interleukin-8 and Rhinitis--Allergic--Seasonal

The allergic process is believed to consist of two phases: early and late. The early phase reaction is mainly induced by histamine released from mast cells. Histamine binding specific cell receptors produces clinical allergic symptoms. This mediator also activates neutrophils and eosinophils as well as being a chemoattractant for these cells. Histamine increases IL-8 level and evokes leukocyte rolling on endothelial cells. Thus histamine participates in both early and late-phase allergic responses.

Topics: Binding Sites; Chemoreceptor Cells; Eosinophils; Histamine; Humans; Inflammation Mediators; Interleukin-8; Leukotriene B4; Mast Cells; Neutrophils; Receptors, Histamine; Rhinitis, Allergic, Seasonal

Interleukin (IL)-33 is a novel member of the IL-1 cytokine family and a ligand for the orphan IL-1 family receptor ST2. The IL-33 induces T helper 2-type inflammatory responses and is considered to play a crucial rule in allergic inflammations, such as asthma and atopic dermatitis. However, the role of IL-33 and its receptor ST2 in allergic rhinitis remains unknown.. We investigated expression of IL-33 and ST2 in the nasal epithelium of patients with allergic rhinitis and the mechanisms of the production of cytokines/chemokines induced by treatment with IL-33 using normal human nasal epithelial cells (HNECs) in vitro.. Expression of IL-33 and ST2 in normal and allergic rhinitis nasal mucosa was evaluated by reverse transcription- and real-time polymerase chain reactions and immunohistochemical methods. The IL-33 in serum, and IL-8 and GM-CSF were measured by ELISA. For in vitro experiments, HNECs in primary culture were used.. The IL-33 levels in the sera of patients with allergic rhinitis were significantly higher than that in normal controls. Expression of IL-33 and ST2 was significantly elevated in the epithelium from patients with allergic rhinitis. The IL-33 mRNA in HNECs in vitro was significantly induced by treatment with IFN-γ and the toll-like receptor 9 ligand ODN2006. The IL-33-induced production of IL-8 and GM-CSF from HNECs in vitro was significantly suppressed by corticosteroid treatment and distinct signal transduction inhibitors of ERK, p38 MAPK, JNK, NF-κB and epidermal growth factor receptor.. The IL-33 and its receptor ST2 play important roles in allergic rhinitis. The IL-33-mediated inflammatory responses via ST2 are regulated by distinct signalling pathways in HNECs and the IL-33/ST2 pathway may provide new therapeutic targets for allergic rhinitis.

Topics: Adolescent; Adult; Aged; Antiviral Agents; Cells, Cultured; ErbB Receptors; Female; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Interferon-gamma; Interleukin-1 Receptor-Like 1 Protein; Interleukin-33; Interleukin-8; Interleukins; Male; MAP Kinase Kinase 4; MAP Kinase Signaling System; Middle Aged; Nasal Mucosa; NF-kappa B; p38 Mitogen-Activated Protein Kinases; Real-Time Polymerase Chain Reaction; Receptors, Cell Surface; Reverse Transcriptase Polymerase Chain Reaction; Rhinitis, Allergic, Seasonal; RNA, Messenger; Toll-Like Receptor 9; Young Adult

Allergic rhinitis symptoms of itching, sneezing, rhinorrhea, and nasal obstruction significantly decrease patients' quality of life. Compared with histamine and leukotriene receptor antagonists, the petasol butenoate complex Ze 339 displays pharmacologically distinct properties. In vitro it inhibits the biosynthesis of leukotrienes and mediator release from activated eosinophils.. This study aimed to assess the efficacy and mode of action of Ze 339, desloratadine, and placebo on allergic rhinitis symptoms, nasal airflow, and local mediator levels after unilateral nasal allergen provocation.. In this double-blind, randomized, crossover study 18 subjects with allergic rhinitis to grass pollen received Ze 339, desloratadine, and placebo for 5 days before nasal allergen challenge with grass pollen extract. Rhinomanometry, symptom assessment, and local inflammatory mediator measurement were performed during the 24 hours after allergen challenge.. With Ze 339, the patient's time to recovery (5.4 ± 1.6 hours) from nasal obstruction after allergen challenge (time for return to 90% of baseline value ± SEM) was significantly shorter than with placebo (9.1 ± 2.3 hours, P = .035) and desloratadine (10.7 ± 2.5 hours, P = .022). Likewise, Ze 339's standardized symptom assessment for nasal obstruction (3.2 ± 1.3 hours) showed significantly faster relief (time for return to baseline value ± SEM compared with placebo, 8.3 ± 2.4 hours; P = .027) and desloratadine (4.5 ± 1.2 hours, P = .030). One interesting finding was that Ze 339 significantly reduced IL-8 and leukotriene B(4) levels in nasal secretions before challenge.. When compared with desloratadine and placebo, Ze 339 shows better efficacy in relieving nasal obstruction symptoms and inhibiting critical components of the chemokine network and as such represents a novel symptomatic and possible prophylactic treatment for allergic rhinitis.

Topics: Adult; Allergens; Anti-Allergic Agents; Bronchial Provocation Tests; Chemokines; Cross-Over Studies; Cytokines; Double-Blind Method; Female; Humans; Interleukin-8; Leukotriene B4; Loratadine; Male; Middle Aged; Nasal Obstruction; Plant Extracts; Pollen; Rhinitis, Allergic, Seasonal; Young Adult

We have previously demonstrated the presence of toll-like receptor 9 in the nasal mucosa of both healthy and allergic individuals. CpG motifs, found in bacterial and viral DNA, elicit strong immunostimulatory effects via this receptor. CpG is known to skew the immune system towards a T helper 1 (Th1) profile, thereby suppressing Th2-driven allergic responses. This study was designed to examine the effects of CpG administration in the human nose.. Twenty subjects, of whom 10 suffered from seasonal allergic rhinitis (AR), were challenged intranasally with CpG outside pollen season. Symptom scores, nasal airway resistance (NAR), and nasal and pulmonary nitric oxide (NO) levels were assayed prior to challenge and 30 min, 6, 24 and 48 h post challenge. The presence of leukocytes and various cytokines were analyzed in nasal lavage (NAL) fluids before and after CpG exposure.. Increased NAR, nasal NO production and secretion of interleukin (IL)-1beta, IL-6, and IL-8 were seen after CpG exposure. Further analysis revealed that this inflammatory response was more marked in healthy subjects than among patients with AR, although a higher basal inflammatory response was recorded in the allergic group. In vitro experiments suggest that the effects induced by CpG are mediated by epithelial cells and neutrophils.. Nasal administration of CpG induces a local airway inflammation, more distinct among healthy than allergic individuals. The reduced responsiveness to CpG in allergic patients might be related to the ongoing minimal persistent inflammation. Results from cytokine analyses reflect the ability of CpG to induce a pro-inflammatory Th1-like immune response.

Topics: Adjuvants, Immunologic; Administration, Intranasal; Adult; Allergens; Cell Line; Cytokines; Female; Humans; Interleukin-1beta; Interleukin-6; Interleukin-8; Male; Middle Aged; Nasal Lavage Fluid; Nasal Mucosa; Neutrophils; Nitric Oxide; Oligodeoxyribonucleotides; Rhinitis, Allergic, Seasonal; Skin Tests; Toll-Like Receptor 9

TNFalpha may contribute to the pathophysiology of airway inflammation. For example, we have recently shown that nasal administration of TNFalpha produces late phase co-appearance of granulocyte and plasma exudation markers on the mucosal surface. The objective of the present study was to examine indices of granulocyte presence and activity in response to intranasal TNFalpha challenge.. Healthy subjects and patients with allergic rhinitis (examined out of season) were subjected to nasal challenge with TNFalpha (10 microg) in a sham-controlled and crossover design. Nasal lavages were carried out prior to and 24 hours post challenge. Nasal biopsies were obtained post challenge. Nasal lavage fluid levels of myeloperoxidase (MPO) and eosinophil cationic protein (ECP) were analyzed as indices of neutrophil and eosinophil activity. Moreover, IL-8 and alpha2-macroglobulin were analyzed as markers of pro-inflammatory cytokine production and plasma exudation. Nasal biopsy numbers of neutrophils and eosinophils were monitored.. Nasal lavage fluid levels of MPO recorded 24 hours post TNFalpha challenge were increased in healthy subjects (p = 0.0081) and in patients with allergic rhinitis (p = 0.0081) (c.f. sham challenge). Similarly, alpha2-macroglobulin was increased in healthy subjects (p = 0.014) and in patients with allergic rhinitis (p = 0.0034). Lavage fluid levels of ECP and IL-8 were not affected by TNFalpha challenge. TNFalpha increased the numbers of subepithelial neutrophils (p = 0.0021), but not the numbers of eosinophils.. TNFalpha produces a nasal inflammatory response in humans that is characterised by late phase (i.e., 24 hours post challenge) neutrophil activity and plasma exudation.

