Fluticasone Proprionate Research Articles

Conservative treatment of phimosis with fluticasone proprionate 0.05%: A clinical study in 1185 boys

Circumcision has been the traditional method of choice in the treatment of boys with phimosis. Recently, several published studies worldwide have focused their interest on more conservative approaches in management of this condition. These studies advocate the use of topical steroids in the phimotic foreskin. We aimed to investigate the efficacy and safety of a medium potency corticosteroid in boys with different types of prepuce retractabilty. A prospective study was performed involving boys referred to our hospital for possible phimosis between January 2004 and February 2008. All were treated initially with fluticasone proprionate 0.05% for a period of 4-8 weeks. Patients were reassessed after 6 months of follow up. A total of 1185 boys with a diagnosis of phimosis were treated with fluticasone proprionate 0.05%. Successful results were achieved in 1079 (91.1%) patients including boys with mild balanitis xerotica obliterans. No side effects were noticed. Our results show that fluticasone proprionate 0.05%, a mild potent corticosteroid, is effective and safe in the treatment of boys with different types of phimosis.

Fluticasone Propionate Protects against Ozone-Induced Airway Inflammation and Modified Immune Cell Activation Markers in Healthy Volunteers

BackgroundOzone exposure induces airway neutrophilia and modifies innate immune monocytic cell-surface phenotypes in healthy individuals. High-dose inhaled corticosteroids can reduce O3-induced airway inflammation, but their effect on innate immune activation is unknown.ObjectivesWe used a human O3 inhalation challenge model to examine the effectiveness of clinically relevant doses of inhaled corticosteroids on airway inflammation and markers of innate immune activation in healthy volunteers.MethodsSeventeen O3-responsive subjects [> 10% increase in the percentage of polymorphonuclear leukocytes (PMNs) in sputum, PMNs per milligram vs. baseline sputum] received placebo, or either a single therapeutic dose (0.5 mg) or a high dose (2 mg) of inhaled fluticasone proprionate (FP) 1 hr before a 3-hr O3 challenge (0.25 ppm) on three separate occasions at least 2 weeks apart. Lung function, exhaled nitric oxide, sputum, and systemic biomarkers were assessed 1–5 hr after the O3 challenge. To determine the effect of FP on cellular function, we assessed sputum cells from seven subjects by flow cytometry for cell-surface marker activation.ResultsFP had no effect on O3-induced lung function decline. Compared with placebo, 0.5 mg and 2 mg FP reduced O3-induced sputum neutrophilia by 18% and 35%, respectively. A similar effect was observed on the airway-specific serum biomarker Clara cell protein 16 (CCP16). Furthermore, FP pretreatment significantly reduced O3-induced modification of CD11b, mCD14, CD64, CD16, HLA-DR, and CD86 on sputum monocytes in a dose-dependent manner.ConclusionsThis study confirmed and extended data demonstrating the protective effect of FP against O3-induced airway inflammation and immune cell activation.

A longitudinal assessment of the effect of inhaled fluticasone propionate therapy on adrenal function and growth in young children with asthma

Fluticasone proprionate (FP) is increasingly used to treat very young children with asthma. Its safety in terms of effects on the hypothalamic pituitary axis (HPA) and growth in this age group is uncertain. Eleven children (median (range) age 10 (5.6-24.3) months) presenting with recurrent wheeze and family history of asthma were studied prospectively for a period of 18 months. Children received daily-inhaled FP 250 microg via a spacer device. No other corticosteroid therapy was administered prior to or during the study. A Short Standard Synacthen Test (SST) (125 microg) was performed pretreatment, and after 6 and 18 months. Weight (Wt), height (Ht), and body mass index (BMI) were measured at 3-6 monthly intervals. Fasting early morning and peak cortisol levels remained within the normal reference range with therapy. There were no changes in Ht SDS, whereas both Wt SDS (baseline 0.05 (-2.17 to 0.52) vs. +18 months 0.68 (-0.5 to 1.36) P < 0.02) and BMI SDS (-0.22 (-1.73 to 0.75) vs. 0.86 (0.03 to 1.99) P < 0.005) increased after 18 months of treatment. Daily treatment with inhaled FP 250 microg in young children with asthma appears to have no adverse effects on the HPA or on linear growth, however, treatment is associated with increases in body Wt and BMI in young children.

