was to evaluate the effects of Staphylococcus aureus exotoxin B (SE-B) on proinflammatory cytokine/chemokine releases in primary nasal epithelial cell cultures (NECC) of subjects with and without chronic rhinosinusitis (CRS).. NECC (CRS: n = 14;. n = 11) were stimulated with SE-B. Protein concentrations of interleukin-(IL)-1beta, IL-6, and IL-8 levels were measured in NECC supernatants by ELISA before (T0) and after 24 hr stimulation with SE-B (T1).. T0: supernatants of the NECC of CRS patients contained significant lower levels of IL-8 (2.1 ng/ml) compared to CONTROLS (IL-8: 6.2 ng/ml; P < 0.01). T1: SE-B induced a significant increase of IL-6 in NECC (P < 0.001). IL-1beta was not detectable.. This is the first study evaluating the effects of exotoxins on NECC. SE-B showed proinflammatory effects on NECC.. Our data suggest that resident NECC are involved in immunological responses to Staphylococcus aureus toxins, supplementing the so-called "superantigen hypothesis" in CRS.

Topics: Cells, Cultured; Chronic Disease; Culture Media, Serum-Free; Epithelial Cells; Exotoxins; Humans; Interleukin-1; Interleukin-6; Interleukin-8; Nose; Rhinitis; Sinusitis; Staphylococcus aureus; Superantigens

While various microorganisms have been recovered from patients with chronic rhinosinusitis, the inflammatory impact of virulence factors, in particular proteases from Staphylococcus aureus and coagulase negative staphylococci on the nasal epithelium, has not yet been investigated. Expression of CXC chemokines was determined in the epithelium of patients with chronic rhinosinusitis by immunohistochemistry. In a cell culture system of A549 respiratory epithelial cells, chemokine levels were quantified by enzyme-linked immunosorbent assay (ELISA) after stimulation with supernatants originating from three different staphylococcal strains or with trypsin, representing a serine protease. Inhibition experiments were performed with prednisolone, with the serine protease inhibitor 4-(2-aminoethyl)-benzenesulphonylfluoride (AEBSF) and with the nuclear transcription factor (NF)-kappaBeta inhibitor (2E)-3-[[4-(1,1-dimethylethyl)phenyl]sulphonyl]-2-propenenitrite (BAY) 11-7085. Electromobility shift assays (EMSA) were used to demonstrate NF-kappaB-dependent protein synthesis. CXC chemokines interleukin (IL)-8, growth-related oncogene alpha (GRO-alpha) and granulocyte chemotactic protein-2 (GCP-2) were expressed in the patients' epithelium whereas epithelial cell-derived neutrophil attractant 78 (ENA-78) was rarely detected. In A549 cells, chemokines IL-8, ENA-78 and GRO-alpha but not GCP-2 were induced by trypsin and almost equal levels were induced by staphylococcal supernatants. IL-8, GRO-alpha and ENA-78 synthesis was suppressed almost completely by AEBSF and BAY 11-7085, whereas prednisolone reduced chemokine levels differentially dependent on the supernatant added. CXC chemokines were detectable in the epithelium of patients with chronic rhinosinusitis. Staphylococcal serine proteases induced CXC chemokines in A549 cells, probably by the activation of proteases activated receptors, and thus might potentially be involved in neutrophilic inflammation in chronic sinusitis.

Topics: Adult; Aged; Cell Line; Chemokine CXCL1; Chemokine CXCL5; Chemokines, CXC; Chronic Disease; Epithelial Cells; Female; Humans; Immunity, Mucosal; Immunoenzyme Techniques; Intercellular Signaling Peptides and Proteins; Interleukin-8; Male; Metalloendopeptidases; Middle Aged; Nasal Mucosa; Neutrophil Infiltration; NF-kappa B; Nitriles; Prednisolone; Rhinitis; Serine Proteinase Inhibitors; Signal Transduction; Sinusitis; Sulfones; Trypsin