Topics: Administration, Intranasal; Adult; alpha-Macroglobulins; Biopsy; Cross-Over Studies; Eosinophil Cationic Protein; Eosinophils; Female; Granulocytes; Humans; Interleukin-8; Male; Middle Aged; Nasal Lavage Fluid; Nasal Mucosa; Neutrophils; Peroxidase; Pneumonia; Rhinitis, Allergic, Seasonal; Single-Blind Method; Tumor Necrosis Factor-alpha

beta-Tryptase is a multifunctional mast cell serine protease released during mast cell degranulation and tryptase/trypsin inhibitors are a novel potential therapeutic approach for allergic inflammatory diseases.. This study was performed to assess the effects of RWJ-58643 on nasal symptoms, eosinophil influx, and cytokine and chemokine release following nasal allergen challenge (NAC).. Male patients with grass pollen allergic rhinitis (n=16) out of season received single doses of RWJ-58643 (100, 300, 600 microg) or matched placebo given 30 min before NAC in a double-blind, randomized crossover design. A single dose of 200 microg budesonide was studied in an open-label extension phase. NAC was performed with Timothy grass pollen (ALK) via a nasal device, and nasal lavage was performed at times 0 (pre-drug, pre-allergen), 0.5 (30 min post-drug, pre-NAC) 1.5, 2.5, 4.5, 6.5, 8.5, and 24 h after drug administration. Nasal lavage mediators were analysed using a sensitive multiplexed bead immunoassay system.. Low-dose RWJ-58643 (100 microg) and budesonide (200 microg) significantly reduced symptoms, eosinophils and levels of IL-5 following NAC. However, higher doses of RWJ-58643 (300 and 600 microg) caused a late eosinophilia and preceding increases in IL-5 compared with placebo.. This study suggests that combined beta-tryptase and trypsin inhibition has therapeutic potential in allergic inflammation, however, this property is dose responsive and higher doses are ineffective and may cause eosinophilia.

Topics: Administration, Intranasal; Adult; Allergens; Benzothiazoles; Budesonide; Chemokine CCL11; Chemokine CCL2; Chemokines, CC; Chemotactic Factors, Eosinophil; Cross-Over Studies; Double-Blind Method; Eosinophils; Female; Humans; Inflammation Mediators; Interleukin-5; Interleukin-8; Leukocyte Count; Male; Mast Cells; Middle Aged; Pyrrolidines; Rhinitis, Allergic, Seasonal; Serine Endopeptidases; Thiazoles; Trypsin Inhibitors; Tryptases; Tumor Necrosis Factor-alpha

Airway hyperresponsiveness and inflammation can be noninvasively studied by bronchial provocation using direct (histamine) or indirect (adenosine 5'-monophosphate [AMP]) stimuli and induced sputum.. To report on the immediate effects of histamine and AMP challenge on induced sputum neutrophil counts and related mediator levels.. We performed a single-masked, randomized, placebo-controlled, 3-way, crossover, methodological study in 14 atopic patients (median age, 25 years; 8 males; mean +/- SD forced expiratory volume in 1 second, 99% +/- 5%) without anti-inflammatory medication use. At baseline, sputum induction was performed. Bronchial challenges with AMP, histamine, or placebo were performed 48 hours later. Thirty minutes after challenge, sputum induction was performed again. Challenge periods in each patient were separated by more than 2 weeks. Sputum cells and the mediators leukotriene B4, interleukin 8, myeloperoxidase, and albumin were quantified.. Comparing median challenge-induced relative changes in cells and mediators, neither histamine nor AMP challenge altered the induced sputum neutrophil counts (histamine, 2.7%; AMP, 2.95%; placebo, -2%; P > .07 for all), interleukin 8 levels (histamine, 2.4 ng/mL; AMP, -3.8 ng/mL; placebo, -0.2 ng/mL; P > .06), leukotriene B4 levels (histamine, -4.8 pg/mL; AMP, 3 pg/mL; placebo, 6 pg/mL; P > .08), or myeloperoxidase levels (histamine, 0.16 microg/mL; AMP, 0 microg/mL; placebo, -0.03 microg/mL; P > .07). Sputum albumin levels were increased after histamine challenge compared with AMP and placebo challenge (P < .01 for both).. Histamine and AMP provocation have no major effects on induced neutrophil counts and related mediator levels in atopic patients, whereas histamine challenge induces plasma leakage. Potential interactions of noninvasive methods to evaluate airway reactivity and inflammation should be carefully considered.

Topics: Adenosine Monophosphate; Adult; Asthma; Bronchial Provocation Tests; Cell Count; Cross-Over Studies; Female; Histamine; Humans; Interleukin-8; Leukotriene B4; Male; Neutrophils; Peroxidase; Rhinitis, Allergic, Seasonal; Sputum

Extract of Perilla frutescens enriched for rosmarinic acid, a polyphenolic phytochemical, suppresses allergic immunoglobulin responses and inflammation caused by polymorphonuclear leukocytes (PMNL) in mice. However, few placebo-controlled clinical trials have examined the efficacy and safety of polyphenolic phytochemicals for treatment of allergic inflammatory diseases in humans. The present study determined whether oral supplementation with rosmarinic acid is an effective intervention for patients with seasonal allergic rhinoconjunctivitis (SAR). In this 21-day, randomized, double-blind, age-matched, placebo-controlled parallel group study, patients with mild SAR were treated daily with extract of Perilla frutescens enriched for rosmarinic acid (200 mg [n=10] or 50 mg [n=9]) or placebo (n=10). Patients recorded symptoms daily in a diary. Profiles of infiltrating cells and concentrations of eotaxin, IL-1beta, IL-8, and histamine were measured in nasal lavage fluid. Serum IgE concentrations and routine blood tests were also examined. As compared with placebo supplementation, supplementation with extract of Perilla frutescens enriched for rosmarinic acid resulted in a significant increase in responder rates for itchy nose, watery eyes, itchy eyes, and total symptoms (P<0.05). Active treatment significantly decreased the numbers of neutrophils and eosinophils in nasal lavage fluid (P<0.05 vs. placebo). Patients reported no adverse events, and no significant abnormalities were detected in routine blood tests. In conclusion, extract of Perilla frutescens enriched for rosmarinic acid can be an effective intervention for mild SAR at least partly through inhibition of PMNL infiltration into the nostrils. Use of this alternative treatment for SAR might reduce treatment costs for allergic diseases.

Topics: Adult; Anti-Inflammatory Agents, Non-Steroidal; Chemokine CCL11; Chemokines, CC; Cinnamates; Conjunctivitis, Allergic; Depsides; Double-Blind Method; Eosinophils; Female; Histamine; Humans; Hypersensitivity, Immediate; Immunoglobulin E; Interleukin-1; Interleukin-8; Male; Middle Aged; Nasal Lavage Fluid; Neutrophils; Perilla frutescens; Phytotherapy; Plant Preparations; Rhinitis, Allergic, Seasonal; Rosmarinic Acid; Treatment Outcome

Allergic rhinitis is characterized by an IgE-dependent inflammation. Nasal obstruction is related to allergic inflammation. Some antihistamines have been demonstrated to be capable of improving this nasal symptom.. The aim of this pilot study was to evaluate nasal symptoms, nasal airflow, inflammatory cells, and cytokine pattern in patients with seasonal allergic rhinitis (SAR), before and after treatment with levocetirizine, desloratadine, or placebo.. Thirty patients with SAR were evaluated, 27 males and three females (mean age 26.9+/-5.4 years). All of them received levocetirizine (5 mg/day), desloratadine (5 mg/day), or placebo for 2 weeks. The study was double-blind, parallel-group, placebo-controlled, and randomized. Total symptom score (TSS) (including: rhinorrhea, nasal itching, sneezing, and nasal obstruction) was assessed before and after treatment. Rhinomanometry, nasal lavage, and nasal scraping were performed in all subjects before and after treatment. Inflammatory cells were counted by conventional staining; IL-4 and IL-8 were measured by immunoassay on fluids recovered from nasal lavage.. Levocetirizine treatment induced significant symptom relief (P=0.0009) and improved nasal airflow (P=0.038). Desloratadine also relieved TSS (P=0.01), but did not affect nasal airflow. Levocetirizine significantly reduced eosinophils (P=0.029), neutrophils (P=0.005), IL-4 (P=0.041), and IL-8 (P=0.02), whereas desloratadine diminished IL-4 only (P=0.044). Placebo treatment did not significantly affect any evaluated parameters.. This pilot study demonstrates the effectiveness of levocetirizine in: (i) relieving nasal symptoms, (ii) improving nasal airflow, (iii) reducing leucocyte infiltration, and (iv) diminishing cytokine levels. These findings are the first evidence of the effectiveness of levocetirizine in SAR.

