interleukin-8 and Nasal-Polyps

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

Recent studies indicated that interleukin (IL)-17, growth-related oncogene (GRO)-α and IL-8 play an important role in the pathogenesis of nasal polyps. However, the effects of the increased amount of IL-17 and the production of GRO-α and IL-8 in human nasal polyp fibroblasts are not completely understood. This study aimed to determine the effects of the increased IL-17 on the changes of GRO-α and IL-8 expression in human nasal polyp fibroblasts and further investigate the mechanism of neutrophil infiltration in nasal polyps. Nasal polyp fibroblasts were isolated from six cases of human nasal polyps, and the cells were stimulated with five different concentrations of IL-17. Real-time fluorescence quantitative polymerase chain reaction (RT-PCR) was used to detect the mRNA expression of GRO-α and IL-8. The mRNA of GRO-α and IL-8 was expressed in unstimulated controls and remarkably increased by stimulation with IL-17. Moreover, the levels of GRO-α and IL-8 produced by fibroblasts were increased gradually with the increases in IL-17 concentrations. The present study showed that nasal fibroblasts can produce GRO-α and IL-8, and their production is remarkably enhanced by IL-17 stimulation, thereby clarifying the mechanism of the IL-17 mediated neutrophil infiltration in nasal polyps. These findings might provide a rationale for using IL-17 inhibitors as a treatment for nasal inflammatory diseases such as nasal polyps.

Topics: Adult; Cells, Cultured; Chemokine CXCL1; Female; Fibroblasts; Humans; Interleukin-17; Interleukin-8; Male; Middle Aged; Nasal Polyps; Neutrophil Infiltration; RNA, Messenger

Although chronic rhinosinusitis (CRS) is one of the most frequently reported chronic diseases its etiology is not well understood. Recently, fungi have been proposed to influence the chronicity of rhinosinusitis. If fungi do play an important role then topical antifungal treatment may improve the inflammatory process of CRS. Therefore, in this study we measured inflammatory cytokine levels in nasal polyps after intranasal antifungal irrigation.. Nasal polyps were collected before and 4 weeks after treatment with 100 mg/l topical amphotericin B (n = 16), 50 mg/l topical amphotericin B (n = 14) or normal saline (n = 11). The cytokine--IL-5, IL-8, interferon-gamma, RANTES--protein content of polyp homogenates were determined by means of ELISA.. Nasal polyps were found to contain large amounts of cytokines (IL-5, IL-8 and RANTES) compared with normal inferior turbinates. After 4 weeks of treatment with topical agents, IL-5 levels tended to decrease in comparison with those of the other cytokines, but this difference was not statistically significant.. Topical amphotericin B treatment and nasal saline irrigation both influence the expression of nasal polyp cytokines. Topical nasal irrigation may influence the inflammatory process of CRS.

Topics: Administration, Intranasal; Administration, Topical; Adult; Amphotericin B; Antifungal Agents; Chemokine CCL5; Cytokines; Female; Humans; Interferon-gamma; Interleukin-5; Interleukin-8; Male; Mycoses; Nasal Polyps; Sodium Chloride; Therapeutic Irrigation

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

Chronic rhinosinusitis (CRS) is the most common immunological ENT disease, which has several phenotypes. The heterogeneity of CRS is due to the peculiarities of their pathogenetic mechanisms - the system of cytokines plays the crucial significance. They are biologically active substances and present regulatory peptides that demonstrate an immunomodulatory and regulatory effects not only in the local level but the system level as well.. To determine specific features of the cytokine profile in blood serum among patients with CRS without polyps (CRSwP).. Serum cytokines (IL-1α, IL-1RA, IL-1β, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-12p40, IL-12p70, IL-13, IL-15, IL-17, IFN-α2, IFN-γ, GM-CSF) were defined in 75 patients: 32 patients with CRSwP were operated (main group) - 17 with cyst of maxillary sinus, 15 with edema of the maxillary sinuses. The control group - 43 patients were under surgery for a deviated nasal septum (septoplasty). The groups were comparable to gender and age.. The cytokines detection rate was different in all groups. IL-4 (detection rate 93.3-95.3%) and IFN-γ (79.1-86.7%) were measured nearly the all groups. IL-8 (73.3-76.5%) and IL-17 (76.5-80.0%) were often measured in the group with CRSwP; in contrast to the control group - these indicators were lower: 60.5% and 65.5%, respectively. IL-1α (82.4%) and IFN-α2 (76.5%) were often detected in CRS with cystic formations. IL-1β, IL-5, IL-7, IL-9, IL-15 were measured in all groups in less than 30% of patients; IL-2 and IL-6 - in the group of CRS with cystic formation; IL-10, IL-12p40, IL-13 in the group with edema of maxillary sinuses. In a quantitative comparison of the concentration of cytokines. Significant differences in concentration of cytokines between the groups were not obtained (. CRSwP with cystic formation is characterized by the development of T2 type immune response and a higher inflammation-related tissue damage.. Хронический риносинусит (ХРС) — наиболее распространенное иммунологическое заболевание, имеющее несколько фенотипов. Гетерогенность ХРС обусловлена особенностями его патогенетических механизмов, ключевое значение в которых имеет система цитокинов — биологически активных веществ, представляющих собой регуляторные пептиды, оказывающие иммуномодулирующее и регуляторное действие на клетки организма как на местном, так и на системном уровнях.. Определить особенности цитокинового профиля в сыворотке крови у пациентов с ХРС без полипов (ХРСбП).. У 75 пациентов исследовано содержание цитокинов (IL-1α, IL-1RA, IL-1β, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-12р40, IL-12р70, IL-13, IL-15, IL-17, IFN-α2, IFN-γ, GM-CSF) в сыворотке крови. Из числа обследованных 32 пациентам (основная группа) проведено оперативное лечение по поводу ХРСбП: из них 17 пациентов были с кистоподобным образованием в верхнечелюстной пазухе, 15 пациентов — с отеком слизистой оболочки верхнечелюстных пазух; 43 пациента (группа сравнения) прооперированы по поводу искривленной перегородки носа (септопластика). Группы сопоставимы по полу и возрасту.. Частота обнаружения цитокинов в сыворотке крови пациентов в исследуемых группах была различной. Практически у всех пациентов обеих групп определены IL-4 (частота обнаружения 93,3—95,3%) и IFN-γ (79,1—86,7%). У пациентов с ХРСбП часто выявлялись IL-8 (73,3—76,5%) и IL-17 (76,5—80,0%), у пациентов группы сравнения эти показатели были ниже: 60,5% и 65,5% соответственно. При ХРС с кистоподобным образованием часто определялись IL-1α (82,4%) и IFN-α2 (76,5%). Сравнительно редко, менее чем у 30% пациентов всех групп, определялись IL-1β, IL-5, IL-7, IL-9, IL-15; у пациентов с ХРС с кистоподобным образованием — IL-2 и IL-6, у пациентов с пристеночным отеком слизистой оболочки — IL-10, IL-12р40, IL-13. При количественном сравнении концентраций интерлейкинов статистически значимые различия между группами не получены (. Хронический риносинусит без полипов с формированием кистоподобного образования характеризуется развитием иммунного ответа по типу Т2 с большей интенсивностью воспалительного процесса и более выраженным повреждением тканей.