The aetiology of chronic rhinosinusitis (CRS) remains unclear. The purpose of this study was to investigate neutrophil-attracting chemokine patterns in CRS without nasal polyposis.. The biological activity of the chemokines was identified using a two-step high-performance liquid chromatography (HPLC) technique combined with a bioassay in extracts from 55 CRS patients, and in the turbinate mucosa (TM) of patients (N=51) undergoing septumplasty. The biologic activity of each chemokine was assessed using blocking antibodies to chemokines. Immunolocalization of detected neutrophil chemokines was performed by quantitative evaluation of immunohistochemistry. Besides, PCR analysis was performed to quantify neutrophil chemokine mRNA.. In CRS, the chemokines primarily detected by two-step HPLC were growth-related oncogene-alpha (GRO-alpha) and the granulocyte chemotactic protein-2 (GCP-2). Blocking of GCP-2 and GRO-alphad each resulted in chemotaxis inhibition rates of 43.3% and 35.9%, respectively, whereas anti-IL-8 and anti-ENA-78 had no effect. Both GCP-2 and GRO-alphad were generally synthesized by the surface epithelium and mucosal glands while GRO-alpha in particular was synthesized by endothelial cells, as shown by immunohistochemistry. The concentrations of the chemokines IL-8 and epithelial cell-derived neutrophil attractant-78 (ENA-78) were low in CRS and TM.. It appears that both GRO-alpha and GCP-2 contribute to neutrophil chemotaxis in CRS, whereas IL-8 and ENA-78 appear to be of secondary importance for the chemotaxis of neutrophils in this condition. The expression of chemokines in mucosal gland cells is the main phenomenon involved in constitutive neutrophil chemotaxis in the TM.

Topics: Adult; Chemokine CXCL1; Chemokine CXCL5; Chemokine CXCL6; Chemokines, CXC; Chemotaxis, Leukocyte; Chromatography, High Pressure Liquid; Enzyme-Linked Immunosorbent Assay; Female; Glyceraldehyde-3-Phosphate Dehydrogenases; Humans; Intercellular Signaling Peptides and Proteins; Interleukin-8; Male; Nasal Mucosa; Neutrophil Activation; Neutrophils; Reverse Transcriptase Polymerase Chain Reaction; Rhinitis; Sinusitis

To determine the role of interleukin (IL)-4, IL-4 receptor (R), IL-6, IL-8, IL-11, and transforming growth factor (TGF)-beta in chronic rhinosinusitis (CRS) and chronic rhinosinusitis with nasal polyposis (CRS/NP).. Sinus tissue from patients undergoing endoscopic sinus surgery for CRS and CRS/NP was collected. Sinus tissue was then analyzed using reverse-transcription polymerase chain reaction (RT-PCR) to detect transcription of IL-4R, IL-6, IL-8, and IL-11. Sinus tissue samples were also cultured in vitro, treated with IL-4 for 24 hours, and real-time PCR was used to quantify the transcription of TGF-beta.. Twenty patients were evaluated, 9 with CRS/NP and 11 with CRS alone. The mean age was 43 (20-74) years, with 13 females and 7 males. IL-4R, IL-6, IL-8, and IL-11 were identified by RT-PCR in all 20 patients. The transcription of TGF-beta was found to be 3.2 times greater in patients with CRS/NP than in patients with CRS alone (P = .047).. IL-6, IL-8, and IL-11 are nonspecific markers of sinus inflammation being transcribed in patients with CRS and patients with CRS/NP. However, patients with CRS/NP demonstrate increased transcription of TGF-beta in response to IL-4 treatment, suggesting an IL-4 mediated mechanism for stromal proliferation in the formation of nasal polyposis.

Topics: Adult; Aged; Biomarkers; Cell Proliferation; Chronic Disease; Endoscopy; Female; Humans; Interleukin-11; Interleukin-4; Interleukin-6; Interleukin-8; Interleukins; Male; Middle Aged; Nasal Polyps; Receptors, Interleukin-4; Reverse Transcriptase Polymerase Chain Reaction; Rhinitis; Sinusitis; Transforming Growth Factor beta

