interleukin-8 and Otitis-Media-with-Effusion

To study the concentration of interleukin 8 (IL-8) in the middle ear fluid of children with acute otitis media and the association between IL-8 concentrations, aetiology of acute otitis media, and bacteriological sterilisation.. Middle ear fluid was obtained by tympanocentesis at enrollment (day 1) and on day 4-5 in 81 children aged 3-36 months with acute otitis media who received antibiotic treatment. IL-8 concentrations were measured by enzyme linked immunosorbent assay.. 101 samples were obtained on day 1 and 47 samples on day 4-5. 94 pathogens were isolated in 79 of 101 samples obtained on day 1: 56 Haemophilus influenzae, 35 Streptococcus pneumoniae, 2 Moraxella catarrhalis, and 1 Streptococcus pyogenes. Among 40 paired, initially culture positive samples, sterilisation was achieved on day 4-5 in 22 but not in 18 (13 H influenzae, 2 S pneumoniae, and 3 H influenzae and S pneumoniae concomitantly). IL-8 was detected in 96 of 101 and 46 of 47 samples obtained on days 1 and 4-5, respectively. Mean (SD) IL-8 concentration on day 1 was significantly higher in culture positive than in negative samples (12,636 (23,317) v 5,920 (7,080) pg/ml). In paired samples, IL-8 concentration fell in 12 of 22 ears in which sterilisation was achieved and in 9 of 21 ears with persistent or new infection. Mean (SD) IL-8 concentrations on day 4-5 were significantly higher in culture positive than in negative samples (15,420 (15,418) v 6,695 (5,092) pg/ml).. Higher IL-8 concentrations are found in culture positive middle ear fluid in acute otitis media. Bacterial eradication is associated with a fall in these concentrations.

Topics: Acute Disease; Anti-Bacterial Agents; Bacterial Infections; Child, Preschool; Exudates and Transudates; Humans; Infant; Interleukin-8; Otitis Media with Effusion

Bacteria infection and laryngopharyngeal reflux (LPR) were believed the important pathogenesis of chronic otitis media with effusion (COME). But no study researched the relationship between them on COME.. To confirm bacterial could arrive middle ear through LPR and produced acid metabolites to activate the pepsinogen of LPR causing COME.. Children (65) diagnosed COME with 122 middle ear effusions were included in COME group. Children (22) with congenital/acquired profound deafness with 22 middle ear lavage were included in CI group. Pepsin A concentration in the effusion and lavage fluid were measured. The DNA of the bacteria, IL-8 and TNF-α in the effusion were detected.. The average concentration of pepsin A in the effusions and lavage were 176.65 ± 242.09 and 19 ng/ml. Bacterial infection rates were 75.76% and 24.24% in the pepsin A(+) and pepsin A(-) patients. In the bacterial (+), the patients of pepsin A(+) was 4.33 times higher than those of pepsin A(-). TNF-α in pepsin A(+) was higher than that in pepsin A(-). TNF-α and IL-8 were higher in bacteria(+) than those of bacteria(-).. Bacterial infection and LPR might act in synergy in the pathogenesis of COME.. First time to propose LPR and bacterial infection might work synergistically to cause COME.

Topics: Adolescent; Bacterial Infections; Child; Child, Preschool; Chronic Disease; Cochlear Implants; Deafness; Ear, Middle; Female; Humans; Interleukin-8; Laryngopharyngeal Reflux; Male; Middle Ear Ventilation; Otitis Media with Effusion; Pepsin A; Prospective Studies; Tumor Necrosis Factor-alpha

Several cytokines and innate immune-associated molecules are present in middle ear effusions, but damage-associated molecular patterns (DAMPs) in middle ear effusion have not been studied. Therefore, we evaluated the role of heat shock proteins (Hsps) in the development of otitis media with effusion (OME).. Serous middle ear effusions from 22 pediatric patients who were diagnosed with OME and underwent ventilation tube insertion from June 2015 to March 2017 were evaluated in our study. The levels of Hsp 90, 70, 27, IL-8, and TNF-α in effusion fluids were evaluated by enzyme-linked immunosorbent assays. The associations between the levels of these molecules and the degree of tympanic membrane inflammation were statistically evaluated. Finally, the relationships among these molecules were also evaluated.. Hsp 70 and Hsp 27 were detected in all middle ear effusions, but Hsp 90 was detected in only five effusion fluid samples. IL-8 was also detected in all middle ear effusions, but TNF-α was detected in only four effusion fluid samples. When we compared the degree of tympanic membrane inflammation with the levels of Hsp 70, Hsp 27, and IL-8, which were detected in all effusion fluids, we could not find statistical significance. However, Hsp 70, Hsp 27, and IL-8 were significantly associated with each other (p < 0.05).. Hsp 70 and Hsp 27 were expressed in middle ear effusions. Furthermore, the levels of Hsp 70 and Hsp 27 were positively correlated with each other, and were also positively associated with the neutrophil chemoattractant, IL-8. Our findings suggested that Hsp 70 and Hsp 27 might be involved in the pathophysiology of pediatric OME.

Topics: Child; Child, Preschool; Enzyme-Linked Immunosorbent Assay; Female; Heat-Shock Proteins; HSP27 Heat-Shock Proteins; HSP70 Heat-Shock Proteins; Humans; Infant; Interleukin-8; Male; Middle Ear Ventilation; Molecular Chaperones; Otitis Media with Effusion; Tumor Necrosis Factor-alpha; Tympanic Membrane

Chronic otitis media with effusion (OME) is associated with irreversible changes in the middle ear, sometimes leading to hearing loss and abnormal language development in children. While the pathogenesis of OME is not fully understood, inflammatory and allergic factors are thought to be involved. The study aimed to investigate the role of cytokines in the local development of chronic OME, and assess differences in the cytokine profiles between atopic and non-atopic children. 84 atopic and non-atopic children with chronic OME (mean age of 6 years 7 months) were studied. Age-matched children with hypertrophy of the adenoids and Eustachian tube dysfunction served as the control group. The number of past acute otitis media (AOM) episodes, their age, and the type of effusion were recorded for all children. Pro-inflammatory cytokine concentrations (TNF-α, IL-1β, IL-6 and IL-8) were determined and the presence of pathogenic bacteria in the patients' effusions was examined. High concentrations of TNF-α, IL-1β, IL-6 and IL-8 were found in the effusions in all children with chronic OME, with the highest levels observed in the non-atopic group. The atopic group showed persistently high IL-1β levels, while in the non-atopic children, IL-1β and TNF-α levels positively correlated with the patient's age and the number of past AOM episodes. Pathogenic bacteria were more frequently isolated from effusions in non-atopic children. In both atopic and non-atopic children, pro-inflammatory cytokines are found at high concentrations. This argues in favor of instituting anti-inflammatory management for treating OME, regardless of atopy.