Topics: Cytokines; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Interleukin 1 Receptor Antagonist Protein; Interleukin-10; Interleukin-12 Subunit p40; Interleukin-13; Interleukin-15; Interleukin-17; Interleukin-2; Interleukin-3; Interleukin-4; Interleukin-5; Interleukin-6; Interleukin-7; Interleukin-8; Interleukin-9; Nasal Polyps

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

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

Topics: Adult; Alternaria; Apamin; Aspergillus; Bee Venoms; Cell Proliferation; Cells, Cultured; Collagen Type I; Extracellular Matrix; Female; Fibroblasts; Humans; Interleukin-6; Interleukin-8; Male; Matrix Metalloproteinase 9; Melitten; Middle Aged; Nasal Polyps; p38 Mitogen-Activated Protein Kinases; Smad2 Protein; Smad3 Protein; Tissue Inhibitor of Metalloproteinase-1

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

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 (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

To investigate the distribution of mast cells in nasal polyps.. Biopsy specimens from patients with nasal polyps (n = 20) and control patients (n = 8) were obtained and included in this study. The distribution of mast cells in nasal polyps and the expression of chemokines (CCL5, CCL11, CX3CL1, IL-8, IL-6) in the epithelial cells of normal nasal mucosa and nasal polyps was determined by immunohistochemistry.. Mast cells migrate to intraepithelial in nasal polyps and the expression of chemokines (CCL5, CCL11, CX3CL1, IL-8) was up regulated in the epithelial cells of nasal polyps compare to normal nasal mucosa.. Our findings showed that mast cells migrate to intraepithelial in nasal polyps and the over expression of chemotaxins (CCL5, CCL11, CX3CL1, IL-8) may be response for mast cells' migration in nasal polyps. Mast cells might be associated with the development of nasal polyps.

Topics: Chemokine CCL11; Chemokine CCL5; Chemokine CX3CL1; Epithelial Cells; Humans; Immunohistochemistry; Interleukin-6; Interleukin-8; Mast Cells; Nasal Mucosa; Nasal Polyps; Up-Regulation

Nasal polyposis, a chronic inflammatory disease affecting the upper airways, is a valuable and accessible model to investigate the mechanisms underlying chronic inflammation. The main objective of this study was to investigate a potential involvement of the unfolded protein response (UPR) in the context of oxidative stress and inflammation in nasal epithelial cells from nasal polyps (NP).. Epithelial cells from NP (n = 20) and normal mucosa (Controls, n = 15) in primary culture were analyzed by global proteomic approach and cell biology techniques for the glucose-regulated protein 78 (GRP78), the spliced X-box-binding protein 1 (sXBP-1), the glucose-regulated protein 94 (GRP94), and the calreticulin (immunoblot, mass spectrometry, immunocytochemistry).. Proteomics analysis of human nasal epithelial cells in culture revealed the activation of the unfolded protein response in NP. Systematic cell biology and biochemical analysis of two markers (GRP78, sXBP-1) in the presence and absence of oxidative stress in NP showed a susceptibility of the unfolded protein response to oxidative stress compared to controls at least partially linked to an abnormal redox state of the protein disulfide-isomerase 4. This unfolded protein response was correlated with mitochondrial depolarization and secretion of interleukin 8 (IL-8) and leukotriene B4 (LTB4) and was prevented by mitochondrial antioxidant.. We show the existence of UPR in nasal epithelial cells that is linked to oxidative stress leading to IL-8 and LTB4 secretions. These mechanisms may participate in chronic inflammation in nasal polyposis.

Topics: Antioxidants; Cells, Cultured; Endoplasmic Reticulum Chaperone BiP; Epithelial Cells; Gene Expression Regulation; Humans; Inflammation; Interleukin-8; Leukotriene B4; Nasal Mucosa; Nasal Polyps; Oxidative Stress; Proteome; Proteomics; Unfolded Protein Response

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

Recent studies have shown that mast cells are involved in pathophysiologic processes of chronic inflammation. However, little is known about the distribution of mast cells in nasal polyps, which is a chronic inflammatory disease of the upper airways. Biopsy specimens from patients with nasal polyps (n = 20) and control patients without nasal polyps (n = 8) were included in this study. The distribution of mast cells in nasal polyps was determined by immunohistochemistry. Meanwhile, we detected the expression of chemokines (CCL5, CCL11, CX3CL1, IL-8, IL-6) in the epithelial cells of normal nasal mucosa and nasal polyps. In addition, the expression of these chemokines was investigated by western bolting in airway epithelial cells line (A549 cells) under inflammatory condition. Mast cells migrated toward intraepithelium in nasal polyps and the expression of chemokines (CCL5, CCL11, CX3CL1, IL-8) was up-regulated in the epithelial cells of nasal polyps compared with normal nasal mucosa. The expression of chemokines was also up-regulated in A549 cells after Lipopolysaccharide (LPS)-treatment for 3 hr and 6 hr. Our findings showed that mast cells migrate toward intraepithelium in nasal polyps and the overexpression of chemokines (CCL5, CCL11, CX3CL1, IL-8) suggested that they might be responsible for mast cells migration. It implies that mast cell play potential roles in the development of nasal polyps.

Topics: Adolescent; Adult; Aged; Biomarkers; Biopsy; Case-Control Studies; Cell Line, Tumor; Chemokine CCL11; Chemokine CCL5; Chemokine CX3CL1; Chemotaxis; China; Female; Humans; Immunohistochemistry; Interleukin-6; Interleukin-8; Lipopolysaccharides; Male; Mast Cells; Middle Aged; Nasal Mucosa; Nasal Polyps; Respiratory Mucosa; Time Factors; Up-Regulation; Young Adult

Treatment with glucocorticoids (GCs) is the cornerstone of nasal polyp (NP) therapy, but some patients respond poorly to them. Fibroblasts are involved in both inflammation and remodelling of NP. We aimed to evaluate whether NP fibroblasts are less sensitive to GCs' anti-proliferative and anti-inflammatory effects, compared to nasal mucosa (NM) fibroblasts.. Fibroblasts were obtained from NP (n = 8) from asthmatic patients undergoing endoscopic surgery and NM (n = 8) from patients undergoing nasal corrective surgery. Fibroblasts were stimulated with DMEM at 0.5% or 5% FBS, or TGF-β (5 ng/ml), with or without dexamethasone (10(-11) to 10(-5)M) for different times. Cell proliferation, collagen mRNA expression and IL-6 and IL-8 release were measured.. After 3-days, dexamethasone dose-dependently inhibited proliferation of NM (p < 0.001) but not that of NP fibroblasts. Dexamethasone (10(-6)M) reduced by 25% the proliferation of NM fibroblasts. Dexamethasone also inhibited proliferation of NM (p < 0.01) but not that of NP fibroblasts at 5-days. TGF-β induced collagen-1α1, -1α2, and -3α1 mRNA levels in both NM and NP fibroblasts (p < 0.05), and dexamethasone did not alter TGF-β-induced collagen mRNA levels in either fibroblast type at 24 h. Dexamethasone dose-dependently decreased (p < 0.05) FBS-induced IL-6 and IL-8 release in both NM and NP fibroblasts at 4 h, although at 10(-8)M, dexamethasone inhibited cytokine production in NM (p < 0.05) but not in NP fibroblasts.. This impaired sensitivity of nasal polyp fibroblasts to in vitro glucocorticoid effects concurs in part with the poor clinical response that these nasal polyp patients show to glucocorticoid treatment.

Topics: Adult; Cell Proliferation; Collagen; Dexamethasone; Female; Fibroblasts; Glucocorticoids; Humans; Interleukin-6; Interleukin-8; Male; Nasal Mucosa; Nasal Polyps; RNA, Messenger; Transforming Growth Factor beta

Although antihistamines and topical corticosteroids are used in combination to treat allergic rhinitis, their additive effect has not been yet demonstrated. The aim was investigate the antiinflammatory additive effect of mometasone and desloratadine on cytokine and sICAM-1 secretion by epithelial cells, and on eosinophil survival stimulated by human epithelial cells secretions from nasal mucosa and polyps.. Epithelial cells obtained from nasal mucosa or polyps were stimulated with 10% fetal bovine serum in presence of mometasone (10(-11) M-10(-5) M) with/without desloratadine (10(-5) M). Cytokine and sICAM-1 concentrations in supernatants were measured by ELISA. Peripheral blood eosinophils were incubated during 4 days with epithelial cell secretions with (10(-11) M-10(-5) M) and/or desloratadine (10(-5) M) and survival assessed by Trypan blue. Results are expressed as percentage (mean ± SEM) compared to control.. Fetal bovine serum stimulated IL-6, IL-8, GM-CSF and sICAM-1 secretion. In mucosa and polyp epithelial cells, mometasone inhibited this induced secretion while desloratadine inhibited IL-6 and IL-8. The combination of 10(-5) M desloratadine and 10(-9) M mometasone reduced IL-6 secretion (48 ± 11%, p < 0.05) greater extent than mometasone alone (68 ± 10%) compared to control (100%). Epithelial cell secretions induced eosinophil survival from day 1 to 4, this effect being inhibited by mometasone. At day 4, the combination of mometasone (10(-11) M) and desloratadine (10(-5) M) provoked an increased inhibition of eosinophil survival induced by cell secretions (27 ± 5%, p < 0.01) than mometasone (44 ± 7%) or desloratadine (46 ± 7%) alone.. These results suggest that the combination of desloratadine and mometasone furoate have a greater antinflammatory effect in an in vitro model of eosinophil inflammation than those drugs administered alone.