The characteristic feature of chronic rhinosinusitis with nasal polyposis (CRSwNP) is eosinophilic inflammation of the sinus mucosa; a type of inflammation also seen in asthmatic airways. Similar histopathologic findings of airway remodelling are present in both diseases. Remodelling is tightly controlled by matrix metalloproteinases (MMP). Increase of collagenase-2 (MMP-8) expression in the bronchial epithelial cells has been described in asthmatic patients, but it has not been studied in CRSwNP.. The concentrations and degree of activation of MMP-8 were analysed by immunofluorometric assay and Western blotting, respectively, in sinus mucus samples from CRSwNP patients and in nasal lavages from healthy controls in relation to inductive cytokines interleukin-8 (IL-8) and tumour necrosis factor-alpha (TNF-alpha).. Significantly elevated levels of MMP-8 and IL-8 but not TNF-alpha were found in CRSwNP patients relative to controls. In particular, the activation of mesenchymal-type MMP-8 but not polymorphonuclear-type MMP-8 was associated with elevated IL-8 levels.. The IL-8 and MMP-8 seemingly form an inductive cytokine-proteinase cascade in CRSwNP pathogenesis. Together they provide a target for novel therapies and a diagnostic tool for monitoring CRSwNP treatment.

Topics: Aged; Chronic Disease; Female; Humans; Interleukin-8; Male; Matrix Metalloproteinase 8; Middle Aged; Nasal Mucosa; Nasal Polyps; Paranasal Sinuses; Rhinitis; Sinusitis; Tumor Necrosis Factor-alpha; Up-Regulation

To investigate the correlation between nuclear factor-kappa B (NF-kappaB) activity and cytokine expression in nasal mucosa of chronic sinusitis.. IL-5, IL-6 and IL-8 levels in nasal mucosa were assayed by the method of ELISA in 52 cases of chronic sinusitis [concomitant with allergic rhinitis (AR group), without allergic rhinitis (NAR group)] and 12 normal subjects. Semi-quantitative RT-PCR and immunohistochemical staining were used to examine P50 and P65 subunits of NF-KB expressions and activation in nasal mucosa. The correlation between activities of NF-KB P50 and P65 subunits and cytokine expression was evaluated.. IL-5, IL-6 and IL-8 levels in both AR and NAR groups were significantly increased (all P < 0.01 for AR group; P < 0.05, 0.05, 0.01, respectively, for NAR group, as compared with normal group), and the levels were much higher in AR group than that in NAR group (P < 0.01, 0.05, 0.01, respectively). The levels of P50 and P65 mRNA in both AR and NAR groups were enhanced (all P < 0.01 for AR group; all P < 0.01 for NAR group, as compared with normal group), and AR group had markedly greater P50 and P65 mRNA levels in comparison with NAR group (both P < 0.05). Immunohistochemical study revealed that nucleus-present rates of P50 and P65 in both AR and NAR groups were significantly higher than those of control group (all P < 0.01), and they were much greater in AR group as compared with NAR group (all P < 0.01). Pearson correlation analysis demonstrated that P50 and P65 nucleus-present rates were closely correlated with IL-6 and IL-8 levels, but not IL-5. The correlation coefficient was 0. 49 for P50 and IL-6, 0. 54 for P50 and IL-8, 0. 61 for P65 and IL-6, and 0.66 for P65 and IL-8 (all P < 0.01).. Activation of P50 and P65 subunits of NF-kappaB might be one of the mechanisms for induction of IL-6 and IL-8 expression in chronic sinusitis. Concomitance of allergic rhinitis with chronic sinusitis further increased activities of NF-kappaB subunits, and further elevated IL-6 and IL-8 expression. IL-5 expression was independent of NF-kappaB pathway in chronic sinusitis.