Topics: Adult; Cetirizine; Double-Blind Method; Eosinophils; Female; Histamine H1 Antagonists; Humans; Interleukin-4; Interleukin-8; Leukocyte Count; Loratadine; Male; Nasal Lavage Fluid; Nasal Obstruction; Pilot Projects; Piperazines; Rhinitis, Allergic, Seasonal; Statistics, Nonparametric

Binding of allergens to IgE on mast cells and basophils causes release of inflammatory mediators in nasal secretions.. The combined effect of specific immunotherapy (SIT) and omalizumab, a humanized monoclonal anti-IgE antibody, on release of eosinophilic cationic protein (ECP), tryptase, IL-6, and IL-8 in nasal secretion was evaluated.. Two hundred and twenty five children (aged 6-17 years) with a history of seasonal allergic rhinoconjunctivitis induced by birch and grass pollen were randomized into four groups: either birch- or grass-pollen SIT in combination with either anti-IgE or placebo. Complete sets of nasal secretion samples before treatment Visit 1 (V1), during birch- (V2) and grass (V3)-pollen season and after the pollen season (V4) were collected from 53 patients.. A significant reduction in tryptase only was seen in the anti-IgE-treated group at V2 (P<0.05) and V4 (P<0.05) compared with the placebo group. During the pollen season, patients with placebo showed an increase of ECP compared with baseline (V2: +30.3 microg/L; V3: +134.2 microg/L, P< 0.005; V4: +79.0 microg/L, P< 0.05), and stable levels of tryptase, IL-6 and IL-8. Treatment with anti-IgE resulted in stable ECP values and a significant decrease of tryptase compared with V1 (baseline): V2: -80.0 microg/L (P< 0.05); V3: -56.3 microg/L, which persisted after the pollen season with V4: -71.6 microg/L (P< 0.05). After the pollen season, a decrease of IL-6 was observed in both groups (V4 placebo group: -37.5 ng/L; V4 anti-IgE group: -42.9 ng/L, P< 0.01).. The combination of SIT and anti-IgE is associated with prevention of nasal ECP increase and decreased tryptase levels in nasal secretions.

Topics: Adolescent; Antibodies, Anti-Idiotypic; Antibodies, Monoclonal; Antibodies, Monoclonal, Humanized; Betula; Blood Proteins; Body Fluids; Child; Desensitization, Immunologic; Double-Blind Method; Eosinophil Granule Proteins; Female; Humans; Immunoglobulin E; Interleukin-6; Interleukin-8; Male; Nasal Mucosa; Omalizumab; Poaceae; Pollen; Rhinitis, Allergic, Seasonal; Ribonucleases; Serine Endopeptidases; Tryptases

Cetirizine has been demonstrated able of reducing nasal inflammatory infiltration in children with allergic rhinitis and cytokine production in in vitro studies. The aim of this double-blind, placebo-controlled, and randomized study was to evaluate cytokine pattern and inflammatory cells in children with perennial allergic rhinitis, before and after treatment with cetirizine or placebo. Twenty children with perennial allergic rhinitis were evaluated, 13 males and 7 females (mean age 13.4 years). Inflammatory cells and cytokines were evaluated by scraping and nasal lavage, before and after 2-weeks administration of cetirizine or placebo. IL4 and IL8 were measured by immunoassay and inflammatory cells were counted by conventional staining. Cetirizine treatment induced a significant decrease of IL4 (p<0.01) and IL8 levels (p=0.01). A significant reduction of the inflammatory cells was detected in actively-treated children, both concerning neutrophils and eosinophils (p<0.01). Moreover, cetirizine significantly reduced nasal obstruction score (p=0.007). This study shows the cetirizine effectiveness in exerting anti-inflammatory activity by modulating cytokine pattern and by reducing inflammatory infiltration in children with perennial allergic rhinitis.

Topics: Adolescent; Animals; Cetirizine; Child; Double-Blind Method; Enzyme-Linked Immunosorbent Assay; Eosinophils; Female; Histamine H1 Antagonists, Non-Sedating; Humans; Interleukin-4; Interleukin-8; Male; Nasal Lavage Fluid; Nasal Mucosa; Nasal Obstruction; Neutrophils; Pollen; Prospective Studies; Pyroglyphidae; Rhinitis, Allergic, Perennial; Rhinitis, Allergic, Seasonal; Treatment Outcome

Interleukin-8 (IL-8) is a chemokine that causes chemotaxis of neutrophils, eosinophils and lymphocytes in vitro; however, its role as a chemoattractant in allergic inflammation is unclear. The objective of this study was to investigate the effect of nasal instillation of IL-8 on the influx of inflammatory cells.. Twelve patients suffering from seasonal allergic rhinitis hypersensitive to grass pollens, with average age 30.1 +/- 2.67 years were challenged both with diluent for IL-8 and IL-8 on a subsequent day, in two phases: before the pollen season (unprimed mucosa) and during the season (primed mucosa). The number of neutrophils, eosinophils and myeloperoxydase (MPO) levels in the nasal fluid collected after IL-8 or placebo challenge were determined.. Challenge with IL-8 of primed nasal mucosa induced a significant influx of neutrophils (29 x 10(4) cells/ml at 0.5 h, 251 x 10(4) at 2 h and 334 x 10(4) at 3 h). Number of eosinophils in comparison with diluent challenge was not significant. There was no difference in MPO levels in the nasal lavage between IL-8 and diluent challenge of unprimed and primed mucosa. We did not find the relationship between MPO levels and the neutrophils number in the lavage (rank Spearman factor, RS = 0.258, P = 0.42).. We have demonstrated that IL-8 causes influx of neutrophils but not eosinophils into nasal mucosa in vivo. MPO level seems to be of little value as a marker of neutrophil influx into nasal mucosa.

Topics: Administration, Intranasal; Adult; Cell Count; Chemotaxis, Leukocyte; Eosinophils; Female; Humans; Interleukin-8; Male; Middle Aged; Nasal Lavage Fluid; Nasal Mucosa; Neutrophils; Peroxidase; Recombinant Proteins; Rhinitis, Allergic, Seasonal

Recent studies have suggested that exposure to air pollutants may enhance the airway responsiveness of susceptible individuals to inhaled allergen.. To investigate the effect of exposure to nitrogen dioxide (NO2) on nasal airways resistance (NAR) and inflammatory mediators in nasal lavage fluid, eight subjects with a history of seasonal allergic rhinitis, who were tested out of season, were exposed in a randomized single-blind, crossover study to either air or 400 ppb NO2 for 6 hours. The changes in NAR and eosinophil cationic protein (ECP), mast cell tryptase (MCT), neutrophil myeloperoxidase (MPO), and interleukin-8 (IL-8) in nasal lavage fluid before and after exposure were evaluated. Another group of eight subjects with a history of seasonal allergic rhinitis were also randomized to exposure to air or 400 ppb NO2 for 6 hours and then challenged with allergen, before evaluation for changes in NAR and changes in ECP, MCT, MPO, and IL-8 in nasal lavage fluid.. Exposure to air or NO2 did not alter either NAR or the levels of ECP, MCT, MPO, or IL-8 in nasal lavage fluid. Allergen challenge after exposure to both air and NO2 significantly (p < 0.05) increased levels of MCT, but not MPO and IL-8 in the nasal lavage fluid. In addition, allergen challenge after exposure to NO2 but not air, significantly increased levels of only ECP in nasal lavage fluid (p < 0.05).. These results suggest that acute exposure to NO2 at concentrations found at the curbside in heavy traffic during episodes of pollution, may "prime" eosinophils for subsequent activation by allergen in individuals with a history of seasonal allergic rhinitis.