Growth recovery after withdrawal of inappropriate inhaled fluticasone proprionate

Pediatric PulmonologyVolume 43, Issue 3 p. 307-308 Growth recovery after withdrawal of inappropriate inhaled fluticasone proprionate Dr. Malcolm Brodlie MB, ChB, Corresponding Author Dr. Malcolm Brodlie MB, ChB [email protected] Department of Respiratory Paediatrics, Freeman Hospital, High Heaton, Newcastle upon Tyne NE7 7DN, United KingdomDepartment of Respiratory Paediatrics, Freeman Hospital, High Heaton, Newcastle upon Tyne, NE7 7DN, United Kingdom.Search for more papers by this authorDavid Spencer MD, David Spencer MD Department of Respiratory Paediatrics, Freeman Hospital, High Heaton, Newcastle upon Tyne NE7 7DN, United KingdomSearch for more papers by this author Dr. Malcolm Brodlie MB, ChB, Corresponding Author Dr. Malcolm Brodlie MB, ChB [email protected] Department of Respiratory Paediatrics, Freeman Hospital, High Heaton, Newcastle upon Tyne NE7 7DN, United KingdomDepartment of Respiratory Paediatrics, Freeman Hospital, High Heaton, Newcastle upon Tyne, NE7 7DN, United Kingdom.Search for more papers by this authorDavid Spencer MD, David Spencer MD Department of Respiratory Paediatrics, Freeman Hospital, High Heaton, Newcastle upon Tyne NE7 7DN, United KingdomSearch for more papers by this author First published: 24 January 2008 https://doi.org/10.1002/ppul.20775Citations: 3AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL No abstract is available for this article.Citing Literature Volume43, Issue3March 2008Pages 307-308 RelatedInformation

Targeting adenosine receptors in the treatment of allergic rhinitis: a randomized, double‐blind, placebo‐controlled study

There is evidence that adenosine plays a role in the pathogenesis of asthma and rhinitis; however, it is currently unclear whether adenosine receptors are useful therapeutic targets in the treatment of allergic airway diseases. The study evaluated the efficacy of intranasal treatment with an adenosine A(2A) receptor agonist/adenosine A(3) receptor antagonist (50 micro g), administered twice daily for 7 days, to reduce nasal symptoms and release of inflammatory mediators following intranasal allergen challenge in patients with allergic rhinitis (AR). The compound was compared with twice-daily treatment with intranasal fluticasone proprionate nasal spray (FPANS) for 7 days. A randomized, double-blind, double-dummy, placebo-controlled, three-way balanced, incomplete block, crossover study was conducted on 48 males with verified AR. Following intranasal challenge with either an extract from the house dust mite (HDM), Dermatophagoides pteronyssinus, rye grass or cat dander, nasal responses and the concentrations of albumin, tryptase, myeloperoxidase, eosinophilic cationic protein, epithelial neutrophil-activating protein-78 (ENA-78), IL-5 and IL-8 in nasal secretions were measured and treatment groups were compared. Drug improved nasal blockage but had no significant effect on rhinorrhoea, number of sneezes or peak nasal inspiratory flow measurements when compared with placebo. Drug reduced tryptase release after EAR but did not significantly reduce the levels of other mediators. A novel agonist/antagonist of adenosine A(2A) and A(3) receptors appears to have limited clinical benefit in both the early-phase and the late-phase response to intranasal allergen challenge. However, reduction of some pro-inflammatory mediators suggests that comparable, more selective compounds may have additional benefits meriting further investigation.