Topics: Adult; Chronic Disease; Female; Humans; Interleukin-5; Interleukin-6; Interleukin-8; Male; Middle Aged; Nasal Mucosa; NF-kappa B p50 Subunit; Rhinitis; RNA, Messenger; Sinusitis; Transcription Factor RelA

The etiology and classification of chronic rhinosinusitis with and without nasal polyps still remain unclear. Based on investigations of inflammation type in biopsies from patients with nasal polyposis and chronic non-polypous rhinosinusitis, we tried to determine whether there is a need for further classification of chronic rhinosinusitis into two disease entities.. Biopsies of diffuse nasal polyposis (n= 37) and chronic rhinosinusitis without nasal polyps (n= 41) were examined for eosinophil and neutrophil tissue infiltration, degree of inflammation, and involved cytokines in inflammation mechanisms.. Neutrophil elastase positive neutrophils and CD38-positive lymphocytes were characterized in nasal polyposis and chronic non-polypous rhinosinusitis by immunohistochemistry. Using a monoclonal antibody against eosinophilic cationic protein (ECP) activated eosinophils were identified. In tissue homogenates, albumin was quantified as a marker for inflammation vascular permeability. In addition, interleukin (IL)-8 and IL-5 were determined by means of quantitative ELISA measurements in homogenates.. Significantly increased numbers of eosinophils and neutrophils were detected in nasal polyposis. In chronic rhinosinusitis without nasal polyps, tissue infiltration was dominated by lymphocytes and neutrophils. The concentration of albumin and IL-5 was significantly higher in nasal polyps than in chronic rhinosinusitis without nasal polyps. IL-8 protein levels did not differ significantly between the two tissue types. In addition, patients' durations of illness did not differ significantly.. Different types and quantities of inflammatory cells as well as different levels of inflammation support our hypothesis that there is need for further subdivision of chronic rhinosinusitis into two disease entities.

Topics: ADP-ribosyl Cyclase; ADP-ribosyl Cyclase 1; Adult; Albumins; Antigens, CD; Blood Proteins; Chronic Disease; Eosinophil Granule Proteins; Eosinophils; Female; Humans; Inflammation Mediators; Interleukin-5; Interleukin-8; Leukocyte Elastase; Lymphocytes; Male; Membrane Glycoproteins; Middle Aged; Nasal Polyps; Neutrophils; Rhinitis; Ribonucleases; Sinusitis; Tomography, X-Ray Computed

To investigate the effect of the proinflammatory cytokines, including IL-1, IL-8 and TNF in the pathogenesis of chronic sinusitis.. Using immunohistochemical streptavidin-biotin peroxide complex (SABC) method to investigate the expression of interleukin-1 (IL-1), IL-8 and tumor necrosis factor (TNF) in the local mucosa of the two type chronic sinusitis.. There were significantly large numbers of IL-1+, IL-8+ and TNF+ cell in chronic sinusitis type I compared with healthy control (P < 0.01). The numbers of IL-8+ and TNF+ cell in the chronic sinusitis type II were significantly larger than that in the healthy control (P < 0.01). The numbers of IL-1+ cell in chronic sinusitis type I were significantly larger than that in chronic sinusitis type II (P < 0.01).. This study indicates that proinflammatory cytokines may play an important role in the pathogenesis of chronic sinusitis. It appears that specific cytokine patterns are found in different forms of sinusitis. The investigation of different cytokine patterns may help to understand the different pathogenesis in chronic sinusitis subgroups.

Topics: Adolescent; Adult; Chronic Disease; Female; Humans; Interleukin-1; Interleukin-8; Male; Middle Aged; Mucous Membrane; Sinusitis; Tumor Necrosis Factor-alpha

To elucidate the mechanisms involved in the action of new macrolides on chronic sinusitis, we examined the effects of clarithromycin (CAM) and roxiythromycin (RXM) on the expression of adhesion molecules (L-selectin and Mac-1) on peripheral blood neutrophils of individuals with chronic sinusitis. The administration of CAM and RXM slightly inhibited the down-regulation of L-selectin expression on neutrophils induced by interleukin (IL)-8 stimulation. Furthermore, the administration of CAM strongly inhibited the IL-8-induced up-regulation of Mac-1 expression on neutrophils. These observations suggest that the new macrolides such as CAM and RXM may affect the functions of neutrophils in chronic sinusitis by modulating the expression of L-selectin and Mac-1 molecules on neutrophils, thereby attenuating the adhesion of neutrophils.