Topics: Adolescent; Adult; Airway Resistance; Allergens; Blood Proteins; Chymases; Cross-Over Studies; Eosinophil Granule Proteins; Female; Humans; Inflammation Mediators; Interleukin-8; Male; Mast Cells; Middle Aged; Nasal Lavage Fluid; Nasal Mucosa; Nasal Provocation Tests; Nebulizers and Vaporizers; Nitrogen Dioxide; Peroxidase; Pollen; Rhinitis, Allergic, Seasonal; Ribonucleases; Serine Endopeptidases; Single-Blind Method; Tryptases

Pollen exposure induces local and systemic allergic immune responses in sensitized individuals, but nonsensitized individuals also are exposed to pollen. The kinetics of symptom expression under natural pollen exposure have never been systematically studied, especially in subjects without allergy.. We monitored the humoral immune response under natural pollen exposure to potentially uncover nasal biomarkers for in-season symptom severity and identify protective factors.. We compared humoral immune response kinetics in a panel study of subjects with seasonal allergic rhinitis (SAR) and subjects without allergy and tested for cross-sectional and interseasonal differences in levels of serum and nasal, total, and Betula verrucosa 1-specific immunoglobulin isotypes; immunoglobulin free light chains; cytokines; and chemokines. Nonsupervised principal component analysis was performed for all nasal immune variables, and single immune variables were correlated with in-season symptom severity by Spearman test.. Symptoms followed airborne pollen concentrations in subjects with SAR, with a time lag between 0 and 13 days depending on the pollen type. Of the 7 subjects with nonallergy, 4 also exhibited in-season symptoms whereas 3 did not. Cumulative symptoms in those without allergy were lower than in those with SAR but followed the pollen exposure with similar kinetics. Nasal eotaxin-2, CCL22/MDC, and monocyte chemoattactant protein-1 (MCP-1) levels were higher in subjects with SAR, whereas IL-8 levels were higher in subjects without allergy. Principal component analysis and Spearman correlations identified nasal levels of IL-8, IL-33, and Betula verrucosa 1-specific IgG. Nasal pollen-specific IgA and IgG isotypes are potentially protective within the humoral compartment. Nasal levels of IL-8, IL-33, sIgG

Topics: Adult; Allergens; Antigens, Plant; Biomarkers; Female; Humans; Immunoglobulin A; Immunoglobulin E; Immunoglobulin G; Interleukin-33; Interleukin-8; Male; Middle Aged; Nasal Mucosa; Pollen; Rhinitis, Allergic, Seasonal; Seasons; Young Adult

Topics: Ambrosia; Animals; Blood-Air Barrier; Humans; Interleukin-8; Neutrophil Infiltration; Neutrophils; NF-kappa B; Peptide Hydrolases; Pollen; Receptor, PAR-2; Receptors, G-Protein-Coupled; Receptors, Interleukin-8B; Rhinitis, Allergic, Seasonal; Toll-Like Receptor 4

Besides allergens, pollen release bioactive, low molecular weight compounds that modulate and stimulate allergic reactions. Clinical relevance of these substances has not been investigated to date.. To elucidate the effect of a non-allergenic, low molecular weight factors from aqueous birch pollen extracts (Bet-APE < 3 kDa) on the human allergic immune response in vivo.. Birch and grass pollen allergic individuals underwent skin prick testing with allergen alone, allergen plus Bet-APE < 3 kDa, or allergen plus pre-identified candidate substances from low molecular pollen fraction. Nasal allergen challenges were performed in non-atopic and pollen allergic individuals using a 3 day repeated threshold challenge battery. Subjects were either exposed to allergen alone or to allergen plus Bet-APE< 3 kDa. Local cytokine levels, nasal secretion weights, nasal congestion and symptom scores were determined.. Skin prick test reactions to pollen elicited larger weals when allergens were tested together with the low molecular weight compounds from pollen. Similar results were obtained with candidate pollen-associated lipid mediators. In nasal lining fluids of allergic patients challenged with allergen plus Bet-APE < 3 kDa, IL-8 and IgE was significantly increased as compared to allergen-only challenged patients. These patients also produced increased amounts of total nasal secretion and reported more severe rhinorrhea than the allergen-only challenged group.. Low molecular compounds from pollen enhance the allergen specific immune response in the skin and nose. They are therefore of potential clinical relevance in allergic patients.

Topics: Allergens; Basophils; Betula; Cell Degranulation; Humans; Immunity; Immunoglobulin E; Immunomodulation; Inflammation Mediators; Interleukin-8; Molecular Weight; Nasal Mucosa; Nasal Provocation Tests; Plant Extracts; Pollen; Rhinitis, Allergic, Seasonal; Skin Tests; Symptom Assessment; Th2 Cells

Allergen-specific immunotherapy (IT) is widely used to treat allergic diseases. The molecular mechanisms have not been clarified yet completely. The present work was undertaken to analyze the effect of IT in the activation of NF-κB.. Neutrophils from 15 pollen-allergic IT-treated patients, 10 untreated pollen-allergic patients, and 10 healthy donors were in vitro stimulated with LPS. NF-κB activation (p65/p52) was measured in their nuclear extracts by enzyme-linked immunosorbent assay (ELISA). IκBα phosphorylation, NF-κB-repressing factor (NRF) activation, and thromboxane A2 (TXA2 ) and Interleukin-8 (IL-8) release were measured by ELISA.. There was a positive correlation between the score of symptoms and NF-κB activation in human neutrophils. IT significantly decreased NF-κB activation levels in neutrophils compared with neutrophils from untreated patients. IκBα phosphorylation and NRF activation levels were, respectively, significantly lower and higher in neutrophils from IT-treated patients than from untreated patients. IL-8 and TXA2 release were significantly lower in neutrophils from IT-treated patients than from untreated patients.. IT positive effects are at least in part mediated by the negative regulation of NF-κB activation in human neutrophils. These observations represent a novel view of neutrophils as possible cell target to treat IgE-dependent diseases through NF-κB downmodulation.

Topics: Adolescent; Allergens; Case-Control Studies; Cells, Cultured; Dactylis; Desensitization, Immunologic; Down-Regulation; Female; Humans; I-kappa B Proteins; Inflammation Mediators; Interleukin-8; Lipopolysaccharides; Male; Neutrophils; NF-kappa B; NF-kappa B p52 Subunit; NF-KappaB Inhibitor alpha; Phosphorylation; Pollen; Repressor Proteins; Rhinitis, Allergic, Seasonal; Signal Transduction; Thromboxane A2; Transcription Factor RelA; Treatment Outcome

For locally acting drugs, an extended residence time in the nasal cavity is desirable and related to a prolonged effect. We sought to develop a model for comparative determination of intranasal pharmacokinetics. We embedded human respiratory tissue into a solid matrix and coated the surface with artificial nasal fluid. Nasal spray suspensions of fluticasone propionate (FP) and budesonide (Bud) as well as a solution of azelastine hydrochloride (AZ) were applied onto the surface and removed after 30 min to simulate mucociliary clearance. As exemplary anti-inflammatory measure, we evaluated the inhibition of IL-8 release from epithelial cells. FP and Bud were initially bound to the same extent to the tissue gel while AZ displayed a more 4-fold higher binding than FP or Bud. After equilibrium with plasma, approximately 5-fold higher tissue concentrations of AZ compared to FP and 77-fold higher levels in relation to Bud were determined. This tissue retention revealed an excellent correlation with the volume of distribution of the respective drugs (r=0.9999, p ≤ 0.05). The inhibitory effect of FP on IL-8 release was approximately 5-fold more pronounced compared to AZ. The present model realistically mirrors conditions in vivo where solubility and tissue absorption of intranasally applied drugs compete with mucociliary clearance mechanisms.

Topics: Adenocarcinoma; Adenocarcinoma of Lung; Administration, Inhalation; Administration, Intranasal; Aerosols; Androstadienes; Anti-Allergic Agents; Anti-Inflammatory Agents; Budesonide; Cell Line, Tumor; Delayed-Action Preparations; Epithelial Cells; Fluticasone; Glucocorticoids; Histamine Antagonists; Humans; Interleukin-8; Lung; Lung Neoplasms; Mucociliary Clearance; Nasal Lavage Fluid; Nasal Sprays; Phthalazines; Rhinitis, Allergic, Seasonal

Natural killer cells (NK) secrete eosinophilotactic cytokines, however, whether they contribute to eosinophil chemotaxis by secreting IL-8 is not known. We investigated the ability of CD56+CD3-ve (NK cells) to induce chemotaxis of peripheral blood eosinophils from allergic rhinitis (AR) patients, through IL-8 secretion, and the effects of IL-15, the NK cell proactivating cytokine, and calcitriol: 1α, 25-dihydroxy Vitamin D(3) (vitamin D(3)), the immunomodulator agent, in this scenario. Herein, it is shown that supernatants from unstimulated NK cells exhibited chemotactic activity against eosinophil. This effect was significantly augmented by IL-15 (1 ng/mL) treatment, resulting in an increase in the chemotactic index of approximately 3 folds and was abrogated by neutralizing antibody (Ab) to IL-8 in a dose-dependent fashion. The amount of IL-8 secreted by NK cells was increased by IL-15 treatment from levels of 88.64 ± 21.5 to 178.9 ± 23.6 Pg/mL and was significantly reduced by 10(-6) M vitamin D(3) to levels of 59.2 ± 16.3 Pg/mL. Our results indicate a novel inflammatory crosstalk between NK cells and eosinophils via IL-15/IL-8 axis that can be modulated by vitamin D(3).