Topics: Adult; Aged; Aged, 80 and over; Anti-Inflammatory Agents; Cell Survival; Cells, Cultured; Cytokines; Dose-Response Relationship, Drug; Eosinophils; Epithelial Cells; Female; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Intercellular Adhesion Molecule-1; Interleukin-6; Interleukin-8; Loratadine; Male; Middle Aged; Mometasone Furoate; Nasal Mucosa; Nasal Polyps; Paracrine Communication; Pregnadienediols; Time Factors

Nasal polyposis (NP) is considered a subgroup within chronic rhinosinusitis. NP can be further subdivided into aspirin sensitive- and aspirin tolerant types (ASNP/ ATNP). Although the true etiology of NP has not been identified so far, it is agreed that NP represents an inflammatory disease of the nasal mucosa. Alterations of cellular kinase activities including that of IKK-2 might play a role in this inflammatory process.. Paraffin sections of ASNP, ATNP and controls were immunostained with Phospho-IkB-α antibody that detects the direct IKK-2 product (IkB-α. Intensity of epithelial staining was analysed semi-quantitatively by two independent observers. In cultured nasal polyp epithelial cells (NPECs) epithelial derived cytokines IL-8 and GRO α were induced by TNF-α or Staphylococcal supernatants and subsequently repressed by IKK-2 inhibitor TPCA-1.. Significant Phospho-IkB-α staining was observed in the nasal epithelium of ASNP compared to ATNP and controls suggesting strong IKK-2 activation in patients with ASNP in vivo. In vitro, pro-inflammatory cytokines IL-8 and GRO-α in NPECs were significantly repressed by TPCA-1.. IKK-2 activity is increased in the subgroup of ASNP. IL-8 and GRO-α responses were repressed by IKK-2 inhibitor TPCA-1 in vitro. IKK-2 inhibitors might represent a potential target for anti-inflammatory intervention in ASNP.

Topics: Adult; Aged; Amides; Chemokine CXCL1; Female; Humans; Immunohistochemistry; In Vitro Techniques; Interleukin-8; Male; Middle Aged; Nasal Mucosa; Nasal Polyps; NF-kappa B; Thiophenes

In vitro culture of nasal polyp cells is frequently used in the investigation of inflammatory mechanisms and effect of treatments in nasal polyposis. Research outcomes may, however, be influenced by the culture methodology used.. Nasal polyp and nasal mucosa in vitro fibroblast cultures were pre-treated with foetal bovine serum (FBS)-free culture medium or medium supplemented with either FBS or charcoal-stripped (cs) FBS. Cells were then stimulated with FBS or csFBS, with or without different doses of dexamethasone for 4 and 24h. IL-6, IL-8, GM-CSF and VEGF release and cell viability were measured.. The highest cytokine levels were found in growth-arrested cells stimulated with 10% FBS. csFBS poorly stimulated cytokine release. Nasal polyp released larger IL-8 amounts than nasal mucosa fibroblasts. Dexamethasone decreased cytokine production dose- and time-dependently in both nasal mucosa and nasal polyp fibroblasts. The IC25 of IL-8 inhibition by dexamethasone was higher in nasal polyp than in nasal mucosa fibroblasts. Cell viability did not differ among treatments.. Cytokine production by in vitro cultured nasal fibroblasts is affected by the culture conditions used and is inhibited by dexamethasone in both fibroblast types. Our results highlight the importance of culture methodology on nasal polyp research outcomes.

Topics: Anti-Inflammatory Agents; Cell Culture Techniques; Cell Survival; Cytokines; Dexamethasone; Dose-Response Relationship, Drug; Fibroblasts; Humans; Inflammation Mediators; Interleukin-6; Interleukin-8; Nasal Mucosa; Nasal Polyps; Vascular Endothelial Growth Factor A

Cystic fibrosis is an autosomal recessive disorder and it is characterised by chronic bacterial airway infection which leads to progressive lung deterioration, sometimes with fatal outcome. Burkholderia multivorans and Burkholderia cenocepacia are the species responsible for most of the infections of cystic fibrosis patients. Lipopolysaccharide endotoxins (LPSs) are among the foremost factors of pathogenesis of Gram-negative infection and, in particular, lipid A is the endotoxic portion of LPS responsible for eliciting host innate immune response. In this work, the complete primary structure of the lipid A from B. multivorans C1576 has been defined and, further, its pro-inflammatory activity in a cystic fibrosis airways model is shown. The structure of B. multivorans lipid A was attained by chemical, mass spectrometry and nuclear magnetic resonance analyses whereas its biological activity was assessed on the intestinal epithelial cell line CACO-2 cells, on the airway epithelial IB3-1 cells, carrying the ΔF508/W1282X CFTR mutation and on an ex vivo model of culture explants of nasal polyps.

Topics: Bronchi; Burkholderia; Caco-2 Cells; Cystic Fibrosis; Enzyme-Linked Immunosorbent Assay; Host-Pathogen Interactions; Humans; Interleukin-8; Lipid A; Magnetic Resonance Spectroscopy; Microscopy, Confocal; Nasal Polyps; Spectrometry, Mass, Electrospray Ionization; Tumor Necrosis Factor-alpha

The nasal epithelium is the first barrier encountered by airborne allergens and is an active participant in airway inflammation. Fungi have been increasingly recognized as important pathogens in sinusitis and airway diseases. The aim of the study was to evaluate fungal protease activity during cytokine production in nasal polyp epithelial cells and to determine the expression of Toll-like receptor (TLR) mRNA by fungi.. Nasal polyp epithelial cells were obtained from patients and stimulated with Alternaria and Aspergillus. Interleukin-8 (IL-8) and granulocyte macrophage colony-stimulating factor (GM-CSF) were measured to determine the activation of epithelial cells. Reverse transcriptase polymerase chain reaction for the TLR mRNA expression of the nasal epithelial cells was performed. Cytokine production was inhibited with protease inhibitors and anti-human TLR antibodies.. The fungi enhanced the production of IL-8 and GM-CSF from nasal epithelial cells. When nasal epithelial cells were activated by the fungi, TLR2, TLR3 and TLR4 mRNAs were more strongly expressed than in the nonactivated cells. Cytokine production was inhibited by protease inhibitors and anti-human TLR4 antibodies.. The results of this study showed that fungi interacted with nasal epithelial cells and enhanced the production of cytokines and TLR mRNA expression. The cytokine production was related to the protease in fungi and TLR4.