Topics: Case-Control Studies; Child; Child, Preschool; Chronic Disease; Ear, Middle; Female; Hearing Loss; Humans; Interleukin-1beta; Interleukin-6; Interleukin-8; Interleukins; Male; Otitis Media with Effusion; Tumor Necrosis Factor-alpha

Otitis media is characterized as an ongoing inflammation with accumulation of an effusion in the middle ear cleft. The molecular mechanisms underlying the pathogenesis, particularly the inflammatory response, remain largely unknown. We hypothesize that aspiration of gastric contents into the nasopharynx may be responsible for the initiation of the inflammatory process or aggravate a preexisting condition.. To investigate the correlation of gastric pepsin A with inflammatory cytokines, bacterial infection, and clinical outcomes.. Prospective study of 129 pediatric patients undergoing myringotomy with tube placement for otitis media at a tertiary care pediatric hospital.. Ear samples were tested for pepsin A; cytokines interleukin (IL)-6, IL-8, and tumor necrosis factor; and bacterial culture inoculation. Data were analyzed by descriptive statistics and regression analysis to identify risk factors for the presence of pepsin A and to correlate pepsin A levels with cytokine levels, infection status, and clinical outcomes.. Of the 129 patients, 199 ear samples were obtained; 82 samples (41%) and 64 patients (50%) were positive for pepsin A as measured by immunoassay. Pepsin A positivity correlated with age younger than 3.0 years (mean [SD], 2.3 [2.1] years in the positive group vs 3.3 [3.0] years in the negative group) and with all 3 cytokine levels (mean [SD] tumor necrosis factor, 29.5 [45.9] pg/mL in the positive group vs 13.2 [21.6] pg/mL in the negative group; IL-6, 6791.7 [9389.1] pg/mL in the positive group vs 2849.9 [4066.3] pg/mL in the negative group; and IL-8, 6828.2 [8122.3] pg/mL in the positive group vs 2925.1 [3364.5] pg/mL in the negative group [all P < .05]); however, logistic regression analysis showed that only IL-8 (odds ratio, 3.96; 95% CI, 1.3-12.0; P = .02) and age (odds ratio, 3.83; 95% CI, 1.2-12.7; P = .03) were significant independent variables. No statistically significant association was found with other parameters. Multiple linear regressions revealed that the levels of pepsin A were correlated with IL-8 levels (R2 = 0.248; P < .001) and the need for second or third tubes 6 to 12 months after the first (R2 = 0.102; P = .006). The presence of pepsin A in the middle ear was not associated with increased bacterial infection. Interleukin 8 was independent and significantly associated with both pepsin A levels and bacterial infection (R2 = 0.144 and 0.263, respectively; P = .001 for both).. Extraesophageal reflux as indicated by the presence of pepsin A is closely involved in the middle ear inflammatory process and may worsen the disease in some children; however, a proof of cause and effect between extraesophageal reflux and middle ear inflammation requires further investigation.

Topics: Child; Child, Preschool; Female; Gastroesophageal Reflux; Haemophilus influenzae; Humans; Infant; Interleukin-6; Interleukin-8; Male; Middle Ear Ventilation; Moraxella catarrhalis; Otitis Media with Effusion; Otitis Media, Suppurative; Pepsin A; Prospective Studies; Risk Factors; Streptococcus pneumoniae; Tumor Necrosis Factor-alpha

Otitis media (OM) is one of the most common childhood diseases. The relative contribution of complement activation in protection and pathogenesis during OM remains largely unknown. The purpose of this study was to investigate the beneficial and pathogenic contributions of complement activation in the middle ear of pediatric patients with recurrent acute otitis media (rAOM), and therefore to provide a rational approach to prevent sequelae of OM such as hearing loss.. Twenty children undergoing pressure equalization tube placement with or without adenoidectomy for rAOM were enrolled in the study. Bacterial cultures, enzyme-linked immunosorbent assay (ELISA) for complement components and cytokines and western blot for complement activation were performed on middle ear effusion (MEE) and serum samples. The levels of complement C3a, C5a and sC5-b9 in MEEs and serum samples were compared. The levels of these factors were also examined in regards to length of episode. Pearson's correlation coefficients were calculated on variables between C5a and IL-6 or IL-8. Complement gene expression in human middle ear epithelial (HMEE) cells induced by otopathogens was evaluated. Data were analyzed with Student's t test or the Mann-Whitney rank sum test. In all cases, a P value of <0.05 was set as the measure of significance.. Our data demonstrated that the complement classical/lectin, alternative and terminal pathways were activated in the middle ear of children with rAOM. Increased complement components of C3a, C5a and sC5-b9 in MEEs were detected in patients with the episode lasting more than six weeks. There was a strong correlation between C5a and IL-6 or IL-8 in the MEEs. Additionally, otopathogens induced enhanced gene expression of factor B and C3 in HMEE cells, which is beneficial for host defense against invading pathogens.. Our studies provided important new insights on how complement activation contributes to inflammatory process during rAOM. Knowledge of the activity of the complement pathway in patients with rAOM may stimulate the development of new strategies to prevent middle ear inflammatory tissue destruction by directing treatment to specific pathways within the complement cascade.