Strategies to screen for adrenal suppression in children with asthma should take account of compliance with inhaled corticosteroids

Since their introduction over 30 years ago inhaled corticosteroids (ICS) have proven to be highly effective in the management of paediatric asthma [2]. Fluticasone proprionate (FP) was introduced in Britain in 1993 as a highly potent ICS. The British National Formulary for children advises a maximum dose of 400 μg/day of FP in children >5 years. However, the British Thoracic Society guidelines in 1997 advised doses of up to 1000 μg/day in children >5 years, and it therefore became common practice amongst British paediatricians to prescribe high doses in difficult cases [4]. The appropriateness of high ‘off-license’ doses has been questioned recently in the aftermath of the publication of a number of cases of life-threatening adrenal suppression (AS) [5]. In the literature the prevalence of AS in children prescribed FP ranges from 12–67% [3]. Compliance with inhaled medication is notoriously poor, and it is possible that the effect of FP may have been underestimated in the past due to a failure to assess compliance. We report the prevalence of AS in children attending a tertiary asthma clinic recommended ≥1000 μg/day FP by their hospital specialist and relate this to compliance in the form of the actual amount prescribed over the last year by their General Practitioner (GP). In the UK hospital-based specialists recommend medications that are subsequently prescribed by GPs who have accurate electronic prescribing records. Adrenal function was assessed by the standarddose (250 μg) Synacthen test. A peak cortisol >530 nmol/l was considered to be an adequate response based on controls from our institution (P.M. Crofton, unpublished observations). Spearman rank correlation was used to explore relations between variables. Twenty-nine children (median age: 11 years; range: 6– 16 years) were recommended by their hospital specialist ≥1000 μg/day FP, three of whom were found to have AS (recommended 2000, 2000 and 1000 μg/day with peak cortisols <50, <50 and 72 nmol/l respectively). Data on the actual prescribed dose compared to the recommended dose are summarised in Table 1. Compliance with medication was 33% in children with and without AS. Overall there was an inverse correlation between actual prescribed daily dose and peak cortisol level (rs−0.44, p=0.03). In two children with AS the FP dose was reduced under hydrocortisone cover with subsequent normalisation of cortisol responses. The third patient moved to adult services, and there is no follow-up data. The finding that 10% of children recommended ≥1000 μg/day had AS is especially relevant in the context of recent life-threatening cases. It is equally worrying but perhaps unsurprising that compliance appears to be so poor at 33%. Although there was a moderate overall inverse Eur J Pediatr (2007) 166:493–494 DOI 10.1007/s00431-006-0260-7

Novel Treatment of Esophageal Crohnʼs Disease with Swallowed Fluticasone Proprionate

Case presentation: A 27 year-old female was referred for evaluation of dysphagia. Symptoms began ten days prior to presentation and occurred predominantly with solids. She denied weight loss, heartburn, abdominal pain, odynophagia, hematemesis or melena. There is no family history of inflammatory bowel disease and she takes no medications. Her physical examination was normal. EGD revealed multiple small ulcers in the middle third of the esophagus. Biopsies revealed nonspecific inflammation with reparative changes. No fungal forms were visualized and immunostains for HSV and CMV were negative. Additional laboratory studies were notable for a platelet count of 521000, ESR of 33 mm/hr and non-reactive HIV antibody. A subsequent small bowel series revealed a nodular polypoid filling defect in the distal terminal ileum and cecum. Colonoscopy revealed a deformed and ulcerated ileocecal valve, congested and edematous cecal mucosa, and multiple superficial ulcerations in the terminal ileum. Biopsies confirmed architectural distortion, cryptitis and crypt abscess formation consistent with Crohn's disease. Given the reported effectiveness of swallowed steroids in eosinophilic esophagitis, the patient was started on a fluticasone propionate inhaler, two sprays swallowed twice daily. The patient was also started on 6-mercaptopurine (6-MP) for her ileocolonic disease. After one week of therapy with fluticasone, her dysphagia had resolved. Repeat EGD showed complete healing of the previously noted esophageal ulcers. Discussion: Crohn's disease involving the esophagus is rare. Most patients also have extra-esophageal involvement. Symptoms may include dysphagia, odynophagia and chest pain. Endoscopic findings are non-specific, but superficial aphthous ulcerations are most common. Strictures and fistulas are seen in severe cases. The most common histologic findings are ulcers and a chronic lymphocytic infiltrate. Optimal management is not well established, but most patients are treated with systemic steroids and antisecretory agents. Immunomodulators have been used in difficult cases. Endoscopic dilation is helpful in stricturing disease, but surgery may be required in refractory cases. There are no previous reports of topical steroids in the management of esophageal Crohn's disease. Although our patient was concomitantly started on 6-MP, the therapeutic effect of this agent is delayed and unlikely to have played a role in the healing of her esophageal ulcers.