Topics: Air Microbiology; Alternaria; Aspergillus; Cells, Cultured; Epithelial Cells; Fungal Proteins; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Interleukin-8; Nasal Polyps; Peptide Hydrolases; Toll-Like Receptors

Biologically active vitamin D3 (VD) derivatives possess modulatory activities on immunological and inflammatory responses which can be reflected by altered levels of pro-inflammatory chemokines. Nasal polyposis (NP), defined as a chronic inflammatory process of upper respiratory system, could be influenced by VD derivatives. The purpose of this study was to investigate the influence of 1alpha,25-dihydroxyvitamin D3 (calcitriol) and 1alpha,24(R)-dihydroxyvitamin D3 (tacalcitol) on the secretion of IL-6 and IL-8 by fibroblasts derived from NP.. The study involved 12 fibroblast cultures derived from NP samples obtained from surgically treated patients. Measurements were performed on the polyp cells after the 6-9 passages. Culture stimulation involved treatment with tacalcitol and calcitriol at a defined strength (from 10(-7)M to 10(-4)M). IL-6 and IL-8 concentrations were estimated with ELISA.. Treatment with calcitriol or tacalcitol inhibits the synthesis of both IL-6 and IL-8 compared to the control group. The dose dependence of this effect has been confirmed. VD derivatives influence was marked at higher concentrations. Significant interleukin decrease was observed at 10(-5) and 10(-4) for calcitriol and 10-4 in the case of tacalcitol.. The present study demonstrates that calcitriol and tacalcitol are capable of affecting pro-inflammatory cytokine (IL-6 and IL-8) levels in NP cultures. Our data imply a potential therapeutical application of topical VD derivates in NP and warrant further investigation.

Topics: Calcitriol; Cells, Cultured; Dihydroxycholecalciferols; Fibroblasts; Gene Expression; Humans; Interleukin-6; Interleukin-8; Nasal Polyps; Vitamin D

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

Human nasal epithelial cells (hNECs) are the first line of immune defense and are able to produce mediators that recruit, activate and prolong survival of immune cells, among which IL-8 takes an important place. Studies on IL-8 and effects of dexamethasone on hNECs have been hampered by methodological shortcomings. The purpose of the study is to investigate the contribution of freshly isolated hNECs to IL-8 production in chronic rhinosinusitis with nasal polyps (CRSwithNP). Secondly, the effects of dexamethasone treatment on hNEC apoptosis and IL-8 production are investigated.. hNECs were freshly isolated from nasal polyp tissue and healthy inferior turbinate of NP patients (n=12) and from inferior turbinates of healthy donors (n=19) by protease treatment and two negative selection procedures. hNECs were incubated with IL-1β (10ng/ml), TNFα (10ng/ml) or dexamethasone (10, 100 and 1000 Amicrog/ml). After 24h, IL-8 levels were determined in the supernatants by ELISA. Finally, hNECs were incubated with increasing doses of dexamethasone and stained with trypan-blue and annexin-FITC/PI to study apoptosis.. hNECs isolated from nasal turbinates of healthy and NP patients and polyp tissue from NP patients produced similar levels of IL-8. IL-1β induced higher levels of IL-8 production in all types of hNECs without differences between control and NP tissue. Dexamethasone induced apoptosis of hNECs concomitant with abrogation of IL-8 production by hNECs.. IL-8 production by human nasal epithelial cells does not differ between NP and healthy tissue under baseline nor stimulatory conditions. Dexamethasone induces apoptosis of hNECs and abrogates IL-8 production.

Topics: Apoptosis; Cell Culture Techniques; Cells, Cultured; Dexamethasone; Dose-Response Relationship, Immunologic; Epithelial Cells; Glucocorticoids; Humans; Incubators; Interleukin-8; Nasal Mucosa; Nasal Polyps; Tumor Necrosis Factor-alpha

To investigate the expression of CD4, CD69, CD34, RANTES, IL-5 and IL-8 in nasal polyp tissues, and study their roles in the formation of nasal polyp.. The expression of CD4, CD69, CD34, RANTES, IL-5 and IL-8 were detected by immunohistochemical method and image analysis in 34 cases of nasal polyps and 30 cases of nasal concha mucosa (LNT).. The positive rate of glandular organ hyperplasia, formation of beaker cell, fiber hyperplasia, interstitial edema and infiltration of lymphocyte and eosinophilic granulocyte in nasal polyps were significantly higher than those in nasal concha mucosa (P<0.01). The cell density (piece/mm2) of CD4+, CD69+, IL-5, IL-8, RANTES in 34 nasal polyps was significantly higher than those in nasal concha mucosa (P<0.05). Marked positive correlations were found between expression of CD4, CD69 and RANTES, IL-5 and IL-8 (P<0.05, P<0.01 and P<0.05), expression of IL-5 and RANTES and infiltration level of eosinophilic granulocyte (P<0.05 and P<0.01), and expression of IL-8 and vaso formation on nasal polyps tissue (P<0.01).. T lymphocytes and correlated cytokines participate in the immunopathogenesis of nasal polyps; IL-5 and RANTES can prompt the infiltration, the aggregation and the activation of eosinophilic granulocytes; IL-8 can promote the vaso formation in nasal polyps.

Topics: Chemokine CCL5; Female; Granulocytes; Humans; Interleukin-5; Interleukin-8; Lymphocyte Activation; Lymphocyte Count; Male; Middle Aged; Nasal Mucosa; Nasal Polyps; T-Lymphocytes

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

Nasal polyps (NP), a subgroup of chronic rhinosinusitis, are characterized by interleukin 5 (IL-5) mediated infiltration of eosinophils in sinus mucosa, leading to pseudostratified ciliated columnar epithelium, thickening of the epithelial basement membrane and tissue edema. Matrix metalloproteinases (MMP) constitute a large group of Zn2+ dependent endopeptidases with the ability to degrade extracellular matrix and are possibly responsible for the development of tissue edema in chronic sinusitis.. The aim of this study was to determine the expression of MMP-2, MMP-9 and tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) mRNA and to locate the distribution of MMP-2, MMP-9 and TIMP-1 by immunohistochemistry in ethmoid sinus mucosa in NP. Furthermore the correlation between IL-5 or IL-8 and MMP-2, MMP-9 or TIMP-1 is examined.. Nasal polyps of 33 patients and 18 specimens of inferior turbinate mucosa were examined by real time RT-PCR for MMP-2, MMP-9, TIMP-1, IL-5 and IL-8 mRNA expression. Immunohistochemical labeling for MMP-2, MMP-9 and TIMP-1 was performed.. Differences between both locations were detectable for MMP-9 (P < 0.001) and IL-5 (P=0.003) but not for MMP-2 (P=0.278), TIMP-1 (P=0.515) and IL-8 (P=0.386). Correlation was detected only between TIMP-1 and IL-5 (r=0.422, P =0.014). Cytoplasmic staining of MMP-2 was present in the apical part of the ciliated cells, submucosal glands and in smooth muscle cells. Matrix metalloproteinase-9 was expressed in surface epithelium, in seromucous glands and in polymorphonuclear cells.. Expression of MMP-9 and IL-5 mRNA are associated with NP. The correlation between IL-5 and TIMP-1 indicates the role of TIMP-1 in maintaining the homeostasis in NP.

Topics: Adult; Aged; Female; Gene Expression; Humans; Immunohistochemistry; Interleukin-5; Interleukin-8; Male; Matrix Metalloproteinase 2; Matrix Metalloproteinase 9; Middle Aged; Nasal Polyps; Tissue Distribution; Tissue Inhibitor of Metalloproteinase-1