Topics: Adolescent; Adult; Aged; Chronic Disease; Clarithromycin; Down-Regulation; Female; Humans; Interleukin-8; L-Selectin; Macrophage-1 Antigen; Male; Middle Aged; Neutrophils; Roxithromycin; Sinusitis; Up-Regulation

Long-term, low-dose macrolide therapy is effective in the treatment of chronic rhinosinusitis. The mechanism of its anti-inflammatory effect and how this differs from corticosteroids remains unclear. The effect of clarithromycin and prednisolone on interleukin-5, interleukin-8, and granulocyte-macrophage colony-stimulating factor production by cultured chronic sinusitis nasal mucosa was examined in the study.. Nasal mucosa was obtained from 11 patients with chronic sinusitis. This tissue was cultured for 24 hours in the presence of clarithromycin or prednisolone at a variety of concentrations. Cytokine levels were determined by enzyme-linked immunoassay.. Clarithromycin and prednisolone each produced significant reductions in interleukin-5, interleukin-8, and granulocyte-macrophage colony-stimulating factor production. There was no significant difference between the effects of clarithromycin and prednisolone.. Macrolide antibiotics are capable of inhibiting pro-inflammatory cytokine production in vitro and are as potent as prednisolone. This mechanism is likely to be at least partly responsible for the clinical efficacy of macrolide antibiotics in chronic rhinosinusitis.

Topics: Adult; Anti-Bacterial Agents; Anti-Inflammatory Agents; Chronic Disease; Clarithromycin; Culture Techniques; Cytokines; Enzyme-Linked Immunosorbent Assay; Female; Glucocorticoids; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Interleukin-5; Interleukin-8; Male; Middle Aged; Nasal Mucosa; Prednisolone; Rhinitis; Sinusitis

In rhinologic disorders such as polyposis or rhinitis, nasal cytology allows differentiation between patients according to the degree of eosinophilia in nasal secretions. The egress of eosinophil and/or neutrophil polymorphonuclears from the underlying mucosa might correlate with the release of soluble mediators of cell activation such as the chemokine IL-8, and such molecules of the innate immunity as the LPS-receptor CD14 or lysozyme. We assayed the levels of these three molecules in nasal secretions in correlation with cytologic findings and especially the degree of eosinophilia.. Fifty-four patients from a prospective study of nasal secretions were enrolled in this work. They constituted two groups of 27 patients each, respectively, with or without more than 20% eosinophils in nasal secretions. Nasal secretions were collected by aspiration, weighed and diluted in a fixed amount of buffer. Classic cytologic analyses were performed on the pelleted cells and IL-8, sCD14, and lysozyme levels were assayed in the cell-free supernatants.. Cytologic analyses included cell-enumeration in Neubauer's chambers, and differentials performed on May-Grünwald Giemsa-stained cytospins. ELISA tests were used to assay the levels of IL-8 and sCD14. Lysozyme concentrations were assayed in immuno-nephelometry.. Significantly lower levels of IL-8 and sCD14 were observed in patients with eosinophilia than in patients with a predominance of neutrophils, whereas no difference was observed in lysozyme concentrations.. These data show that the egress of neutrophils in nasal secretions is associated with high levels of IL-8 and sCD14.

Topics: Adolescent; Adult; Aged; Eosinophilia; Female; Humans; Inflammation Mediators; Interleukin-8; Leukocyte Count; Lipopolysaccharide Receptors; Male; Middle Aged; Muramidase; Nasal Lavage Fluid; Nasal Mucosa; Nasal Polyps; Neutrophils; Reference Values; Rhinitis; Sinusitis

Low-dose, long-term macrolide treatment has recently been reported to be very effective in patients with chronic airway diseases. We examined the in vivo and in vitro effects of 14-membered macrolide antibiotics erythromycin (EM) and clarithromycin (CAM) on interleukin (IL)-8 secretion from human nasal epithelial cells. Fifteen patients with chronic sinusitis received macrolide treatment (CAM 400 mg/day) for 1 to 3 months. The number of infiltrated neutrophils and IL-8 concentrations in the nasal discharges of these patients decreased significantly at 1 to 2 months after the treatment. In vitro effects of EM and CAM on IL-8 secretion were examined in nasal epithelial cells cultured at the air-liquid interface. After 14-day culture in the air-liquid interface, macrolide antibiotics were added in medium for 24 h. EM and CAM at concentrations of 10(-4) M did not affect spontaneous secretions or IL-1 beta-induced secretions of IL-8 either apically or basolaterally. When cells were preincubated with 10(-4) M CAM for 7 days, the IL-1 beta-induced secretion of IL-8 decreased significantly. However, no difference was observed between the effects of 10(-4) M CAM and 10(-4) M josamycin, a 16-membered macrolide. These results suggest that macrolide treatment inhibits neutrophil infiltration and IL-8 secretion in nasal epithelium in vivo and that these clinical effects depend on a mechanism other than the direct action of macrolide on nasal epithelial cells.