Topics: Antibodies, Neutralizing; Autoantibodies; Autocrine Communication; Cell Communication; Cells, Cultured; Chemotaxis; Cholecalciferol; Eosinophils; Humans; Interleukin-15; Interleukin-8; Killer Cells, Natural; Receptor Cross-Talk; Rhinitis, Allergic, Seasonal

Viral respiratory infections are increasingly implicated in allergic exacerbations. Virus-induced activation of eosinophils through Toll-like receptors (TLRs) could be involved. The present study was designed to examine TLR3 expression in eosinophils from bone marrow (BM) and peripheral blood (PB) during symptomatic allergic rhinitis, and to evaluate the functional responsiveness of TLR3 in purified eosinophils.. BM and PB samples were obtained from healthy volunteers and patients with seasonal allergic rhinitis outside and during the pollen season. Eosinophils were analyzed for TLR3 expression by flow cytometry. Polyinosinic:polycytidylic acid [poly(I:C)], an agonist for TLR3, was used to assess its functional role in purified eosinophils and the intracellular signaling pathways involved.. TLR3 expression was demonstrated in BM and PB eosinophils. It was higher in BM-derived than in circulating cells and it was downregulated in both compartments during symptomatic allergic rhinitis. TLR3 expression was also downregulated in the presence of interleukin (IL)-4 and IL- 5. Stimulation with poly(I:C) increased the percentage of CD11b+ cells and enhanced the secretion of IL-8, effects mediated via the p38 mitogen-activated protein kinases and nuclear factor-kappaB signaling pathways. Moreover, pretreatment with IL-5 augmented the poly(I:C)-induced IL-8 release.. Eosinophils activated via TLR3 might be more able to home and recruit leukocytes to sites of inflammation. The decreased TLR3 expression during symptomatic allergic rhinitis and in the presence of Th2 cytokines indicates a role in allergic airway inflammation. Thus, eosinophils might function as a link between viral infections and exacerbations of allergic disease.

Topics: Adult; Blood Cell Count; Bone Marrow Cells; CD11b Antigen; Cell Count; Cysteine Proteinase Inhibitors; Eosinophils; Female; Gene Expression; Humans; Imidazoles; Interleukin-4; Interleukin-5; Interleukin-8; Leupeptins; Male; Middle Aged; Neutrophils; NF-kappa B; p38 Mitogen-Activated Protein Kinases; Phosphorylation; Poly I-C; Protein Kinase Inhibitors; Pyridines; Rhinitis, Allergic, Seasonal; Signal Transduction; Toll-Like Receptor 3; Virus Diseases; Young Adult

Among proteases, metalloproteases are implicated in tissue remodeling, as shown in numerous diseases including allergy. ADAMs (A Disintegrin And Metalloprotease) metalloproteases are implicated in physiologic processes such as cytokine and growth factor shedding, cell migration, adhesion, or repulsion. Our aim was to measure ADAM-12 expression in airway epithelium and to define its role during the allergic response. To raise this question, we analyzed the ADAM-12 expression ex vivo after allergen exposure in patients with allergic rhinitis and in vitro in cultured primary human airway epithelial cells (AEC). Clones of BEAS-2B cells transfected with the full-length form of ADAM-12 were generated to study the consequences of ADAM-12 up-regulation on AEC function. After allergen challenge, a strong increase of ADAM-12 expression was observed in airway epithelium from patients with allergic rhinitis but not from control subjects. In contrast with the other HB-epidermal growth factor sheddases, ADAM-10 and -17, TNF-alpha in vitro increased the expression of ADAM-12 by AEC, an effect amplified by IL-4 and IL-13. Up-regulation of ADAM-12 in AEC increased the expression of alpha3 and alpha4 integrins and to the modulation of cell migration on fibronectin but not on collagen. Moreover, overexpression of ADAM-12 in BEAS-2B enhanced the secretion of CXCL1 and CXCL8 and their capacity to recruit neutrophils. CD47 was strongly decreased by ADAM-12 overexpression, a process associated with a reduced adhesion of neutrophils. These effects were mainly dependent on epidermal growth factor receptor activation. In summary, ADAM-12 is produced during allergic reaction by AEC and might increase neutrophil recruitment within airway mucosa.

Topics: ADAM Proteins; ADAM12 Protein; Allergens; Bronchi; CD47 Antigen; Cell Adhesion; Cells, Cultured; Chemokine CXCL1; Chemotaxis, Leukocyte; Epithelial Cells; ErbB Receptors; Gene Expression Regulation; Humans; Integrins; Interleukin-8; Membrane Proteins; Neutrophils; Recombinant Fusion Proteins; Rhinitis, Allergic, Perennial; Rhinitis, Allergic, Seasonal; Transfection; Tumor Necrosis Factor-alpha

TNFalpha is a cytokine that may contribute to the pathophysiology of airway inflammation. Inhalation of TNFalpha produces granulocyte recruitment and airway hyperresponsiveness in man. Anti-TNFalpha treatment may inhibit allergen-induced plasma exudation in guinea-pig airways. Increased nasal mucosal output of TNFalpha has been demonstrated in allergic rhinitis, but the effect of TNFalpha on the human nasal mucosa has not been examined in vivo.. To examine effects of topical TNFalpha on the human nasal mucosa in vivo.. In a dose-finding study, healthy subjects received intranasal TNFalpha (0-7.5 microg). Nasal lavages were carried out before as well as 10 min and 24 h post challenge and alpha(2)-macroglobulin was measured as an index of plasma exudation. In a second study, involving patients with allergic rhinitis examined out of season, a sham-controlled nasal challenge with TNFalpha (10 microg) was performed and followed 24 h later by an allergen challenge. Lavages were performed before the TNFalpha challenge, 24 h thereafter, and 10 min post allergen challenge. alpha(2)-Macroglobulin, eosinophil cationic protein (ECP), myeloperoxidase (MPO), and IL-8 were analyzed as indices of plasma exudation, eosinophil activity, neutrophil activity, and pro-inflammatory cytokine production, respectively.. In the dose-finding study, TNFalpha produced significant increases in alpha(2)-macroglobulin 24h post challenge (p<0.01). In allergic rhinitis, 10 microg of TNFalpha also produced this effect (p<0.01) as well as increases in ECP and IL-8 (p<0.01). MPO was increased 24 h post challenge, but this change did not reach statistical significance. TNFalpha did not produce any acute effects and did not affect the responsiveness to allergen.. The present study demonstrates that topical TNFalpha produces a human nasal inflammatory response. These data suggest a role of TNFalpha in nasal conditions characterized by mucosal inflammation.

Topics: Adolescent; Adult; Allergens; alpha-Macroglobulins; Dose-Response Relationship, Immunologic; Eosinophil Cationic Protein; Female; Humans; Inflammation Mediators; Interleukin-8; Male; Nasal Lavage Fluid; Nasal Mucosa; Peroxidase; Rhinitis, Allergic, Seasonal; Tumor Necrosis Factor-alpha

To survey the effects of TNF-alpha and IL-8 in the pathogenesis of allergic rhinitis.. The serum levels of TNF-alpha and IL-8 in 102 cases of seasonal or non-seasonal allergic rhinitis were detected pre and post specific immunotherapy (SIT), and the results were statistically analyzed.. The levels of TNF-alpha and IL-8 in experiment group were significant higher than that in the health control. And the levels of TNF-alpha and IL-8 significantly decreased post SIT in the experiment group.. TNF-alpha and IL-8 play important roles in the pathogenesis of allergic rhinitis.

Topics: Adolescent; Adult; Humans; Interleukin-8; Middle Aged; Rhinitis, Allergic, Perennial; Rhinitis, Allergic, Seasonal; Tumor Necrosis Factor-alpha; Young Adult

Allergic rhinitis is characterized by a T(H)2-dependent inflammation. Nasal obstruction is a typical symptom of allergic rhinitis.. To evaluate the possible relationships among nasal symptoms, allergic inflammation, including inflammatory cells and cytokine pattern, and nasal airflow in children with seasonal allergic rhinitis.. Children with seasonal allergic rhinitis and moderate-severe nasal obstruction were evaluated during the pollen season. Total symptom score, rhinomanometry, nasal lavage, and nasal scraping were evaluated in all patients. Inflammatory cells were counted by conventional staining; interleukin 5 (IL-5) and IL-8 levels were measured by immunoassay on fluids recovered from nasal lavage.. Twenty children (11 boys and 9 girls; mean +/- SD age, 12.9 +/- 1.7 years) participated in this study. Eosinophil levels were significantly associated with total symptom score (r = 90.6%, P < .001), IL-5 (r = 94.9%, P < .001), and nasal flow (r = -93.6%, P < .001). No association was elicited with IL-8 (r = 9.4%, P = .69). In a multivariate analysis that included eosinophils, neutrophils, and IL-5, eosinophil levels were shown to be the only independent predictor of nasal flow.. This study demonstrates the close connection between T(H)2 cytokines and eosinophil infiltration. In addition, there is clear evidence concerning the relationship among nasal symptoms, eosinophil infiltration, and nasal airflow. These findings constitute evidence of the relationship between nasal airflow impairment and eosinophilic inflammation in children with seasonal allergic rhinitis.