Adrenal responses to low dose synthetic ACTH (Synacthen) in children receiving high dose inhaled fluticasone

Background and Aims: Clinical adrenal insufficiency has been reported with doses of inhaled fluticasone proprionate (FP) >400 μg/day, the maximum dose licensed for use in children with asthma. Following two...

Enhancement of Host Defense Gene Expression by Glucocorticoids (GC) is Mediated by CCAAT-Enhancer Binding Protein (C/EBP)

RATIONALE: Previous work showed that GC spare or enhance the expression of molecules associated with host defense and inhibit those associated with inflammation. In this study, we investigated the mechanism of these selective effects.METHODS: BEAS-2B and human primary bronchial epithelial cells were treated with fluticasone proprionate (FP) for 2 hours before stimulation with dsRNA (PolyIC, 25μg/ml) for 18 hours. Responses were monitored by real-time PCR, Western blot, EMSA and luciferase reporter assays. SiRNA was used to reduce the transcription factor C/EBP, which is known to be involved in the hepatic acute phase response.RESULTS: We detected mRNA for C/EBPα,β,γ and δ, but not ε, in resting cells. Both dsRNA and FP selectively induced C/EBPß and C/EBPδ mRNA in BEAS-2B cell and the combination produced a synergistic effect in both primary cells and the BEAS-2B cell line. Western Blot confirmed the induction of C/EBPß by FP and the combination of FP and dsRNA. The maximal effects of FP were observed at 10-9M and after treatment for 4 hours. Luciferase reporter assays and EMSA indicated that FP induced the DNA binding activity and functional activity of C/EBPß. SiRNA targeting C/EBPß and C/EBPδ blocked the enhancement by FP of the expression of the host defense genes CRP, SAA, MBL and C3, but not the inflammatory genes RANTES, GM-CSF, IL-8.CONCLUSIONS: The enhancement of airway host defense responses by GC is mediated in part by C/EBP. RATIONALE: Previous work showed that GC spare or enhance the expression of molecules associated with host defense and inhibit those associated with inflammation. In this study, we investigated the mechanism of these selective effects. METHODS: BEAS-2B and human primary bronchial epithelial cells were treated with fluticasone proprionate (FP) for 2 hours before stimulation with dsRNA (PolyIC, 25μg/ml) for 18 hours. Responses were monitored by real-time PCR, Western blot, EMSA and luciferase reporter assays. SiRNA was used to reduce the transcription factor C/EBP, which is known to be involved in the hepatic acute phase response. RESULTS: We detected mRNA for C/EBPα,β,γ and δ, but not ε, in resting cells. Both dsRNA and FP selectively induced C/EBPß and C/EBPδ mRNA in BEAS-2B cell and the combination produced a synergistic effect in both primary cells and the BEAS-2B cell line. Western Blot confirmed the induction of C/EBPß by FP and the combination of FP and dsRNA. The maximal effects of FP were observed at 10-9M and after treatment for 4 hours. Luciferase reporter assays and EMSA indicated that FP induced the DNA binding activity and functional activity of C/EBPß. SiRNA targeting C/EBPß and C/EBPδ blocked the enhancement by FP of the expression of the host defense genes CRP, SAA, MBL and C3, but not the inflammatory genes RANTES, GM-CSF, IL-8. CONCLUSIONS: The enhancement of airway host defense responses by GC is mediated in part by C/EBP.