Topics: Adolescent; Adult; Aged; Anti-Bacterial Agents; Cells, Cultured; Child; Chronic Disease; Clarithromycin; Drug Administration Schedule; Epithelium; Erythromycin; Humans; Interleukin-1; Interleukin-8; Middle Aged; Nasal Mucosa; Neutrophils; Nose; Sinusitis

Epithelial hyperplasia and mucosal infiltration of leukocytes are common features of chronic rhinosinusitis. The epithelium can produce chemoattractant cytokines that may contribute to leukocyte infiltration in rhinosinusitis.. We sought to determine whether mucosal IL-8 gene expression is increased in chronic rhinosinusitis and to relate IL-8 gene expression to disease severity.. We used both a noncompetitive and a quantitative, competitive reverse transcription-polymerase chain reaction to examine IL-8 gene expression in samples of sinus mucosal tissue obtained during surgery from 22 patients with chronic rhinosinusitis and 9 normal control subjects. IL-8 gene expression was related to disease severity assessed by sinus computed tomography (CT) scores and to symptom scores assessed by means of a questionnaire.. Sinus mucosal IL-8 gene expression was not detected in any of the control subjects but was present in 12 of 22 (55%) patients with rhinosinusitis. Sinus CT scores and symptom scores were both significantly higher in patients with positive mucosal IL-8 gene expression than in subjects with no detectable IL-8 gene expression. Positive IL-8 gene expression was not predicted by history of prior surgery nor by atopic or asthmatic status. In 9 subjects with positive IL-8 gene expression, levels of mRNA expression, assessed by competitive reverse transcription-polymerase chain reaction, correlated significantly (rho = 0.72, P <.05) with sinus CT scores.. Sinus mucosal expression of the gene for IL-8 is increased in patients with chronic rhinosinusitis, and the level of IL-8 gene expression correlates with disease severity.

Topics: Adult; Aged; Asthma; Chronic Disease; Epithelial Cells; Female; Gene Expression; Humans; Hypersensitivity, Immediate; Interleukin-8; Male; Middle Aged; Mucous Membrane; Reverse Transcriptase Polymerase Chain Reaction; Rhinitis; RNA, Messenger; Sinusitis; Tomography, X-Ray Computed

The mechanism of the efficacy of long-term low-dose macrolide therapy for chronic sinusitis is not fully understood. The authors studied the inhibitory effect of erythromycin on interleukin-8 (IL-8) secretion from exudative cells in the nasal discharge of patients with chronic sinusitis.. Exudative cells in the nasal discharge were isolated from six patients with nonallergic chronic sinusitis. The cells, more than 90% of which were neutrophils, were incubated with or without erythromycin in the presence of 10 micrograms/mL of lipopolysaccharide. The IL-8 concentrations in the culture supernatants were measured by enzyme-linked immunoassay.. The amount of secreted IL-8 in the absence of erythromycin was 682 +/- 226 pg/10(6) cells/24 h. The IL-8 secretion was significantly reduced to 66 +/- 15% and 46 +/- 13% of the control in the presence of 10(-6) and 10(-5) M of erythromycin, respectively.. Erythromycin may act as a biologic modulator that inhibits IL-8 secretion from exudative cells and thereby blocks the vicious circle of neutrophil recruitment and IL-8 generation in the inflammatory site in chronic sinusitis.