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

Second-generation antihistamines are H(1) receptor antagonists and may have additional anti-inflammatory effects.. The aims of the study were to evaluate the effect of desloratadine (DL) on cytokine secretion by epithelial cells from both nasal mucosa (NM) and polyps (NP), and on eosinophil survival primed by epithelial cell secretions.. Epithelial cells were cultured and stimulated with fetal bovine serum (FBS), IL-1beta or TNF-alpha with and without DL for 24 h. Culture supernatant cytokines concentration were measured by ELISA. Peripheral blood eosinophils were incubated with human epithelial cell conditioned media (HECM) and DL. Eosinophil survival was assessed by Trypan blue dye exclusion. Results are expressed as mean+/-SEM of cytokine concentration (pg/mL) or eosinophil survival index (%).. FBS increased granulocyte-macrophage colony-stimulating factor (GM-CSF), vascular endothelial growth factor (VEGF), IL-6, IL-8, and TGF-beta(1) secretion in epithelial cell cultures from both NM and NP. Only GM-CSF secretion was significantly (P<0.05) inhibited by a dose-response of DL compared with positive controls, in both NM (10(-5) m: 125+/-36 pg/mL, 10(-6) m: 95+/-22 pg/mL vs. control: 256+/-91 pg/mL, n=6) and NP (10(-5) m: 80+/-29 pg/mL, 10(-6) m: 109+/-45 pg/mL vs. control: 333+/-212 pg/mL, n=6). DL also showed an inhibitory effect on HECM-induced eosinophil survival from both NM and NP. At 72 h, DL significantly (P<0.01) inhibited eosinophil survival induced by HECM from NM (10(-5) m: 19.9+/-5.5%, n=9; 10(-6) m: 28.7+/-7.7%, n=9) and NP (10(-5) m: 6.2+/-2.8%, n=11) compared with HECM alone (NM: 42.1+/-7.3%; NP: 45.3+/-8.1%).. The inhibitory effects of DL on epithelial cell GM-CSF secretion and on eosinophil survival induced by epithelial cell secretions, suggest that this H(1) antagonist may regulate eosinophil inflammation in upper airways.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Case-Control Studies; Cell Survival; Cells, Cultured; Chemokine CCL11; Chemokine CCL5; Chemokines, CC; Enzyme-Linked Immunosorbent Assay; Eosinophils; Epithelial Cells; Female; Granulocyte-Macrophage Colony-Stimulating Factor; Histamine H1 Antagonists; Humans; Interleukin-5; Interleukin-6; Interleukin-8; Loratadine; Male; Middle Aged; Nasal Mucosa; Nasal Polyps; Statistics, Nonparametric; Transforming Growth Factor beta; Vascular Endothelial Growth Factor A

To study the concentration and expression of IL-4, IL-5, IL-6, IL-8 in human nasal polyps tissues, and to explore the relationship between cytokines and the formative mechanism of nasal polyps.. The concentration and expression of IL-4, IL-5, IL-6, IL-8 in 54 cases with nasal polyps were determined by enzyme linked immunosorbent assay (ELISA) and immunohistochemistry, and the middle turbinate mucosa from 22 patients with the deviation of nasal septum were used as the control.. The concentration of IL-5, IL-8 in nasal polyps tissues were significantly higher than that in normal middle turbinate mucosa (P < 0.01). There was no significant difference between the concentration of IL-6 in nasal polyps and that in normal control group (P > 0.05). No significant difference was found between the concentration of IL-4 in nasal polyps tissues with negative allergic skin test and that in control group, but the concentration of IL-4 was significantly higher in nasal polyps tissues with positive allergic skin test than in control group (P < 0.05). IL-4 was mostly found in lymphocytes and plasma cells in nasal polyps and IL-5 in eosinophils and lymphocytes. IL-6 and IL-8 were mostly found in nasal polyps epithelium and inflammatory cells, as was no difference between positive allergic skin test and negative one.. IL-5 and IL-8 are the key cytokines in the formation of nasal polyps, and IL-4 is a key cytokine in nasal polyps with positive allergic skin test. IL-6 isn't an important cytokine in the formative mechanism of nasal polyps.

Topics: Adolescent; Adult; Case-Control Studies; Female; Humans; Interleukin-4; Interleukin-5; Interleukin-6; Interleukin-8; Male; Middle Aged; Nasal Polyps; Young Adult

Proteases in fungi interact with nasal epithelial cells and enhance the production of inflammatory cytokines in vitro. These cytokines induced the migration of eosinophils and neutrophils. Protease-activated receptors (PARs) might also play a role in the process of epithelial cell activation.. The nasal epithelium is the first barrier encountered by airborne allergens and an active participant in airway inflammation. Fungi have been increasingly recognized as important pathogens in sinusitis and consist of several allergenic proteins.. Nasal polyp epithelial cells were obtained from patients and stimulated with Alternaria, Aspergillus, and Cladosporium. Interleukin-8 (IL-8), granulocyte-macrophage colony stimulating factor (GM-CSF), and regulated on activation normal T expressed and secreted (RANTES) were measured to determine the activation of epithelial cells. Reverse transcriptase-polymerase chain reaction test (RT-PCR) for PAR mRNA expression in nasal epithelial cells was performed. Eosinophil and neutrophil migration was induced with nasal polyp epithelial cells conditioned media (HPECM).. Fungi enhanced the production of chemical mediators from nasal epithelial cells. When nasal epithelial cells were activated with fungi, PAR2 and PAR3 mRNAs were more strongly expressed than in nonactivated cells. Eosinophil migration was induced by RANTES and eotaxin, and neutrophil migration was induced by IL-8 in HPECM.

Topics: Allergens; Cell Migration Inhibition; Cell Movement; Cells, Cultured; Chemokine CCL11; Chemokine CCL5; Chemokines, CC; Culture Media, Conditioned; Eosinophils; Epithelial Cells; Fungi; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Interleukin-8; Nasal Polyps; Neutrophils; Receptors, Proteinase-Activated; RNA, Messenger

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 dysfunction of the mucosal interface of the upper respiratory tract in cystic fibrosis (CF) patients is clinically visible by the development of nasal polyps (NP) at a young age. Innate defence markers and inflammatory mediators in NP from patients with CF were compared with non-cystic fibrosis nasal polyps (non-CF-NP) to determine a possible different immunological background in macroscopically similar tissue.. Surgical samples were obtained from patients with non-CF-NP, cystic fibrosis patients with nasal polyps (CF-NP) and control patients (CO). With real time PCR, the mRNA expression of human beta defensins (HBD) 2 and 3, toll-like receptors (TLR) 2 and 4 and the macrophage mannose receptor (MMR) were measured. On homogenates of the surgical samples eotaxin, myeloperoxidase (MPO), IL-5 and IL-8 protein content was measured using commercial ELISA kits; IgE and eosinophilic cationic protein (ECP) were measured by the Unicap system.. In CF-NP we found a statistically significant higher mRNA expression of HBD 2 compared with non-CF-NP and CO and of TLR 2 compared with non-CF-NP. In the non-CF-NP group, MMR mRNA expression was significantly elevated compared with CO and CF-NP. For TLR 4 mRNA expression no statistically significant differences were found between groups. IL-5 was below detection level in all CO and CF-NP, but was measurable in 80% of the non-CF-NP. MPO and IL-8 concentrations were significantly higher in CF-NP compared with CO and non-CF-NP, whereas ECP, eotaxin and IgE were significantly higher in the non-CF-NP group.. We here demonstrate that CF-NP and non-CF-NP not only differ in terms of inflammatory mediator profile, but also in terms of innate markers.

Topics: Anti-Infective Agents; beta-Defensins; Biomarkers; Cystic Fibrosis; Humans; Inflammation Mediators; Interleukin-5; Interleukin-8; Lectins, C-Type; Macrophages; Mannose Receptor; Mannose-Binding Lectins; Membrane Glycoproteins; Nasal Polyps; Peroxidase; Receptors, Cell Surface; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Toll-Like Receptor 2; Toll-Like Receptor 4; Toll-Like Receptors

Airway epithelial cells have a major role in initiating inflammation in response to bacterial pathogens. Through the immediate induction of CXCL8 and cytokine expression, polymorphonuclear cells are mobilized and activated to eradicate the infecting organisms. However, the influx of polymorphonuclear cells and the effects of their toxic exoproducts impede respiratory function. We postulated that respiratory epithelial cells must also participate in the regulation of their own proinflammatory signaling. Both Staphylococcus aureus and Pseudomonas aeruginosa were found to potently activate IL-6 expression immediately upon contact with epithelial cells, and by 1 h induced TNF-alpha converting enzyme (TACE) transcription. By 4 h of bacterial exposure, TACE colocalized with IL-6Ralpha on the apical surface of airway cells, and by 24 h, soluble IL-6Ralpha accumulated in the cell culture supernatant. Epithelial IL-6 and soluble IL-6Ralpha were shown to participate in trans-signaling, interacting with membrane-associated gp130 to activate CCL-2 expression and inhibit additional CXCL8 production. Thus, bacteria are physiological activators of TACE expression, which provides a mechanism to regulate inflammatory signaling that is initiated by airway epithelial cells.