Topics: Acute Disease; Biomarkers; Blotting, Western; Child, Preschool; Cohort Studies; Complement Activation; Complement C3; Complement C5; Complement System Proteins; Enzyme-Linked Immunosorbent Assay; Female; Follow-Up Studies; Humans; Infant; Interleukin-6; Interleukin-8; Male; Middle Ear Ventilation; Otitis Media with Effusion; Polymerase Chain Reaction; Prospective Studies; Recurrence; Sensitivity and Specificity; Severity of Illness Index; Statistics, Nonparametric; Treatment Outcome

Vascular endothelial growth factor (VEGF) is a potent and critical inducer of angiogenesis and vascular permeability, and has been shown to play an important role in inflammatory events, together with hypoxia and inflammatory cytokines. VEGF messenger ribonucleic acid (mRNA) is expressed in the middle ear in an experimental animal model of otitis media with effusion (OME) and in patients with OME. However, the protein levels of VEGF in middle ear effusions (MEEs) are unknown and the role of VEGF in the pathogenesis of OME is unclear. The goals of this study were to determine the VEGF levels in MEEs and to investigate the role of VEGF in production of MEEs by comparing these levels with those of interleukin-8 (IL-8), endotoxin, and albumin.. Forty-six MEEs obtained from 33 children (24 boys, 9 girls) were used in the study. The mean age of the subjects was 6.3 years old (range, 1-12 years old). The patients underwent myringotomy and/or insertion of a ventilation tube for treatment of OME. After myringotomy, MEEs were collected with a Juhn Tym-Tap. The samples were divided into serous and mucoid types based on observation by the naked eye. After measuring the weight of the MEE, the sample was diluted with phosphate-buffered saline and frozen until use. The concentrations of VEGF and IL-8 in the MEEs were determined by enzyme-linked immunosorbent assays, endotoxin concentrations were measured by the Limulus Amebocyte Lysate test, and albumin levels were determined using an immunoturbidimetric assay.. VEGF, endotoxin, IL-8, and albumin were detected in 100%, 89%, 98%, and 100% of the 46 MEEs, respectively. The concentrations of VEGF, endotoxin, and IL-8 were significantly higher in mucoid MEEs than in serous MEEs (p<0.01), whereas there was no significant difference in the albumin concentration between mucoid and serous MEEs. The VEGF levels were positively correlated with those of endotoxin (R(2)=0.17, p<0.05) and albumin (R(2)=0.65, p<0.01) in mucoid MEEs, but not in serous MEEs.. Our results suggest that VEGF is produced in response to bacterial components such as endotoxin in the middle ear cavity and is associated with production of mucoid MEEs by increasing serum exudation and mucosal secretion.

Topics: Capillary Permeability; Child; Child, Preschool; Ear, Middle; Endotoxins; Exudates and Transudates; Female; Humans; Infant; Interleukin-8; Male; Middle Ear Ventilation; Otitis Media with Effusion; Serum Albumin; Vascular Endothelial Growth Factor A

We examined the effect of platelet-activating factor (PAF) on IL-8 production in cultured rat middle ear epithelial cells (RMECs), and the concentrations of cytokine, PAF, and PAF-acetylhydrolase (PAF-AH) were also examined in the PAF-induced experimental OME (otitis media with effusion) of rats.. Using an enzyme-linked immunospecific assay, we measured the levels of cytokines in the cultured RMECs and the PAF-induced OME of rats. The PAF was quantitated by the platelet-aggregating activity and the PAF-AH was measured by radioimmunoassay.. Both PAF and C-PAF, which is a stable analogue of PAF, significantly induced production of IL-8 in the RMECs in a dose-dependent manner. The PAF-induced IL-8 production was abolished by co-incubation with WEB2170, a specific PAF receptor antagonist. The concentrations of the cytokines and PAF in the PAF-induced OME of rats were higher on day 1 and the PAF and cytokine levels seemed to correspond well with the persistence of OME.. PAF may stimulate the local production of cytokines and may induce OME in the middle ear.

Topics: 1-Alkyl-2-acetylglycerophosphocholine Esterase; Animals; Cells, Cultured; Dose-Response Relationship, Drug; Epithelial Cells; In Vitro Techniques; Interleukin-8; Male; Otitis Media with Effusion; Platelet Activating Factor; Rats; Rats, Wistar

To elucidate the role of Moraxella catarrhalis lipooligosaccharide (LOS) in otitis media with effusion (OME), the effects of LOS on adhesion antigens of human monocytes were investigated. M. catarrhalis LOS selectively enhanced intercellular adhesion molecule 1 (ICAM-1 or CD54) expression on human monocytes by significantly increasing both the surface expression intensity and the percentage of ICAM-1(+) cells. ICAM-1 upregulation on human monocytes by the LOS required surface CD14, TLR4, NF-kappaB p65 and c-Jun N-terminal kinase (JNK) activity. Our study also revealed that the LOS-induced surface ICAM-1 expression was partially mediated through a TNF-alpha dependent autocrine mechanism and could be further augmented by lipopolysaccharide-binding protein in serum. In addition, M. catarrhalis LOS also stimulated human monocytes to produce pro-inflammatory cytokines in both TLR4- and CD14-dependent pathways. Our results also indicated that enhanced surface ICAM-1 expression on monocytes may hinder their adherence to the lung epithelial monolayer. Furthermore, the LOS-activated human monocytes secreted a significantly high level of IL-8, and could stimulate adjacent naïve monocytes to produce TNF-alpha which was partially mediated via membrane ICAM-1 and IL-8/IL-8RA. These results suggest that M. catarrhalis LOS could induce excessive middle ear inflammation through a cellular cross-talk mechanism during OME.

Topics: Acute-Phase Proteins; Carrier Proteins; Cell Communication; Cells, Cultured; Child; Humans; Intercellular Adhesion Molecule-1; Interleukin-8; JNK Mitogen-Activated Protein Kinases; Lipopolysaccharide Receptors; Lipopolysaccharides; Membrane Glycoproteins; Monocytes; Moraxella catarrhalis; Otitis Media with Effusion; Respiratory Mucosa; RNA, Small Interfering; Signal Transduction; Toll-Like Receptor 4; Transcription Factor RelA; Tumor Necrosis Factor-alpha

Chronic secretory otitis relates to the permanent presence of secretion in the middle ear for more than 3 months. The reason why applied therapy is often ineffective is that, for now, etiopathogenic molecular mechanisms responsible for the cause and the course of the secretory process in the mucus of the middle ear have not been precisely defined. Cytokines are the key mediators in middle ear inflammation with secretory otitis and regulating different inflammation states can add to the cause of the molecular processes that lead to hystopathological changes in mucus and submucus characteristically for the chronic state of secretory otitis. The aim of our work was to define the pro-inflammatory, immunoregulatory and allergy-associated cytokine levels in middle ear secretion samples of diseased children and to compare the defined values with the secretory process continuance in groups of patients who were diseased for more or less than 3 months. According to the results that have showed higher concentration of all ten examined cytokines in the secretion samples of the children who had secretory otitis for a longer time, it can be concluded that the disturbance expression regulation of the pro-inflammatory TNFalpha, TNFbeta, IL1beta, IFNgamma, IL-6 and IL-8, as well as immunoregulatory IL-2 and IL-10, and allergy associated cytokines IL-4 and IL-5 relating to the hyper production can add to the conversion of the inflammatory process to the chronic state, which has been maintained for longer than 3 months.