Topics: Anti-Bacterial Agents; Chronic Disease; Dose-Response Relationship, Drug; Enzyme-Linked Immunosorbent Assay; Erythromycin; Humans; In Vitro Techniques; Interleukin-8; Long-Term Care; Nasal Mucosa; Neutrophil Activation; Neutrophils; Sinusitis

Topics: Anti-Bacterial Agents; Chronic Disease; Clarithromycin; Female; Humans; Interleukin-8; Leukocyte Count; Male; Middle Aged; Nasal Mucosa; Neutrophils; Sinusitis

We have recently phenotyped inflammation in non-infectious allergic and non-allergic chronic maxillary sinusitis using sinus biopsies and lavage fluids. In this first paper, we have concentrated our work on the eosinophil, T cell, mast cell and macrophage infiltrates. However, many unresolved questions remain and particularly the role of neutrophils needed to be addressed. In the present study, we focused on the neutrophilic inflammation: myeloperoxidase (MPO) and interleukin-8 (IL-8) were measured by immunoassays and neutrophils were enumerated by conventional staining in the sinus lavage fluids of 16 patients with chronic sinusitis and six control subjects. Both MPO and IL-8 levels were significantly higher in patients than in controls (P < 0.01 and 0.005, respectively). There was a significant correlation between MPO levels and neutrophil numbers, and between MPO and IL-8 levels in the sinus lavage fluid (P < 0.0001, Spearman rank correlation). The presence of high levels of IL-8 in the lavage fluids of patients suffering from chronic sinusitis, levels which correlate with those of MPO, suggests that this cytokine may activate neutrophils in this chronic disease.

Topics: Adolescent; Adult; Aged; Chronic Disease; Female; Humans; Interleukin-8; Male; Middle Aged; Nasal Lavage Fluid; Peroxidase; Phenotype; Sinusitis

Effects of long-term low-dose macrolide administration were studied in patients with chronic sinusitis. Twelve patients with non-allergic chronic sinusitis were orally given 150 mg roxithromycin once a day without other treatments. The patients underwent computed tomography before and after the treatment, and paranasal sinus aeration was analyzed quantitatively. The number of neutrophils in the nasal smear was semiquantitatively assessed on a grading scale, and the IL-8 concentration in the nasal discharge was measured by enzyme immunoassay. The aeration of all four sinuses significantly improved, and recruited neutrophils and the IL-8 level in the nasal discharge were simultaneously reduced after the treatment. These findings suggest that long-term low-dose roxithromycin administration inhibits the positive feedback mechanism of neutrophil recruitment and IL-8 production by the recruited neutrophils, which is considered to be an essential cause of the prolongation of sinusitis.

Topics: Administration, Oral; Adolescent; Adult; Aged; Anti-Bacterial Agents; Chronic Disease; Female; Humans; Immunity, Mucosal; Interleukin-8; Male; Middle Aged; Nasal Mucosa; Neutrophil Activation; Neutrophils; Roxithromycin; Sinusitis; Time Factors

The mechanism of macrolide therapy in chronic sinusitis patients is unclear. The authors studied the effect of macrolides on interleukin (IL)-8 secretion from cultured human nasal epithelial cells. Epithelial cells harvested from the nasal polyps of patients with chronic sinusitis were primary-cultured, and secreted IL-8 in culture media was measured by enzyme immunoassay. The cells secreted considerable amounts of IL-8 constitutively and in response to lipopolysaccharide. The secretion was significantly inhibited by 10(-5) M of erythromycin, clarithromycin, roxithromycin, and josamycin. 10(-6) M erythromycin still showed the inhibitory effect, whereas the same concentration of josamycin did not. These results indicate that macrolide antibiotics may act as an immunomodulator to reduce IL-8 in inflammatory sites and, at least partially, account for the clinically discrepant effects between 14- and 16-membered ring macrolides in long-term low-dose therapy for chronic sinusitis.