Topics: Child; Eosinophils; Female; Humans; Hypersensitivity; Inflammation; Interleukin-5; Interleukin-8; Male; Nasal Lavage Fluid; Nasal Obstruction; Nose; Pulmonary Ventilation; Rhinitis, Allergic, Seasonal; Skin Tests; Th2 Cells

Allergic rhinitis is characterized by a Th2-dependent inflammation. Nasal obstruction largely depends on allergic inflammation.. The aim of this study was to evaluate the possible role of the symptom nasal obstruction in assessing patients with hay fever.. Fifty patients (mean age, 23.7 +/- 4.9 years) with hay fever were evaluated both during and outside pollen season. All of them had moderate-severe grade of nasal obstruction. Total symptom score (TSS), rhinomanometry, nasal lavage, nasal scraping, spirometry, and methacholine bronchial challenge were performed in all subjects.. During the pollen season, patients with severe nasal obstruction showed significantly higher values of TSS, IL-4, IL-5, IL-8, nasal eosinophils and neutrophils, and significantly lower values of nasal airflow, IFNgamma, FEV1, FVC, and FEF 25-75 in comparison with patients with moderate nasal obstruction. Twenty (83%) patients with severe nasal obstruction showed bronchial hyperreactivity (BHR), whereas only 6 (25%) patients with moderate nasal obstruction had BHR. Outside the pollen season overlapping results were observed.. This study provides evidence about the key role played by nasal obstruction in assessing patients with allergic rhinitis.

Topics: Adult; Enzyme-Linked Immunosorbent Assay; Eosinophils; Humans; Interferon-gamma; Interleukin-4; Interleukin-5; Interleukin-8; Male; Nasal Lavage Fluid; Nasal Obstruction; Neutrophils; Pollen; Pyroglyphidae; Rhinitis, Allergic, Seasonal; Rhinomanometry; Severity of Illness Index; Skin Tests; Spirometry; Th2 Cells

Eosinophils are seen together with neutrophils at sites of inflammation. However, their roles are not clear. In addition, eosinophils infiltrate tumor tissue in some neoplastic diseases. In this study, we show that large amounts of the neutrophil-activating CXC chemokine growth-related oncogene (GRO)-alpha can be produced by human eosinophils. Eosinophils showed presence of preformed GRO-alpha in the crystalloid-containing specific granules (190 pg/2 x 10(6) cells). During incubation, a strong increase in GRO-alpha gene expression was seen. At a low cell density, addition of TNF-alpha or IL-1 beta increased the production of GRO-alpha in eosinophils, which was not the case at a higher cell density. Eosinophils can produce TNF-alpha themselves, and neutralizing Abs against TNF-alpha significantly inhibited GRO-alpha production. This suggests that autocrine and paracrine effects from TNF-alpha can be important when up-regulating GRO-alpha gene expression. In contrast, IFN-gamma, a prototypic Th1-cytokine, down-regulated expression of GRO-alpha. This may be important during resolution of inflammation but also suggests different roles for eosinophils depending on the inflammatory context. Tumor-infiltrating eosinophils in Hodgkin's disease of the nodular sclerosing type are associated with a poor prognosis. Eosinophils from such tumor tissue showed an abundant expression of GRO-alpha. The GRO-alpha receptor CXCR2 was also detected in tumor tissue, proposing interactions between eosinophils and the tumor. Our findings suggest that eosinophils can promote inflammation through recruitment of CXCR2-bearing cells. In addition, this feature of the eosinophils indicates a role for these cells in the biology of certain tumors.

Topics: Autocrine Communication; Cell Communication; Chemokine CXCL1; Chemokines; Chemokines, CXC; Chemotactic Factors; Down-Regulation; Eosinophils; Hodgkin Disease; Humans; Immunohistochemistry; Intercellular Signaling Peptides and Proteins; Interferon-gamma; Interleukin-5; Interleukin-8; Microscopy, Immunoelectron; Neoplasm Proteins; Paracrine Communication; Rhinitis, Allergic, Seasonal; Time Factors; Tumor Necrosis Factor-alpha; Up-Regulation

Myeloperoxidase (MPO) has been proposed to mirror the degree of neutrophil activation, however its role as a marker of the participation of neutrophils in allergic inflammation is still unclear as the literature is controversial. The aim of this study was to evaluate the MPO levels and neutrophil influx in nasal lavage after recombinant Il-8 challenge. Eight patients suffering from seasonal allergic rhinitis, hypersensitive to grass pollens, with average age 30.1 +/- 2.67 were challenged both with Il-8 and diluent for Il-8 on a subsequent day, in two phases: before the pollen season (unprimed mucosa) and during the season (primed mucosa). Nasal lavage with saline were collected before, during Il-8 or placebo challenge and 30 minutes, 2 hours and 3 hours after the challenge. The number of neutrophils and MPO levels in the nasal fluid were determined. After the challenge with Il-8 of primed mucosa the number of neutrophils increased from 28250 cells/ml at the baseline to 28778, 251020 and 333660 at 0, 5, 2 and 3 hours respectively. There was a statistically significant difference between cells number after diluent and Il-8 challenge (p < 0.05, Wilcoxon rank-sum test). The number of neutrophils in the nasal lavage of primed mucosa after Il-8 challenge was significantly higher (p < 0.005) at all time points in comparison with the diluent and Il-8 challenges in the unprimed mucosa. There was no difference (p < 0.05) in MPO levels in the nasal lavage between Il-8 (mean 17.43 ng/ml +/- 10.98) and diluent challenge (20.98 ng/ml +/- 11.89) of unprimed mucosa. In the primed mucosa fluid we observed a peak of MPO level at 2 hours time point, however that was not significant as compared to diluent challenge (p = 0.465). We did not find the relationship between MPO levels and the neutrophils number in the lavage (rank Spearman factor, rs = 0.258, p = 0.42). Due to the lack of statistically confirmed relationship between MPO level and the number of neutrophils, MPO seems to be of little value as a marker of neutrophil influx into nasal mucosa.

Topics: Adult; Allergens; Biomarkers; Data Interpretation, Statistical; Dose-Response Relationship, Drug; Female; Humans; Interleukin-8; Male; Nasal Lavage Fluid; Nasal Mucosa; Nasal Provocation Tests; Neutrophils; Peroxidase; Rhinitis, Allergic, Seasonal; Time Factors

Allergic inflammation is regulated by the local production and release of several cytokines.. This study was designed to assess the changes in mRNA cytokine-positive cells after allergen provocation and to compare these cytokines with tissue eosinophilia as a marker of allergic inflammation.. A grass pollen allergen provocation study was conducted in autumn, out of the hay fever season. Nasal mucosal biopsy specimens were taken before provocation and 1 hour, 24 hours, and 1 week after allergen provocation. Eosinophils and mRNA-positive cells (in situ hybridization for IL-2, IL-3, IL-4, IL-5, IL-6, IL-8, IL-10, IL-13, IFN-gamma, RANTES, and TNF-alpha) were assessed in the biopsy specimens.. After allergen provocation, an increase in cell number was found for eosinophils and cells expressing mRNA for the chemokines IL-8 and RANTES and for the TH2 cytokines IL-10 and IL-13. Significant correlations were found between eosinophils and RANTES and eosinophils and IFN-gamma in the early phase and between eosinophils and IL-5 and eosinophils and RANTES in the late phase. The increase in eosinophils and IL-10 and IL-13 mRNA-positive cells could still be observed 1 week after allergen provocation.. Nasal allergen provocation induced significant tissue eosinophilia and a significant increase in IL-8, IL-13, and RANTES mRNA-positive cells. A significant increase in eosinophils and IL-10 and IL-13 mRNA-positive cells compared with baseline can still be observed 1 week after a single allergen provocation.