Topics: Adolescent; Child; Child, Preschool; Chronic Disease; Female; Humans; Infant; Interleukin-10; Interleukin-1beta; Interleukin-6; Interleukin-8; Lymphotoxin-alpha; Male; Mucous Membrane; Otitis Media with Effusion; Tumor Necrosis Factor-alpha

Alloiococcus otitidis has been found to be associated with otitis media with effusion. In this study we investigated whether TLR2 and collectins, surfactant protein A (SP-A) and mannose-binding lectin (MBL), interacted with A. otitidis. Both SP-A and MBL bound to A. otitidis in a Ca(2+)-dependent manner. A. otitidis induced IL-8 secretion from U937 cells and NF-kappaB activation in TLR2-transfected HEK293 cells. However, the cells transfected with the mutant TLR2(P681H) did not respond to A. otitidis. In addition, A. otitidis co-sedimented a recombinant soluble form of the extracellular TLR2 domain, indicating direct binding of the bacterium to TLR2. SP-A and MBL augmented the phagocytosis of A. otitidis by J774A.1 cells. The collectin-stimulated phagocytosis of A. otitidis was significantly attenuated when fucoidan and polyinosinic acid were co-incubated. Immunoblotting analysis revealed that MBL was present in the middle ear effusion from patients with otitis media. These results demonstrate that A. otitidis is a ligand for the collectins and TLR2, and that the collectins enhance the phagocytosis of A. otitidis by macrophages, suggesting important roles of the collectins and TLR2 in the innate immunity of the middle ear against A. otitidis infection.

Topics: Animals; Anti-Bacterial Agents; Gram-Positive Bacteria; Gram-Positive Bacterial Infections; Humans; Immunoblotting; Interleukin-8; Ligands; Mannose-Binding Lectin; Mice; NF-kappa B; Otitis Media with Effusion; Phagocytosis; Poly I; Polysaccharides; Rats; Signal Transduction; Toll-Like Receptor 2; Toll-Like Receptor 4; U937 Cells

This pilot study was designed to determine the serum cytokine profile of acute otitis media (AOM) due to Streptococcus pneumoniae and the impact of clarithromycin (Abbott Laboratories, Inc). Serum levels of interleukin-1 beta (IL-1 beta), tumor necrosis factor alpha (TNF-alpha), IL-6, and IL-8 were measured at diagnosis and 3 to 5 days after start of antibiotic treatment in 10 patients (mean age, 18.3 +/- 13.9 months) who had middle ear fluid culture positive for S. pneumoniae. The mean concentrations of all cytokines were elevated at diagnosis of AOM compared to levels in healthy controls, yet only IL-6 reached statistical significance (P = 0.05). IL-6 showed a statistically significant decrease in mean serum concentration at visit 2 (P = 0.03). IL-8 displayed a similar pattern to IL-6, but the difference between samples from day 1 and day 2 did not reach statistical significance. The cytokines IL-1 beta and TNF-alpha appear to be elevated in the serum of patients with S. pneumoniae AOM, but there was no significant change between mean serum levels obtained pre- and postinitiation of antibiotic treatment in the time frame studied. The results suggest a systemic inflammatory response as evidenced by increased IL-6. A significant decrease of IL-6 and improvement of clinical symptoms were observed. Determining cytokine levels, especially IL-6, in AOM could offer a powerful tool for objective assessment of response to treatment, minimizing unnecessary treatment of asymptomatic children who may still have some otoscopic findings suggestive of AOM at follow-up visits.

Topics: Biomarkers; Case-Control Studies; Child, Preschool; Clarithromycin; Follow-Up Studies; Humans; Infant; Inflammation; Interleukin-1; Interleukin-6; Interleukin-8; Interleukins; Otitis Media with Effusion; Pilot Projects; Pneumococcal Infections; Time Factors; Tumor Necrosis Factor-alpha

Otitis media with effusion (OME) may develop into a chronic course in some patients. In the majority of cases lipopolysaccharide (LPS) is detected in the middle ear fluid being capable of initiating and maintaining the inflammatory process associated with the disease. In addition, various extracellular signs of the inflammatory cascade are present, such as cytokines and ICAM-1 receptors. However, the underlying regulatory mechanisms situated intracellularily are sparsely recognized. NF-kappaB is a ubiquitously transcription factor complexed to its inhibitor within the cytoplasm. In response to stimuli, e.g. LPS or cytokines, NF-kappaB becomes activated and is translocated to the nucleus. At this level it induces transcription of various genes and subsequent expression of mRNA encoding for many immunoglobulins and cytokines, e.g. interleukin 8 (IL-8) which may be responsible for prolonging and maintaining the inflammatory process.. To evaluate if NF-kappaB is present in middle ear epithelium including to identify possible stimulating factors in vitro studies using rabbit middle ear epithelial cells (meec) were undertaken. Both normal cells and LPS exposed cells were studied. ELISA techniques were applied to detect NF-kappaB, ICAM-1 receptors and IL-8. All measurements were adjusted to the concentration of total cell protein (TP).. NF-kappaB was detected in the normal middle ear epithelium in concentrations between 16.8 and 28.6 ng/microg TP. In response to LPS the NF-kappaB content increased with 25-33%. This enhancement became more pronounced with longer duration of the LPS exposition. A relatively distinct expression of ICAM-1 preceded the NF-kappaB increase, after which the ICAM-1 measurements declined. IL-8 was hardly measurable in normal cells. The IL-8 concentrations were higher in LPS exposed cultures. The time related curve demonstrated a diphasic shape with an early and a late maximum.. The chronic inflammation seen in some OME patients may be due to LPS activating ICAM-1 receptors and NF-kappaB followed by release of IL-8. An autocrine pathway may be established through activation of TNF alpha and IL-1beta. Thus, elucidation of alternative pathways in the inflammatory cascade and elimination of LPS, alternatively inhibition of NF-kappaB and/or IL-8, should be an issue for future investigation.