Topics: Adult; Aged; Anti-Bacterial Agents; Cell Culture Techniques; Chronic Disease; Dose-Response Relationship, Drug; Endoscopy; Epithelial Cells; Female; Humans; Immunoenzyme Techniques; Interleukin-8; Macrolides; Male; Middle Aged; Nasal Mucosa; Nasal Polyps; Sinusitis

Topics: Animals; Anti-Bacterial Agents; Chemotaxis, Leukocyte; Chronic Disease; Female; Humans; Interleukin-8; Male; Middle Aged; Nasal Mucosa; Neutrophils; Rats; Rats, Sprague-Dawley; Roxithromycin; Sinusitis

The mechanism of neutrophil recruitment in patients with chronic sinusitis is unclear.. This study aims to elucidate the role of IL-8 in inducing neutrophil accumulation in the nasal discharge of patients with chronic sinusitis.. Nasal discharge and mucosal specimens were obtained from two groups of patients, those with chronic sinusitis and those with allergic rhinitis. The samples were subjected to immunohistochemical examination and in situ hybridization. The IL-8 level in the nasal discharge was measured by enzyme immunoassay.. Immunoreactivity to IL-8 was observed in polymorphonuclear cells of nasal smear, in nasal gland duct cells, and in epithelial cells of the chronic sinusitis group; whereas those of the allergic rhinitis group mostly showed little or no reaction. Similar patterns of localization were shown by in situ hybridization for IL-8 messenger RNA. The IL-8 level in nasal discharge was significantly higher in the chronic sinusitis group than in the allergic rhinitis group.. These results suggest that chemotactic factors in sinus effusion, including IL-8 derived from nasal gland duct cells and epithelial cells, attract neutrophils out of mucosa, and the neutrophils that have emigrated into the sinus effusion secrete IL-8. This induces further neutrophil accumulation in the sinus effusion of patients with chronic sinusitis.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Cell Movement; Child; Child, Preschool; Chronic Disease; Female; Humans; Immunohistochemistry; Interleukin-8; Male; Middle Aged; Nasal Mucosa; Neutrophils; RNA, Messenger; Sinusitis

Fronto-ethmoidal mucocoeles have the capacity to destroy bone. Sinus lining tissue has been obtained at surgery from patients with mucocoeles, from those with chronic sinusitis undergoing endoscopic sinus surgery and from patients undergoing craniofacial resection. Tissues have been frozen, sectioned, and subjected to immunohistochemical examination with monospecific antibodies for the presence of the potent osteolytic cytokines interleukins-1 and -6 and tumour necrosis factor alpha. In addition, the chemotactic intercrine--interleukin-8 was investigated. The presence of the cytokine-inducible vascular endothelial adhesion receptors--Inter-Cellular Adhesion Molecule (ICAM)-1 and E-Selectin (also known as Endothelial Leukocyte Adhesion Molecule--ELAM) was also determined. Normal sinus tissue showed no immunoreactivity with the antibodies to these various moieties. Surprisingly, only a small proportion of tissues from patients with chronic sinusitis showed the presence of cytokines or vascular adhesion receptors. In contrast, all specimens of fronto-ethmoidal mucocoeles showed positive staining for IL-1 alpha and beta and for ICAM-1 and E-selectin. IL-1 immunostaining was restricted to the epithelial cell population not being found in infiltrating leukocytes. In 40% of mucocoeles infiltrating macrophage-like cells showed the presence of tumour necrosis factor alpha. The presence of the potent osteolytic cytokine--IL-1 in all specimens of fronto-ethmoidal mucocoeles coupled to the finding of the IL-1-inducible adhesion molecules ICAM-1 and E-Selectin argues strongly that IL-1 is released from the epithelial cells and that this cytokine may be the factor causing the erosion of bone overlying the expanding mucocoele. The nature of the signals inducing cytokine synthesis remain, however, unidentified.

Topics: Adult; Aged; Antigens, Differentiation, Myelomonocytic; Cell Adhesion Molecules; Chemotactic Factors; Cytokines; E-Selectin; Ethmoid Sinus; Female; Frontal Sinus; Humans; Intercellular Adhesion Molecule-1; Interleukin-1; Interleukin-6; Interleukin-8; Macrophages; Male; Membrane Glycoproteins; Middle Aged; Mucocele; Mucous Membrane; Paranasal Sinus Diseases; Platelet Endothelial Cell Adhesion Molecule-1; Receptors, Immunologic; Sinusitis; Tumor Necrosis Factor-alpha