Topics: ADAM Proteins; ADAM17 Protein; Bronchi; Cell Line; Chemokine CCL2; Enzyme Activation; Humans; Inflammation Mediators; Interleukin-6; Interleukin-8; Metalloendopeptidases; Nasal Polyps; Pseudomonas aeruginosa; Receptors, Interleukin-6; Respiratory Mucosa; Signal Transduction; Solubility; Staphylococcus aureus; Substrate Specificity; Tumor Necrosis Factor-alpha

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

Decreased connexin 43 (Cx43) expression as a result of application of lipopolysaccharide (LPS) may limit the diffusion of intercellular signaling molecules that is essential to the coordinated function of neighboring cells. Therefore, it may be related to a ciliary beating defect in nasal epithelial cells and result in accumulation of harmful substances.Gap junction intercellular communication (GJIC) is altered during inflammation in tracheal epithelial cells. Thus, we aimed to investigate whether LPS affects the expression of Cx43, the elementary protein composing the gap junction of nasal epithelial cells, in vitro.LPS (Pseudomonas aeruginosa serotype 10) was applied to epithelial cells obtained from nasal polyp for 24 h in vitro. As an inflammatory indicator, IL-8 secretion was measured using a commercially available ELISA kit. Cx43 protein was detected semi-quantitatively using Western blotting. The nasal epithelial cells constitutively secreted IL-8 at a concentration of 0.45+/-0.03 ng/microg protein. In the presence of 10(-2) mg/ml LPS, the concentration of IL-8 was significantly increased to 0.55+/-0.05 ng/microg protein (n=8). Expression of the Cx43:beta-actin ratio decreased in a time- and dose-dependent fashion (10(-3)-10(-1) mg/ml LPS).

Topics: Adolescent; Adult; Aged; Cell Culture Techniques; Connexin 43; Epithelial Cells; Female; Humans; Interleukin-8; Lipopolysaccharides; Male; Middle Aged; Nasal Mucosa; Nasal Polyps; Pseudomonas aeruginosa

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 study the expression of IL-8 and IL-3 in human nasal polyp and polyposis.. Using immunohistochemical methods detect the expressions of IL-8 and IL-3 in 52 cases of middle turbinate,nasal polyp and nasal polyposis,and calculating the positive area percentages in each group. All datas were analysed with SNK test, Kruskal-Wallis test and Mann-Whitney test.. There were very significant differences between either two groups about the expressions of IL-8. There were significant differences between either two groups about the expressions of IL-3.. IL-8 is not only one of the causes in nasal polyp, but also plays an important role in the recurrence of nasal polyposis. The expression of IL-8 in polyp is different from that in polyposis, it indicate that polyp and polyposis have different mechanism. IL-3 is important for the accumulation of eosinophil and is a key factor in the mechanism of polyposis.

Topics: Adolescent; Adult; Aged; Child; Eosinophils; Female; Humans; Interleukin-3; Interleukin-8; Male; Middle Aged; Nasal Mucosa; Nasal Polyps

Increased mucin expression is a feature of nasal polyposis. Corticosteroids reduce polyp size and symptoms, but their effect on mucin production remains unknown. In this study, the effects of intranasal corticosteroids on MUC5AC mucin expression, nasal resistance, eosinophil and neutrophil infiltration, epidermal growth factor receptor (EGFR), interleukin (IL)-8, and tumour necrosis factor (TNF)-alpha expression was assessed in nasal polyps. In nine subjects, one nasal polyp was removed surgically before treatment and another was removed after 8 weeks of intranasal fluticasone (400 microg.day(-1)). Tissues were processed for in situ hybridisation and immunohistochemical staining. Described effects of fluticasone on nasal polyps (reduction in nasal resistance and in eosinophil infiltration) were evaluated. Morphometric analysis was performed to assess the effect of fluticasone on epithelial-, MUC5AC-, EGFR- and IL-8-stained areas, TNF-alpha-stained cells, and neutrophil numbers. Treatment with fluticasone decreased nasal resistance and intra-epithelial eosinophils. The MUC5AC-stained area in the epithelium was unchanged by treatment; MUC5AC mRNA expression was unaffected by treatment. EGFR-stained area, intra-epithelial neutrophil numbers, IL-8 and TNF-alpha expression were also unchanged by therapy. Intranasal fluticasone was effective in decreasing nasal airflow resistance and intra-epithelial eosinophils but had no effect on mucin or epidermal growth factor receptor expression or on neutrophil recruitment.

Topics: Administration, Intranasal; Adrenal Cortex Hormones; Androstadienes; Eosinophils; ErbB Receptors; Fluticasone; Humans; Interleukin-8; Mucin 5AC; Mucins; Nasal Polyps; Neutrophil Infiltration; Tumor Necrosis Factor-alpha

Accumulating data show that fibroblasts are important regulators in the development and maintenance of allergic airway inflammation. However, most studies so far have used individual recombinant cytokines in high concentrations, unlikely to be found in vivo. We aimed to investigate how cytokines produced by peripheral blood mononuclear cells (PBMC) affect fibroblast functions. Primary airway fibroblasts where incubated with allergen-stimulated or non-stimulated PBMC supernatants from allergic patients. The levels of cytokines in PBMC supernatants were measured and the expression of CD54, CD40 and CD106 as well as the production of eotaxin, interleukin (IL)-6 and IL-8 were assessed in fibroblasts. Although the levels of single cytokines measured in PBMC supernatants were low, a significant up-regulation of the surface molecules as well as of IL-6 and IL-8 production was found in fibroblasts cultured with allergen-stimulated PBMC supernatants as compared to non-stimulated, while the increase in eotaxin production was not significant. The evaluation of correlations between cytokines produced by PBMC and effects seen on fibroblasts did not indicate a crucial role for any single cytokine. Furthermore, the addition of comparably low concentrations of recombinant interferon (rIFN)-gamma or recombinant tumour necrosis factor (rTNF)-alpha did not induce the same effects as PBMC supernatants, the only exception being TNF-alpha as a direct inducer of CD54 expression. Our results show that synergistic mechanisms has a more important role than single mediators, highlighting important differences between in vitro experiments, where effects of individual mediators are studied, versus the actual situation in vivo.

Topics: Cells, Cultured; Cytokines; Fibroblasts; Humans; Hypersensitivity; Intercellular Adhesion Molecule-1; Interferon-gamma; Interleukin-13; Interleukin-6; Interleukin-8; Leukocytes, Mononuclear; Lymphocyte Activation; Nasal Polyps; Tumor Necrosis Factor-alpha; Vascular Cell Adhesion Molecule-1

The influence of macrolide antibiotics, roxithromycin (RXM) and josamycin (JM) on inflammatory cytokine production from human nasal polyp fibroblasts (NPFs) was examined using an in vitro cell culture technique. Addition of RXM at a concentration of 10.0 microg/ml to cell cultures suppressed both IL-6 and RANTES (but not IL-8) production in response to stimulation with 25.0 ng/ml tumor necrosis factor (TNF)-alpha. However, JM could not suppress IL-6, IL-8 and RANTES production from NPFs induced by TNF-alpha stimulation in vitro, even when added to cell cultures at a concentration of 20.0 microgram/ml. In the second part of the study, we examined the influence of RXM on cytokine mRNA expression in NPFs. Addition of RXM at a concentration of 10.0 mg/ml to cell cultures caused reduction of the mRNA expressions of both IL-6 and RANTES, which were enhanced by TNF-alpha stimulation in vitro.