Topics: Animals; Cells, Cultured; Chronic Disease; Ear, Middle; Enzyme-Linked Immunosorbent Assay; Epithelial Cells; Intercellular Adhesion Molecule-1; Interleukin-8; Lipopolysaccharides; NF-kappa B; Otitis Media with Effusion; Rabbits

Serous otitis media is usually responsive to medical treatment, whereas mucoid otitis media is not. The present study was undertaken to elucidate the compositional difference between serous and mucoid effusion and to investigate whether MUC5AC acts as a major mucin in the middle ear mucosa with mucoid otitis media.. This study involved a chemical analysis of middle ear effusion and immunostaining of the middle ear mucosa.. Middle ear effusion samples were collected from 27 patients with mucoid otitis media and 18 patients with serous otitis media. The levels of mucin, lysozyme, secretory immunoglobulin A, and interleukin-8 were measured by dot blotting or enzyme-linked immunosorbent assay. Periodic acid-Schiff and immunohistochemical staining with monoclonal anti-MUC5AC antibody were performed on the serial sections of middle ear mucosa with mucoid otitis media.. Mucoid effusions contained higher levels of mucin, lysozyme, secretory immunoglobulin A, and interleukin-8 than did serous effusions. Immunohistological study revealed that MUC5AC mucin was expressed in only a small portion of the goblet cells of middle ear mucosa with mucoid otitis media.. The study suggests that both serous secretions and mucin might make the middle ear effusion more viscous and that mucins other than MUC5AC might have a major role in the viscosity of middle ear effusion. Further study is necessary to identify the major mucins in the middle ear effusion of otitis media with effusion.

Topics: Child; Child, Preschool; Ear, Middle; Female; Humans; Immunoglobulin A, Secretory; Interleukin-8; Male; Mucin 5AC; Mucins; Mucous Membrane; Mucus; Muramidase; Otitis Media with Effusion; Viscosity

One of the main characteristics of otitis media with effusion (OME) is the differentiation of basal cells into goblet cells with subsequent proliferation in a modified respiratory epithelium leading to the formation of mucin-rich effusion in the middle ear cleft. In order to determine the effect of pro-inflammatory cytokines identified in OME, e.g. IL-1beta, tumour necrosis factor (TNF)-alpha, IL-6 and IL-8, on goblet cells, and to clarify the role of IL-8 in particular, we used the human goblet cell line HT29-MTX, which secretes two OME-related mucins: MUC5AC and MUC5B. IL-1beta and TNF-alpha stimulated the secretion of IL-8 in HT29-MTX goblet cells. Dose- (2-200 ng/ml) and time- (0-5 days) response studies of IL-8-induced mucin secretion were carried out. IL-8 upregulated the secretion of MUC5AC and MUC5B mucins in a concentration-dependent manner, with a maximum response at an IL-8 concentration of 20 ng/ml. IL-8 (20 ng/ml)-mediated mucin secretion persisted for up to 5 days, with a peak response 72 h after the addition of cytokine. These results suggest that: (i) goblet cells are target cells for the pro-inflammatory cytokines IL-1beta, TNF-alpha and IL-8 and can contribute to the pathogenesis of OME by increasing both the concentration of IL-8 and the secretion of mucin; and (ii) IL-8 stimulates prolonged mucin secretion from goblet cells and may be involved in the maintenance of the disease in the chronic stage.

Topics: Cell Differentiation; Cytokines; Ear, Middle; Goblet Cells; HT29 Cells; Humans; Inflammation Mediators; Interleukin-8; Mucins; Otitis Media with Effusion

To study the role of superoxide dismutase (SOD) and cytokines in the pathogenesis of secretory otitis media(SOM).. The content of SOD, interleukin-6(IL-6), interleukin-8(IL-8) and tumor necrosis factor alpha (TNF-alpha) in blood plasma and middle ear effusion (MEE) were measured in 64 (90 ears) patients respectively.. SOD, IL-6, IL-8 and TNF-alpha were found in 88.9%, 86.8%, 81.5% and 74.6% of MEI respectively. SOD, IL-6, IL-8 and TNF-alpha of blood plasma in SOM group were higher than those in normal control group (P < 0.05). The concentration of SOD, IL-6, IL-8 and TNF-alpha MEE was remarkably higher than that in blood plasma (P < 0.01). It revealed that the shorter the course, the higher the concentration of IL-6 or IL-8 in MEE (P < 0.01, or P < 0.05), and the longer the course, the higher the concentration of SOD or TNF-alpha in MEE (P < 0.01, P < 0.05). IL-6 and IL-8 concentration of MEE in serous fluid group was remarkably higher than those in the mucous fluid group (P < 0.01, P < 0.05), and the concentration of SOD and TNF-alpha of MEE in the mucous fluid group was higher than that in the serous fluid group (P < 0.01, P < 0.05). SOD Level of MEE was positively correlated with TNF-alpha (r = 0.587, P < 0.01).. SOD, IL-6, IL-8 and TNF-alpha might be important mediators in MEE secretion in SOM. IL-6 and IL-8 might participate in the defensive reaction of organism during the early stages of SOM. SOD and TNF-alpha might be closely related to the persistence of SOM, and might be mediators of mucin secretion in SOM.