Topics: Adult; Allergens; Biopsy; Cell Count; Chemokine CCL5; Eosinophilia; Eosinophils; Female; Humans; In Situ Hybridization; Interferon-gamma; Interleukin-10; Interleukin-13; Interleukin-8; Interleukins; Male; Middle Aged; Nasal Mucosa; Nasal Provocation Tests; Poaceae; Pollen; Rhinitis, Allergic, Seasonal; RNA, Messenger; Seasons; Tumor Necrosis Factor-alpha

The objectives of this study were to measure interleukins 5 and 8 (IL-5 and IL-8) in relation to eosinophils and neutrophils, in nasal lavage fluids from 60 school children with allergic rhinitis, and to determine the influence of treatment with a topical steroid (budesonide) on the levels of the two cytokines. Highly sensitive enzyme immunoassays were used to analyze IL-5 and IL-8. IL-5 levels and relative eosinophil counts in nasal lavage fluid increased significantly in patients with allergic rhinitis during the pollen season, compared with values obtained before the start of the season, and decreased significantly after treatment with budesonide. By contrast, no significant changes in IL-8 or neutrophils were found during the pollen season, nor did they decrease following treatment. In the untreated patients, IL-5 levels correlated significantly with eosinophil counts but not with neutrophil counts, whereas IL-8 levels correlated with neutrophil counts but not with eosinophil counts. After budesonide treatment, the correlation between IL-8 and neutrophils remained, and a correlation between IL-8 and eosinophils emerged. These findings support the concepts that IL-5 has a key role in regulating eosinophils and that IL-8 is important for the regulation of neutrophils. Whereas IL-5 and relative eosinophil counts are profoundly affected by topical steroid treatment, IL-8 and neutrophils are not demonstrably affected by such treatment. It is possible that neutrophils, through the release of IL-8, could be chemotactic for eosinophils in steroid-treated patients.

Topics: Adolescent; Adult; Allergens; Anti-Inflammatory Agents; Budesonide; Child; Enzyme-Linked Immunosorbent Assay; Eosinophils; Humans; Interleukin-5; Interleukin-8; Leukocyte Count; Nasal Lavage Fluid; Neutrophils; Pollen; Prospective Studies; Rhinitis, Allergic, Seasonal

Recent studies have suggested that antihistamines, widely used in the treatment of symptoms of patients with allergic rhinitis, may also possess antiinflammatory properties. The mechanisms underlying this property, however, are not clearly understood. We have cultured epithelial cells from nasal biopsy specimens from patients with seasonal allergic rhinitis outside the pollen season and studied the effect of 0 to 10(-3) mol/L fexofenadine, the main active metabolite of terfenadine, on eosinophil-induced changes in electrical resistance (measure of permeability) and release of proinflammatory mediators from these cells. Additionally, we have studied the effect of this drug on eosinophil chemotaxis and adherence to endothelial cells induced by conditioned medium from these human nasal epithelial cell (HNEC) cultures. Incubation of HNEC in the presence of eosinophils treated with opsonized latex beads significantly decreased the electrical resistance of these cultures, an effect that was abrogated by treatment of the cultures with 10(-9) to 10(-3) mol/L fexofenadine. Similarly, incubation of HNEC in the presence of eosinophils treated with latex beads also significantly increased the basal release of the chemokine "regulated upon activation, normal T cell expressed and secreted" (RANTES) (from 96.0 to 613.0 fg/microg cellular protein; p < 0.05), IL-8 (from 42.0 to 198.5 pg/microg cellular protein; p < 0.05), granulocyte-macrophage colony-stimulating factor (GM-CSF) (from 0.54 to 3.4 pg/microg cellular protein; p < 0.05), and soluble intercellular adhesion molecule-1 (sICAM-1) (from 7.8 to 18.4 pg/microg cellular protein; p < 0.05) from HNEC. The eosinophil-induced release of IL-8, GM-CSF, and sICAM-1 from the HNEC was significantly attenuated by treatment with fexofenadine. Analysis of the effects of conditioned medium from HNEC demonstrated that this significantly increased both eosinophil chemotaxis and adherence to endothelial cells. Addition of 10(-6) to 10(-3) mol/L fexofenadine to the conditioned medium significantly attenuated eosinophil chemotaxis and adherence to endothelial cells. These results suggest that fexofenadine may reduce nasal inflammation by modulating the release of proinflammatory mediators and adhesion molecules from HNEC.

Topics: Cell Adhesion; Cell Membrane Permeability; Cells, Cultured; Chemokine CCL5; Chemotaxis, Leukocyte; Culture Media, Conditioned; Cytokines; Endothelium, Vascular; Eosinophils; Epithelial Cells; Granulocyte-Macrophage Colony-Stimulating Factor; Histamine Antagonists; Humans; Intercellular Adhesion Molecule-1; Interleukin-8; Microspheres; Nasal Mucosa; Opsonin Proteins; Proteins; Rhinitis, Allergic, Seasonal; T-Lymphocytes; Terfenadine

Leukotriene B4 (LTB4), a potent chemokinetic mediator for neutrophils, is enhanced by interleukin-8 (IL-8) and may play a key role in the inflammatory response of asthma.. The aim of the present study was to investigate whether zileuton, a 5-lipoxygenase antagonist known to inhibit LTB4 production and recruitment of eosinophils/neutrophils in bronchoalveolar fluid, could affect the production of LTB4 and IL-8 by allergen-stimulated peripheral blood mononuclear cells in vitro from patients with asthma and/or allergic rhinitis.. Peripheral blood mononuclear cells were isolated using Ficoll-Hypaque density gradient from 14 subjects (2 with asthma, 11 with asthma and allergic rhinitis, and 1 with allergic rhinitis) and were stimulated by selected allergens (grass, tree, mite, and mold) in the absence or presence of 1 and 10 microM of zileuton. Supernatants were collected and assayed for LTB4 and IL-8 levels using RIA and ELISA, respectively.. Levels of LTB4 were significantly elevated in peripheral blood mononuclear cells stimulated with mold, grass, and tree compared with the unstimulated control group (P<.05). Levels of IL-8 were significantly elevated in all allergen-stimulated peripheral blood mononuclear cells, except mold, compared with the unstimulated control group (P<.05). Zileuton significantly reduced production of LTB4 by mold and tree-stimulated peripheral blood mononuclear cells. By contrast, no effect of zileuton on IL-8 production was observed in allergen-stimulated peripheral blood mononuclear cells.. The zileuton-induced attenuation of LTB4 production by allergen-stimulated peripheral blood mononuclear cells from patients with asthma and/or allergic rhinitis occurs independently from the allergen-stimulated IL-8 production.

Topics: Adult; Aged; Allergens; Asthma; Female; Humans; Hydroxyurea; Interleukin-8; Leukocytes, Mononuclear; Leukotriene B4; Lipoxygenase Inhibitors; Male; Middle Aged; Rhinitis, Allergic, Perennial; Rhinitis, Allergic, Seasonal

Recent studies have suggested that airway epithelial cells of atopic and nonatopic individuals may differ in their ability to produce proinflammatory cytokines.. We have cultured human nasal epithelial cells (NECs) as confluent explant cultures from nasal biopsy specimens of well-characterized nonatopic normal volunteers without rhinitis (n = 8), atopic volunteers without rhinitis (n = 9), and atopic patient volunteers with rhinitis (n = 10) and measured the amounts of IL-1 beta, IL-8, granulocyte-macrophage colony-stimulating factor, tumor necrosis factor-alpha, and RANTES released spontaneously into the culture medium by these cells in vitro. NECs from patients with allergic rhinitis were cultured from biopsy specimens obtained on two different occasions, during and after the pollen season.. In general, NECs from atopic individuals released significantly greater amounts of IL-1 beta, IL-8, granulocyte-macrophage colony-stimulating factor, tumor necrosis factor-alpha, and RANTES than NECs from nonatopic individuals. IL-8 was released in greatest quantity and IL-1 beta in lowest quantity, regardless of whether the NECs were derived from atopic or nonatopic volunteers. Of the atopic individuals, NECs of atopic patients with rhinitis naturally exposed to pollen released greater quantities of all these cytokines, compared with NECs of atopic patients with rhinitis and atopic patients without rhinitis who were not exposed to allergen.. These results suggest that NECs of atopic individuals, who are genetically predisposed to upper airway disease, release increased amounts of proinflammatory cytokines and that natural exposure to allergen enhances the release of these cytokines, exacerbating the symptoms of allergic disease.