Topics: Anti-Bacterial Agents; Cell Culture Techniques; Chemokine CCL5; Cytokines; Fibroblasts; Humans; Interleukin-6; Interleukin-8; Josamycin; Nasal Polyps; RNA, Messenger; Roxithromycin; Tumor Necrosis Factor-alpha

Eosinophil infiltration into an inflammatory site is a characteristic histological finding in patients with chronic rhinosinusitis and nasal polyps. Most of the eosinophils in chronic rhinosinusitis are activated in the nasal cavity, but the exact activation mechanism of eosinophils is unknown. The study was designed to investigate the effect of human nasal epithelial cells on the activation of eosinophils.. Peripheral blood eosinophils were isolated from healthy volunteers and incubated in human nasal polyp epithelial cell conditioned media (HPECM). Superoxide production and eosinophil-derived neurotoxin were measured to determine eosinophils activation. HPECMs were assayed by ELISAs for interleukin-8 (IL-8), granulocyte-macrophage colony stimulating factor (GM-CSF), eotaxin, and regulated on activation normal T expressed and secreted (RANTES). To identify the chemical mediators involved in the activation of eosinophils.. HPECM (n = 7) contained 31.48 ng/mL interleukin-8, 533.43 pg/mL GM-CSF, 5.90 pg/mL eotaxin, and 11.06 pg/mL RANTES. Eosinophils were activated by HPECM and inhibited only by anti-GM-CSF antibody, not by the other chemical mediators.. The results suggest that eosinophils in nasal secretions are activated by GM-CSF, which is produced by nasal epithelial cells.

Topics: Cells, Cultured; Chemokine CCL11; Chemokine CCL5; Chemokines, CC; Culture Media, Conditioned; Eosinophil-Derived Neurotoxin; Eosinophils; Epithelial Cells; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Interleukin-8; Nasal Polyps; Ribonucleases; Superoxides

Topics: Anti-Bacterial Agents; Chemokine CCL5; Depression, Chemical; Dose-Response Relationship, Drug; Fibroblasts; Humans; Inflammation Mediators; Interleukin-6; Interleukin-8; Nasal Mucosa; Nasal Polyps; Roxithromycin

Nasal polyps frequently appear in patients with cystic fibrosis (CF). The aims of this study were to focus on what problems (symptoms, endoscopic findings, and laboratory correlates) nasal polyps cause the CF patient, and how these correlate to the total health situation of this patient group.. The clinical histories, endoscopic investigations of the nasal cavity, and analyses of nasal lavage fluid of 44 patients with CF complicated with nasal polyposis have been compared with those of 67 CF control subjects. The patients were examined at annual control examinations (with pulmonary tests, working capacity, liver tests, and bacterial and blood tests) from 1995 to 1996 at Stockholm Cystic Fibrosis Center, Huddinge University Hospital. All patients were > 2 years of age. The endoscopic findings were related to the actual pulmonary function, inflammatory blood parameters, colonizing pathogens, antibodies (Staphylococcus aureus and Pseudomonas aeruginosa), and genotype.. The patients with nasal polyps differed with respect to chronic colonization of P aeruginosa in sputum samples and had a higher occurrence of serum antibodies against the same species. The two groups did not differ in pulmonary functions, inflammatory parameters, or genotype. The polyps found were mainly small (within the meatus media) and gave no significant increase in ongoing clinical symptoms such as rhinorrhea, nasal obstruction, or hyposmia. Neither was any significantly marked finding concerning the nose (mucosal swellings, secretion, etc.) made in the polyp patients. The patients with CF scored slightly lower in a smell identification test in comparison with the healthy control group. The nasal lavage fluid was analyzed (in 93 of the 111 patients) for the occurrence of P aeruginosa (by polymerase-chain reaction [PCR]), interleukin [IL]-5, IL-8, and lysozyme. The lysozyme and IL-8 content was equal in the two CF groups but increased in comparison with the healthy control group. P aeruginosa was not detected with PCR in any nasal lavage fluid. No measurable levels of IL-5 in the nasal lavage were found.. There was a higher frequency of chronic colonization of P aeruginosa in the lower respiratory tract in patients with nasal polyps. Otherwise, nonsevere nasal polyposis was not an indicator of lower respiratory tract morbidity in CF patients.

Topics: Adolescent; Adult; Antibodies, Bacterial; Child; Child, Preschool; Cystic Fibrosis; Endoscopy; Female; Humans; Interleukin-5; Interleukin-8; Male; Muramidase; Nasal Lavage Fluid; Nasal Polyps; Pseudomonas aeruginosa; Pseudomonas Infections; Respiratory Tract Infections; Risk Factors; Staphylococcal Infections

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

To explore the role of interleukin-6,8 in nasal polyp formation and to search into the effect of allergy in the pathogenesis of nasal polyps (NP).. The expression and significance of interleukin-6,8 were studied in 36 nasal polyps and 36 serum samples of NP patients by radioimmunoassay.. The mean value of IL-6 and IL-8 was (2.7658 +/- 0.3797) ng/L and (4.1877 +/- 0.1758) ng/L in all nasal polyp tissue homogenates. As compared with serum of NP patients, IL-6 and IL-8 were over expressed in nasal polyp tissue homogenates. No relation was found between the expression of IL-6/IL-8 and patients' gender, age and clinical stage. The expression of IL-6 and IL-8 in patients' serum, cord blood and normal serum showed no significant difference.. IL-6 and IL-8 are strongly correlated with the formation of nasal polyp. Neither allergy nor infection play a role in the pathogenesis of nasal polyps.

Topics: Adolescent; Adult; Aged; Child; Female; Humans; Interleukin-6; Interleukin-8; Male; Middle Aged; Nasal Polyps

We investigated the effect of budesonide and nedocromil sodium on the secretion of IL-6 and IL-8 by cultured epithelial cells from healthy nasal mucosa and nasal polyps. Human epithelial cell conditioned media was generated with fetal calf serum (FCS) in the presence or absence of budesonide and/or nedocromil sodium. Budesonide inhibited FCS-induced IL-6 and IL-8 release in a dose-dependent manner. The IC25 (25% inhibitory concentration) of budesonide on IL-6 release was higher in nasal polyp than in nasal mucosa epithelial cells (34 nM vs. 200 pM). The IC25 of budesonide on IL-8 release was higher in nasal mucosa than in nasal polyps (145 pM vs. 4 pM). Nedocromil sodium caused a dose-related inhibitory effect on IL-8 release from nasal mucosa (IC25, 207 nM), while it only had a significant effect in nasal polyps at 10(-5) M. Nedocromil sodium had no effect on IL-6 release. The inhibitory effect of budesonide was higher than that of nedocromil sodium on both nasal polyps and nasal mucosa. Budesonide and nedocromil sodium may exert their anti-inflammatory action in the respiratory mucosa by modulating the secretion of IL-6 and IL-8. The different effect of budesonide and nedocromil sodium on IL-6 and IL-8 release may be explained by differences in the mechanisms which regulate the upregulation of these cytokines in inflammatory responses.

Topics: Adolescent; Adult; Aged; Anti-Inflammatory Agents; Budesonide; Cells, Cultured; Dose-Response Relationship, Drug; Enzyme-Linked Immunosorbent Assay; Female; Humans; Interleukin-6; Interleukin-8; Male; Microbial Sensitivity Tests; Middle Aged; Nasal Mucosa; Nasal Polyps; Nedocromil; Statistics, Nonparametric

To investigate the distribution and content of immunoreaction interleukin-8(IL-8) antigen in the human nasal polyps.. Twenty-two cases of nasal polyp and 10 cases of normal inferior turbinate were studied with immunohistochemical SABC method and ELISA method.. Immunohistochemical results demonstrated that IL-8 antigen staining occurred predominantly within inflammatory and epithelium cell plasm. The level of IL-8 in nasal polyp tissue was higher than in normal control specimens. The value in nasal polyp tissue was in 322.06-2091.41 pg/ml, but in normal with a range of 29.31-332.19 pg/ml. The results demonstrated a remarkable difference between the two groups (P < 0.001).. These data demonstrated the presence of IL-8 antigen in nasal polyps. Thus IL-8 likely plays a significant role in the pathogenesis of this disease process. There is an important significance of finding a potential treatment method.