Topics: Adolescent; Adult; Aged; Child; Cytokines; Female; Humans; Interleukin-6; Interleukin-8; Male; Middle Aged; Otitis Media with Effusion; Superoxide Dismutase; Tumor Necrosis Factor-alpha

Various inflammatory cells and cytokines have been identified in otitis media with effusion (OME). The presence of neutrophils has been linked to interleukin-8, but no chemotactic factor has as yet been identified for monocytes. The chemokine RANTES (Regulated upon Activation, Normal T Expressed and Secreted) attracts and activates primarily monocytes and may contribute to the pathogenesis of middle ear inflammation. We investigated the presence of RANTES by: 1) ELISA measurement in 114 middle ear effusions from children suffering from OME, 2) immunohistochemical localisation in experimental OME rabbit middle ear mucosa, and 3) expression in cultured rabbit middle ear epithelium in response to proinflammatory stimuli. RANTES was detectable in 94 (82%) of 114 effusions with a median concentration of 79.7 pg/mg total protein content. The concentration of RANTES was positively correlated with the endotoxin content. Immunohistochemically, RANTES was localized to the epithelial layer in experimental OME. In vitro, RANTES was expressed in middle ear epithelium in response to proinflammatory stimuli (TNF-alpha) in a dose-dependent manner. The expression of RANTES may explain the recruitment of monocytes in OME, possibly as a result of TNF-alpha-mediated endotoxin stimulation.

Topics: Animals; Cells, Cultured; Chemokine CCL5; Child; Cytokines; Disease Models, Animal; Ear, Middle; Endotoxins; Epithelium; Humans; Inflammation Mediators; Interleukin-8; Otitis Media with Effusion; Rabbits; Tumor Necrosis Factor-alpha

To study the role of cytokines and nitric oxide (NO) in the pathogenesis of secretory otitis media (SOM).. The content of NO, interleukin-8 (IL-8) and tumor necrosis factor alpha (TNF-alpha) in blood plasma and middle ear effusion (MEE) were measured in 58 (80 ears) patients with secretory otitis media and in 23 normal subjects, respectively.. NO, IL-8 and TNF-alpha were found in 100%, 82.2% and 71.4% of MEE respectively. NO, IL-8 or TNF-alpha of blood plasma in the SOM group was higher than those in the normal control group (P < 0.05). The concentration of NO, IL-8 or TNF-alpha in MEE was remarkably higher than those in blood plasma (P < 0.01). It revealed that the shorter the course, the higher the concentration of IL-8 in MEE (P < 0.01), and the longer the course, the higher the concentration of NO or TNF-alpha in MEE (P < 0.05). IL-8 concentration of MEE in serous fluid group was remarkably higher than that in the mucous fluid group (P < 0.01), and the concentration of NO or TNF-alpha of MEE in the mucous fluid group was higher than that in the serous fluid group (P < 0.05). NO level of MEE was positively correlated with TNF-alpha (r = 0.696 P < 0.01).. NO IL-8 and TNF-alpha might be important mediators in MEE secretion in SOM. IL-8 might participate in the defensive reaction of organism during the early stages of SOM, and play a role in serous effusion secretion. NO and TNF-alpha might be closely related to the persistence of SOM, and might be a mediator of mucin secretion in SOM.

Topics: Adolescent; Adult; Child; Child, Preschool; Exudates and Transudates; Female; Humans; Interleukin-8; Male; Middle Aged; Nitric Oxide; Otitis Media with Effusion; Tumor Necrosis Factor-alpha

adhesion of leukocytes to vascular endothelium and their invasion into local tissues are important steps in inflammation. L-selectin plays a crucial role in leukocyte rolling and adhesion on endothelial cell surface. Interleukin-8 (IL-8) is a potent chemotactic substance toward neutrophils and lymphocytes-T and mediate their migration to local inflammation. The levels of soluble L-selectin (sL-selectin) and IL-8 were measured in middle ear effusions (MEE) from children with otitis media (OM) to estimate their role in pathogenesis of inflammation in middle ear.. MEE were collected from 113 ears of 62 children during routine myringotomy. The entire specimen was diluted with phosphate-buffered saline (PBS) to 2 ml and centrifugated at 1500 rpm for 15 min to separate cellular components. Supernatants of MEE were stored frozen at -20 degrees C for sL-selectin and at -80 degrees C for IL-8 assessment. The concentration of sL-selectin and IL-8 were estimated with ELISA and compared with total protein concentration measured by Bradford assay.. MEEs from children with chronic mucous otitis media contained significantly higher mean concentrations of IL-8 559.4 pg/mg total protein (TP) (+/-535.6) in comparison with normal serum 17.79 pg/mg TP (+/-10.9), serous OM 40.3 pg/mg TP (+/-28.1) and purulent OM 104.4 pg/mg TP (+/-128.6). High concentration of IL-8 was found in MEE from the children with early recurrence of OMS. The levels of sL-selectin were higher in purulent effusions 77.2 ng/mg TP (+/-67.4) than those of chronic mucoid 27.6 ng/mg TP (+/-36.7) and serous OM 26.3 ng/mg TP (+/-27.1).. the results support a hypothesis that IL-8 can be responsible for prolongation of inflammatory process in middle ear, therefore long-term treatment and observation of children with the high levels of IL-8 in MEE may be necessary. Elevated level of sL-selectin in acute OM suggests the involvement of L-selectin in the acute phase of OM.

Topics: Adolescent; Child; Child, Preschool; Drainage; Female; Humans; Infant; Interleukin-8; L-Selectin; Male; Middle Ear Ventilation; Mucus; Osmolar Concentration; Otitis Media with Effusion; Proteins; Recurrence; Suppuration

To evaluate viral and cytokine signaling correlates of the persistent inflammation associated with chronic otitis media with effusion (OME).. Prospective study.. Reverse transcriptase-polymerase chain reaction targeting RNA viruses frequently associated with OME (respiratory syncytial virus and parainfluenza virus type 3, the proinflammatory cytokines interleukin 8 and interleukin 1beta, and RANTES [regulated upon activation, normal T cell expressed and secreted]) was performed on mucosal biopsy samples and on samples of the liquid and cellular compartments of inflammatory exudates obtained from 26 children (49 ears) with infected middle ears. Ribonucleic acid extracted from rapidly frozen samples was reverse transcribed by Moloney murine leukemia virus reverse transcriptase and amplified for 35 cycles using previously validated primers. Amplicons were evaluated by molecular size after agarose gel electrophoresis with ethidium bromide.. Most children had evidence of the presence of an RNA virus in at least one specimen. Respiratory syncytial virus was present in 40% and parainfluenza virus type 3 in 8% of effusions. Interleukin 8 messenger RNA was present in 21% of inflammatory exudates but never in cells from the mucosal biopsy samples.. Our data support a viral contribution to the cause of OME and suggest that the inflammatory cytokines observed derive more from cells in the inflammatory exudate than from those in the middle ear mucosa.