Topics: Adolescent; Adult; Biopsy; Cells, Cultured; Chemokine CCL5; Culture Media; Cytokines; Epithelial Cells; Epithelium; Female; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Hypersensitivity, Immediate; Interleukin-1; Interleukin-8; Male; Nasal Mucosa; Rhinitis, Allergic, Seasonal; Tumor Necrosis Factor-alpha

Interleukin-8 (IL-8) is a chemotactic cytokine for neutrophils and primed eosinophils. In allergic rhinitis, allergen exposure triggers leucocyte recruitment.. We evaluated in this study IL-8 secretion and the neutrophil chemotactic activity in nasal lavages collected after a nasal allergen challenge. Moreover, the participation of IL-8 in the neutrophil chemotactic activity was quantified.. Four healthy subjects and 19 patients with allergic rhinitis were exposed to a nasal allergen challenge. As a control, saline challenge was performed in four patients with allergic rhinitis. Concentration of IL-8 was measured by ELISA in nasal lavages collected before and after challenge. Neutrophil chemotactic assay was developed using a 48-well chemotaxis microassembly.. After allergen challenge, the healthy subjects, the four patients receiving saline and one patient exposed to allergen did not respond; seven patients presented a single early reaction and 11 patients a dual response. For healthy subjects and the four patients exposed to saline, the level of IL-8 did not increase after challenge in comparison with that at baseline. After allergen challenge, two peaks of IL-8 release were observed for patients with allergic rhinitis during the early (30 min to 1 h 30 min) and the late periods (3 h 30 min to 9 h 30 min), however the difference was not significant for the early period. During the late period, a significant increase in IL-8 concentrations was detected for the patients developing a dual response, whereas the difference was not significant for those presenting only an early reaction. The neutrophil chemotactic activity of nasal lavages from patients with allergic rhinitis collected during the early and the late reactions (17 +/- 2.1 and 23.3 +/- 2.8 neutrophils per high power field (hpf), respectively) was significantly higher than the activity of lavage fluid collected at baseline (9.2 +/- 1.8 neutrophils per hpf). Nevertheless, the addition of a neutralizing anti-IL-8 antibody inhibited weakly the chemotactic activity of lavage fluid from rhinitic patients collected during the early or the late periods (18 and 11% of inhibition) (P = NS).. These data show that allergen challenge increased significantly the secretion of IL-8 for the patients with allergic rhinitis. However, neutralization of IL-8 in nasal lavages by a specific antibody revealed that the role of this chemokine in granulocyte infiltrate was limited, suggesting that IL-8 acts in connection with other chemotactic factors in this recruitment.

Topics: Adult; Allergens; Animals; Case-Control Studies; Chemotactic Factors; Chemotaxis, Leukocyte; Female; Humans; In Vitro Techniques; Interleukin-8; Male; Mites; Nasal Lavage Fluid; Nasal Mucosa; Nasal Provocation Tests; Neutralization Tests; Neutrophils; Pollen; Rhinitis, Allergic, Perennial; Rhinitis, Allergic, Seasonal

Monocyte chemotactic and activating factor/monocyte chemoattractant protein-1 (MCAF/MCP-1), RANTES, and macrophage inflammatory protein (MIP)-1 alpha are chemokines known to activate basophils (MCAF/RANTES) and eosinophils (RANTES/MIP-1 alpha). IL-8 inhibits MCAF-induced histamine release from basophils. We questioned whether a relationship exists between the levels of these chemokines and various inflammatory mediators released from mast cells, eosinophils, and basophils as assessed in nasal secretions obtained from patients during the allergy season and out of season. Samples were assessed for MCAF/MCP-1, RANTES, MIP-1 alpha, IL-8, histamine, tryptase and eosinophil cationic protein (ECP) in three subject groups: subjects with allergic rhinitis (n = 18), atopic subjects without rhinitis (n = 9), and healthy individuals (n = 6). Statistically significant differences were apparent only in the subjects with symptoms as follows. MCAF/MCP-1 increased during the season from 336 +/- 47 pg/ml to 829 +/- 137 pg/ml (p < 0.001), whereas IL-8 decreased from a baseline of 1932 +/- 335 pg/ml to 1070 +/- 202 pg/ml (p < 0.028). The ratio of IL-8 to MCAF/MCP-1 decreased during the symptomatic season from the baseline of 6.66 +/- 1.06 seen during winter to 1.3 +/- 0.22 during ragweed season (p < 0.001). Histamine increased from 6.3 +/- 1.5 to 89 +/- 15.5 ng/ml (p < 0.001), ECP increased from 20.6 +/- 6.4 to 237.1 +/- 50.2 ng/ml (p < 0.001), and tryptase increased from 2.34 +/- 0.6 to 9.7 +/- 2.3 U/ml (p < 0.001). Most samples did not have detectable quantities of MIP-1 alpha or RANTES. We also found a correlation between the level of MCAF/MCP-1 and IL-8 and the level of histamine or IL-8 and ECP. Our results suggest that the chemokines MCAF/MCP-1 and IL-8 may participate in the pathogenesis of allergic rhinitis, contributing to the attraction of the proinflammatory cells and mediator release, which might be very important during the late phase of the allergic reaction. Furthermore, the ratio of certain chemokines, such as MCAF/MCP-1 and IL-8 may reflect the magnitude of the reaction, as does the presence of histamine and ECP.

Topics: Basophils; Blood Proteins; Chemokines; Chymases; Eosinophil Granule Proteins; Eosinophils; Histamine; Humans; Interleukin-8; Nasal Mucosa; Rhinitis, Allergic, Seasonal; Ribonucleases; Serine Endopeptidases; Tryptases

Epithelial cells potentially contribute to airways inflammation by antigen presentation and the production of proinflammatory cytokines. This study investigated the immunocytochemical localization of interleukin-8 (IL-8), granulocyte-macrophage colony-stimulating factor (GM-CSF), interleukin-1 receptor (IL-1R Type I), tumor necrosis factor-alpha receptor (TNF-alpha R; 55kD), and human leukocyte antigen-DR (HLA-DR) on epithelial cells obtained by nasal brushing from 10 patients with allergic rhinitis in season and 15 healthy, nonallergic subjects. Six of the 15 nonallergic asymptomatic subjects had macroscopic evidence of nasal mucosal inflammation, and their brushings contained more than 10% neutrophils ("subclinical inflammation"). In normal control subjects, 8 +/- 7.5% of epithelial cells stained for HLA-DR, approximately one quarter stained for IL-8 and GM-CSF, and about one third stained positive for IL-1R and TNF-alpha R. The findings in subjects with allergic rhinitis in season and with subclinical neutrophilia were similar, and the numbers of cells staining for HLA-DR expression correlated with both neutrophil and lymphocyte content. These findings further support the conclusion that epithelial cells can contribute to inflammatory processes in the nasal mucosa. The findings emphasize the need to identify asymptomatic nasal mucosal inflammation in studies of the nasal mucosa.

Topics: Adolescent; Adult; Eosinophils; Epithelium; Granulocyte-Macrophage Colony-Stimulating Factor; HLA-DR Antigens; Humans; Interleukin-8; Keratins; Leukocyte Count; Macrophages; Middle Aged; Nasal Mucosa; Neutrophils; Receptors, Interleukin-1; Receptors, Tumor Necrosis Factor; Rhinitis; Rhinitis, Allergic, Seasonal; T-Lymphocytes, Helper-Inducer; Tumor Necrosis Factor-alpha

Allergic diseases such as allergen-induced rhinitis represent an inflammatory reaction that is characterized by the chemotaxis and activation of various cell populations. A high degree of cell-to-cell communication is needed to orchestrate this inflammatory immune response. A variety of cytokines and adhesion receptors seem to play an important role in the allergic late phase reaction. Here we demonstrate that proinflammatory cytokines such as interleukin(IL)-1, IL-8 and TNF-alpha (tumor necrosis factor-alpha) can be detected in nasal secretions and mucosa by enzyme-linked immunosorbent assay and immunohistochemistry. The increased expression of adhesion receptors in mucosa specimens of patients with seasonal allergic rhinitis points to their role in regulating the cellular migration and probably represents a key event in allergic inflammation. We established an in vitro model using freshly taken nasal mucosa to study the induction of adhesion receptors by proinflammatory cytokines. E-selectin, an endothelial receptor, was strongly upregulated by IL-1 beta, TNF-alpha and allergen. The induction due to allergen exposure of the mucosa was markedly inhibited by soluble cytokine receptors (sIL-1R, TNF-BP) or by a receptor antagonist (IL-1ra) and prednisolone, These findings indicate that proinflammatory cytokines may be key factors for the upregulation of adhesion processes in human nasal mucosa and the activation of various cell populations involved in the allergic inflammation. They therefore represent a main target for new therapeutic strategies.

Topics: Cell Adhesion Molecules; Cell Communication; Chemotaxis; E-Selectin; Humans; Intercellular Adhesion Molecule-1; Interleukin-1; Interleukin-8; Lymphocyte Function-Associated Antigen-1; Membrane Glycoproteins; Nasal Mucosa; Receptors, Immunologic; Respiratory Hypersensitivity; Rhinitis; Rhinitis, Allergic, Seasonal; Tumor Necrosis Factor-alpha; Vascular Cell Adhesion Molecule-1

Topics: Animals; Cathepsin G; Cathepsins; Cell Adhesion; Cell Degranulation; Cell Movement; Dogs; Interleukin-8; Muramidase; Neutrophils; Pancreatic Elastase; Rhinitis, Allergic, Seasonal; Serine Endopeptidases; Trachea