Topics: Adolescent; Adult; Female; Humans; Interleukin-8; Male; Middle Aged; Nasal Polyps; Turbinates

Topics: Adult; Cell Adhesion Molecules; Cells, Cultured; Chemokine CCL2; Chemokine CCL5; Chemokines; Cytokines; Fibroblasts; Humans; Intercellular Adhesion Molecule-1; Interleukin-1; Interleukin-13; Interleukin-4; Interleukin-5; Interleukin-6; Interleukin-8; Nasal Mucosa; Nasal Polyps; Nose Neoplasms; Vascular Cell Adhesion Molecule-1

We have used the Stamper-Woodruff frozen-section assay (FSA) to characterize the integrin and activation steps involved in adhesion of peripheral blood eosinophils and neutrophils to nasal polyp endothelium (NPE). Eosinophil and neutrophil adhesion was significantly inhibited by monoclonal antibodies (mAbs) against CD18 (beta2) and CD11a-c. Eosinophil adhesion was also inhibited to a lesser extent by mAbs against CD29 (beta1), CD49d (alpha4), and vascular cell adhesion molecule-1. The involvement of integrins raised the possibility of an activation step being involved in the adhesion process. Although stimulation of the cells with granulocyte macrophage colony-stimulating factor (GM-CSF) before the assay failed to modulate adhesion, binding was inhibited by up to 50% by treatment of the leukocytes with azide. In addition, neutrophil adhesion was completely abrogated by pertussis toxin (PT) and inhibited by about 50% by the platelet-activating factor antagonist WEB 2086 and antibodies against interleukin (IL)-8 and the two IL-8 receptors IL8RA and IL8RB (C-X-CR1 and -CR2). In contrast, eosinophil adhesion was unaffected by PT, WEB 2086, or anti-IL8R mAbs. mAbs against CCR-3, IL-3, IL-5, and GM-CSF also had no effect. This study demonstrates that eosinophil and neutrophil adhesion to NPE in the FSA conforms to the multistep paradigm for leukocyte adhesion and can be used to model the molecular basis for adhesion to endothelium in the context of chronic inflammatory disease. Using this assay, we have observed significant differences in integrin usage between eosinophils and neutrophils and a striking difference in the mechanism of integrin activation. These differences could explain, in part, the preferential accumulation of eosinophils in diseases such as asthma.

Topics: CD11 Antigens; CD18 Antigens; Cell Adhesion; Endothelium; Eosinophils; Epithelium; Granulocyte-Macrophage Colony-Stimulating Factor; GTP-Binding Proteins; Humans; Inflammation; Integrins; Interleukin-3; Interleukin-5; Interleukin-8; Nasal Polyps; Neutrophils; Pertussis Toxin; Platelet Activating Factor; Receptors, CCR3; Receptors, Chemokine; Virulence Factors, Bordetella

Topics: Anti-Bacterial Agents; Cells, Cultured; Fibroblasts; Humans; Interleukin-8; Nasal Polyps; Roxithromycin

Although several studies have demonstrated that low-dose, long-term 14-member macrolides (erythromycin (EM), roxithromycin (RXM), clarithromycin (CAM)) are effective in the treatment of chronic airway diseases like chronic sinusitis and diffuse panbronchiolitis (DPB), the mechanism of action of these drugs is not yet clear. Both these airway diseases are associated with an increase in the proliferation of fibroblasts. Moreover, fibroblasts are also an important source of proinflammatory cytokines such as interleukin-8 (IL-8), that play an important role in the pathogenesis of nasal polyps. Therefore, using primary fibroblast lines derived from nasal polyps, we investigated the effect of RXM on the synthesis of IL-8 and proliferation of nasal polyp fibroblasts (NPF). These fibroblasts were either treated with lipopolysaccharide (LPS) and RXM for 24 h, or pre-incubated with RXM for 24 h and then treated with LPS and RXM for 24 h. The level of IL-8 mRNA in NPF was analysed by reverse transcriptase-polymerase chain (RT-PCR) and the level of IL-8 in culture supernatants was measured by ELISA. Next, the proliferative capacity of NPF after treatment with RXM was analysed by cell counting and 3H-thymidine uptake. RXM had no effect on LPS-induced IL-8 synthesis by NPF. On the other hand, RXM suppressed the proliferation of NPF in a dose-dependent manner. These findings suggest that, although RXM cannot directly inhibit the synthesis of IL-8, it probably reduces IL-8 production by inhibiting the proliferation of NPF.

Topics: Anti-Bacterial Agents; Cell Count; Cell Line; Dose-Response Relationship, Drug; Fibroblasts; Humans; Interleukin-8; Lipopolysaccharides; Nasal Polyps; Roxithromycin

Eosinophilia is a feature of nasal polyposis. The aim of this study was to determine the role of cytokines and allergen in maintaining the eosinophilic infiltrate in this condition. Polyp fragments from house dust mite (HDM)-sensitive atopic individuals and nonatopic individuals were cultured in the presence of HDM, or phytohaemagglutinin (PHA) or culture medium alone. Culture supernatants were assayed for interleukins (IL) 3, 5, and 8 and granulocyte macrophage colony stimulating factor (GM-CSF), and eosinophil survival enhancing activity (ESEA) in vitro. Significant ESEA was produced spontaneously. When polyp tissue from atopics, but not from nonatopics, was stimulated with allergen for 2 days there was a further increase in ESEA associated with a median 12 and fourfold increase in IL-8 and GM-CSF, respectively. This increased ESEA was markedly reduced with anti-GM-CSF and, to a lesser extent, anti-IL-8 blocking antibodies. When stimulated with PHA, polyp tissue from atopic subjects also produced increased ESEA, implicating possible T-cell involvement. This was associated with a small (twofold), but significant, increase in IL-8 and a less consistent increase in GM-CSF. However, anti-IL-8 or anti-GM-CSF blocking antibodies failed to reduce the ESEA in these supernatants, suggesting involvement of other mechanisms. This study suggests that in sensitized individuals, allergen may contribute to polyp eosinophilia by stimulating the production of granulocyte/macrophage colony stimulating factor and interleukin 8.

Topics: Allergens; Animals; Cell Survival; Dust; Eosinophilia; Eosinophils; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Hypersensitivity, Immediate; In Vitro Techniques; Interleukin-8; Mites; Nasal Polyps; Tumor Cells, Cultured

The cytokine interleukin 8 (IL-8) has been shown to be a potent mediator of leukocyte recruitment and neovascularization in inflammatory and neoplastic diseases. In this study we hypothesize that IL-8 produced in the nasal polyp microenvironment is responsible for the leukocyte recruitment seen in nasal polyposis. To test this hypothesis we evaluated nasal polyps for distribution and content of IL-8 antigen with immunohistochemical techniques and radioimmunoassay to determine tissue levels of IL-8. The immunohistochemical results demonstrated that IL-8 antigen staining occurred predominantly within inflammatory cells and epithelium. IL-8 was detected in all nasal polyp tissue homogenates (a mean value of 1767 +/- 1633 pg/mg total protein (TP) with a range of 134 to 3668 pg/mg TP vs control specimens with a mean value of 77 pg/mg TP with a range of 0.09 to 255 pg/mg TP). These data demonstrate the presence and distribution and levels of IL-8 antigen in nasal polyps in vivo, supporting our hypothesis that local production of IL-8 could be an important factor in the sustained recruitment of leukocytes in nasal polyposis. Thus IL-8 likely plays a significant role in the pathogenesis of this disease process and therefore is a potential target for therapeutic intervention.

Topics: Adult; Aged; Antigens; Coloring Agents; Eosinophils; Epithelial Cells; Female; Gene Expression Regulation, Neoplastic; Humans; Immunoenzyme Techniques; Immunohistochemistry; Interleukin-8; Leukocytes; Male; Mast Cells; Middle Aged; Nasal Polyps; Neoplasm Proteins; Neovascularization, Pathologic; Plasma Cells; Radioimmunoassay

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