Topics: Biopsy; Chemokine CCL5; Child; Child, Preschool; Chronic Disease; Ear, Middle; Exudates and Transudates; Female; Humans; Interleukin-1; Interleukin-8; Male; Mucous Membrane; Otitis Media with Effusion; Parainfluenza Virus 3, Human; Polymerase Chain Reaction; Prospective Studies; Respiratory Syncytial Viruses; RNA, Messenger; RNA, Viral

We assayed 38 middle ear effusions from 23 children aged 4-13 years (mean 7) undergoing tympanostomy tube placements. All fluid was assayed for tumor necrosis factor (TNF) alpha, interleukin (IL) 1beta, IL-8, and IL-10. Cytokine concentrations were measured by means of an enzyme-linked immunosorbent assay. Detectable levels of IL-1beta, IL-8, and IL-10 were found in all of the effusions. TNF-alpha was detected in 18 of the middle ear effusions (47.4%). The mean concentration of TNF-alpha, IL-1beta , IL-8, and IL-10 was, respectively, 0.423 +/- 1.39, 30.58 +/- 68.7, 7001.9 +/- 6743, and 56 +/- 58.7 pg/ml. There was a strong, statistically significant correlation between the concentrations of TNF-alpha and IL-1beta (r = 0.87, P = 0.001) and between IL-1beta and IL-8 (r = 0.53, P = 0.001). There was no correlation between the concentrations of IL-10 and other cytokines examined or between tympanic membrane pathology and the concentrations of TNF-alpha, IL-1beta , IL-8, or IL-10. The presence of IL-10 in middle ear effusions may be one of the causes of a lack of clinical features of acute inflammation and may lead to a chronic inflammatory state.

Topics: Adolescent; Child; Child, Preschool; Cytokines; Enzyme-Linked Immunosorbent Assay; Female; Humans; Interleukin-1; Interleukin-10; Interleukin-8; Male; Middle Ear Ventilation; Otitis Media with Effusion; Tumor Necrosis Factor-alpha

The aim of this study was to clarify the site of primary pathology in otitis media with effusion. Effusions were collected from 64 children with bilateral effusions at the time of myringotomy. The rheological properties and biochemical compositions of effusions were measured for 23 pairs of effusions, and the levels of the inflammatory mediators TNF alpha, IL-1beta, and IL-8 were measured in 41 pairs using specific enzyme-linked immunosorbent assays (ELISAs). Measurements from paired ears were compared using analysis of variance (ANOVA) tests and significant differences were found for reduced specific viscosity, mucin content, protein content, and levels of IL-8. The results demonstrate that the two ears have different immunological processes or rates of processes which might explain the significantly different rheological properties of effusions. This suggests that each ear undergoes pathological changes independently and has implications for using the opposite ear as a control in clinical trials.

Topics: Analysis of Variance; Child; Child, Preschool; Enzyme-Linked Immunosorbent Assay; Exudates and Transudates; Humans; Hydrogen-Ion Concentration; Interleukin-1; Interleukin-8; Mucins; Otitis Media with Effusion; Proteins; Rheology; Tumor Necrosis Factor-alpha

The importance of interleukin (IL)-8 in the chemotactic activity of middle ear effusions (MEEs) was evaluated. There was a significantly higher IL-8 concentration in MEEs of children with acute otitis media (AOM) (n = 17; 136 ng/mL) than in children with otitis media with effusion (OME) (n = 28; 65 ng/mL). The IL-8 concentration in MEEs with bacteria (149 ng/mL) was significantly higher than in MEEs without bacteria (66 ng/mL). MEEs from children with AOM and OME had equally higher chemotactic activity than the diluent alone (23.3% and 24.8% vs. 9.2%). The chemotactic activity was not altered by the presence of bacteria nor did it correlate with IL-8 concentration. Fractionation of MEEs by gel chromatography demonstrated that the main chemotactic activity could clearly be separated from the IL-8 activity, thus excluding IL-8 as a main chemotactic component in MEEs.

Topics: Bacterial Infections; Chemotaxis; Child; Child, Preschool; Chromatography, Gel; Ear, Middle; Female; Humans; Infant; Interleukin-8; Male; Neutrophils; Otitis Media; Otitis Media with Effusion

Interleukin-8 (IL-8), a potent neutrophilic chemoattractant and inflammatory cytokine, is present in middle ear effusions (MEEs) of children with otitis media and is thought to be responsible for the accumulation of neutrophils in MEEs. We hypothesized that IL-8 concentration predicts the total number and proportion of neutrophils in MEEs. IL-8 concentration and total and differential cell counts were measured in MEEs of children undergoing tympanostomy tube placement for otitis media. IL-8 was present in 80 (98%) of 82 effusions. The mean +/- SEM value for IL-8 was 7342 +/- 847 pg/mL. The mean +/- SEM count and percentage of neutrophils were 1.34 x 10(6) +/- 3.44 x 10(5) and 70.6 +/- 3.1%, respectively. IL-8 concentrations correlated positively with the total number (r = +0.30; P = 0.02) and percentage of neutrophils (r = +0.32; P = 0.01) in the effusion. Additionally, purulent effusions had greater IL-8 concentrations (P = 0.003) and greater neutrophil count (P = 0.03) than mucoid or serous effusions. We conclude that IL-8 is consistently present in MEEs of children and IL-8 concentration predicts the total number and proportion of neutrophils. Furthermore, IL-8 concentration and the total number of neutrophils correlate positively with the type of effusion. These results support the hypothesis that IL-8 recruits neutrophils to the middle ear in MEEs.

Topics: Adolescent; Analysis of Variance; Cell Count; Chemotaxis, Leukocyte; Child; Child, Preschool; Enzyme-Linked Immunosorbent Assay; Female; Forecasting; Humans; Immunohistochemistry; Infant; Interleukin-8; Leukocyte Count; Lymphocyte Count; Macrophages; Male; Middle Ear Ventilation; Monocytes; Mucus; Neutrophils; Otitis Media with Effusion; Otitis Media, Suppurative

Interleukin-8 induces chemotaxis of neutrophils, basophils and T-lymphocytes, releases intracellular enzymes from neutrophils and histamine from basophils, and regulates the adhesion of neutrophils. In this study, using an enzyme-linked immunosorbent assay, we evaluated 33 middle ear effusions (MEEs) for levels of IL-8. The mean level of IL-8 in MEEs from children with OME was 616.7 +/- 211.0 pg/mgTP, while that of adults was 197.4 +/- 66.7 pg/mgTP. With respect to MEE types, the mean level of IL-8 in serous effusion was lower than that in two other types of MEEs (mucoserous and serous). These results suggest that inflammatory reaction in the middle ear cavity of children with OME is different from that of adults and that the pathogenesis of OME in children may differ from that in adults. Determination of IL-8 concentration in MEEs may help to illuminate the pathogenesis of OME.

Topics: Adult; Age Factors; Aged; Child; Child, Preschool; Enzyme-Linked Immunosorbent Assay; Exudates and Transudates; Female; Humans; Interleukin-8; Male; Middle Aged; Otitis Media with Effusion; Proteins; Reagent Kits, Diagnostic; Spectrophotometry

Based on recent studies in the authors' laboratory on the correlation of cytokines and inflammation in otitis media (OM), the authors hypothesized that in chronic otitis media with effusion (COME) interleukin-8 (IL-8) is responsible for 1. the accumulation of leukocytes in the middle ear cleft and 2. in situ leukocyte activation with subsequent tissue damage. Additionally, the authors hypothesized that IL-8 expression is at least in part under the control of interleukin-1 (IL-1) and tumor necrosis factor (TNF). To begin to test this hypothesis, middle ear effusions (MEE) obtained from children ages 2 to 90 months (mean age, 29 months) undergoing tympanostomy tube placement for the presence of these inflammatory cytokines were analyzed. For these studies, IL-8, interleukin-1 beta (IL-1 beta), tumor necrosis factor-alpha (TNF-alpha), and tumor necrosis factor-beta (TNF-beta) were measured in MEE by radioimmunoassay (RIA) or enzyme-linked immunoassay (ELISA). IL-8, IL-1 beta, TNF-alpha, and TNF-beta were present in 92%, 67%, 77%, and 0% of effusions, respectively. The mean (+/- SEM) values for IL-8, IL-1 beta, and TNF-alpha were 4805 (+/- 913) pg/mg, 4076 (+/- 1510) pg/mg, and 163 (+/- 90) pg/mg. Further analysis indicated that levels of IL-8 correlated with IL-1 beta (R2 = .500, P = .000) and TNF-alpha (R2 = .387, P = .023). Thus the authors' studies clearly demonstrate that IL-8 is consistently present in the MEE of children with COME and is strongly correlated with levels of IL-1 beta and TNF-alpha, both known inducers of IL-8 production. These results support the authors' hypothesis that IL-1 beta, TNF-alpha, and IL-8 are intimately involved in the inflammatory cascade in the middle ear and suggest regulation of these cytokines as possible sites of future therapeutic intervention in otitis media with effusion (OME).

Topics: Age Factors; Child; Child, Preschool; Chronic Disease; Cytokines; Female; Gene Expression; Humans; Infant; Interleukin-1; Interleukin-8; Male; Middle Ear Ventilation; Otitis Media with Effusion; Proteins; Recurrence; Tumor Necrosis Factor-alpha

Interleukin-8 (IL-8), a monocyte- and macrophage-derived cytokine, displays potent chemotactic-activating properties toward neutrophils, and thus may contribute to the pathogenesis of otitis media with effusion (OME). The objective of this investigation was to demonstrate the expression of the IL-8 gene in middle ear effusion (MEEs) of children and adults with OME. Ribonucleic acids (RNAs) were extracted from MEEs from 16 ears of 13 pediatric patients and 12 ears of 12 adult patients with OME. The RNAs were reverse-transcribed and amplified by the polymerase chain reaction. Interleukin-8 transcripts were detected in 75% of both pediatric (12/16) and adult MEEs (9/12). The levels of expression of IL-8 and beta-actin messenger RNAs were quantitated. No significant difference was observed in IL-8/beta-actin ratios between pediatric MEEs and adult MEEs. These data suggest that IL-8 may contribute to neutrophil involvement in both pediatric and adult OME.

Topics: Actinin; Adolescent; Adult; Aged; Child; Child, Preschool; Cytokines; Ear, Middle; Gene Expression; Humans; Interleukin-8; Middle Aged; Neutrophils; Oligonucleotides; Otitis Media with Effusion; RNA, Messenger; Transcription, Genetic

Otitis media with effusion (OME) is a common middle ear inflammatory disease in the pediatric population. This article determines concentrations of three functionally and metabolically distinct inflammatory mediators in middle ear effusions (MEE) and corresponding plasma of children with OME. One hundred two patients (mean age, 4.9 years) with persistent OME were studied. Middle ear effusions were collected from all subjects and plasma from a subset at the time of tympanostomy tube insertion. Histamine was assayed radioisotopically, 13,14-dihydro-15-keto-prostaglandin F2 alpha (stable PGF2 alpha metabolite) by radioimmunoassay, and neutrophil chemotactic factor of anaphylaxis by modified Boyden chamber. Mean MEE levels of the mediators (39 +/- 13 ng/mL, 462 +/- 179 pg/mL, and 264% +/- 57% positive control, respectively) were markedly higher than those of corresponding plasma (0.5 +/- 0.1 ng/mL, 285 +/- 127 pg/mL, and 47% +/- 5% positive control, respectively). The mean histamine content of mucoid effusions (43.2 +/- 56.9 ng/mL) was significantly higher than that of purulent (22.5 +/- 10.5 ng/mL) and serous (17.9 +/- 16.8 ng/mL) effusions. Higher histamine levels were observed in effusions positive for Haemophilus influenzae when compared with those with other pathogenic isolates. The high concentrations of these mediators in MEE and their potential for inducing or sustaining the inflammatory process supports a role in the pathogenesis of OME.

Topics: Adolescent; Adult; Chemotactic Factors; Child; Child, Preschool; Chronic Disease; Dinoprost; Haemophilus influenzae; Histamine; Humans; Infant; Inflammation; Interleukin-8; Otitis Media with Effusion; Prostaglandins F