Effects of low doses of inhaled fluticasone propionate on inflammation and remodelling in persistent‐mild asthma

In asthma a dysregulation of eosinophil apoptosis and an imbalance of metalloproteinase-9 (MMP-9) and tissue inhibitor metalloproteinase-1 (TIMP-1) play an important role in airway inflammation and remodelling. We evaluated the effects of a low dose of inhaled fluticasone proprionate (FP) (100 microg bid by Diskus) for 4 weeks in 24 steroid naive patients with mild persistent asthma, symptomatic and with a sputum eosinophilia >or=3% on clinical outcomes and inflammatory markers such as the induced sputum eosinophils, the induced sputum apoptotic eosinophils, the levels of MMP-9 and TIMP-1 and their molar ratio in the induced sputum supernatants. After FP treatment forced expiratory volume (FEV1) and FEV1/forced vital capacity values, PEF (L/min), sputum apoptotic eosinophils, and MMP-9/TIMP-1 molar ratio in sputum supernatants of asthmatic subjects were significantly increased in comparison with baseline, while sputum eosinophils significantly decreased. Change (Delta) in FEV1 after treatment with FP negatively correlated with the Delta in sputum eosinophils, while the Delta in MMP-9 values positively correlated with Delta in TIMP-1 values. This study shows that the clinical improvement achieved by the use of low doses of FP in asthmatics is related, at least in part, to the resolution of eosinophilic inflammation and the downregulation of remodelling markers.

Interleukin-10–Secreting “Regulatory” T Cells Induced by Glucocorticoids and β2-Agonists

Greater clinical benefit in controlling the symptoms of asthma is frequently observed through combining moderate doses of inhaled glucocorticoids together with long-acting beta(2)-agonists, as compared with increasing glucocorticoid dosage alone. To address in vitro whether glucocorticoids plus beta(2)-agonists, compared with glucocorticoids alone, have greater inhibitory activity on CD4+ T cell responses to allergen, peripheral blood CD4+ T cell responses to allergen were compared in the presence or absence of the glucocorticoid fluticasone proprionate and the short- and long-acting beta(2)-agonists salbutamol and salmeterol, respectively. Fluticasone proprionate inhibited interleukin (IL)-5 and IL-13 and enhanced IL-10 synthesis in allergen-stimulated cultures in a concentration-dependent manner. Salmeterol, but not salbutamol, inhibited IL-5 and IL-13 and enhanced IL-10 synthesis in these cultures. When used in combination the two drugs demonstrated an additive effect on this pattern of cytokine production. Allergen-specific T cell lines induced in the presence of salmeterol and fluticasone proprionate inhibited IL-5 and IL-13 production by allergen-specific Th2 cell lines in an IL-10-dependent manner. Thus fluticasone proprionate and salmeterol increased IL-10 and reduced Th2 cytokine synthesis additively in allergen stimulated human CD4+ T cells.

Dilated cardiomyopathy with mural thrombus presenting with asthma-like symptoms

Rationale We report a case of dilated cardiomyopathy with mural thrombus presenting in an atopic patient with symptoms of reactive airways disease. Methods A 47 year-old female presented to the Allergy Clinic during the spring season with rhinitis, ocular pruritus, cough, chest tightness and wheeze. Allergy skin testing showed reactivity to tree pollens and dust mites. A chest film showed a mild lingular infiltrate. Pulmonary function test showed mild obstructive airway disease. She was treated with azithromycin, beclomethasone diproprionate 80 ug, 2 puffs twice daily, albuterol as needed and a short Prednisone taper. Her symptoms improved. Four weeks later, she experienced a recurrence of cough, nocturnal wheeze, dyspnea and tachycardia. When her symptoms persisted despite the use of fluticasone proprionate,220 ug, salmeterol and monteleukast, a repeat chest film was obtained. Results The chest film showed a mild cardiomegaly compared with her previous chest x-ray. An echocardiogram revealed a dilated cardiomyopathy with a left ventricular mural thrombus. D-Dimer was positive. Doppler study of the lower extremities was normal. She was started on digoxin, furosemide, warfarin sodium and lisinopril and has done well off all asthma medications. Conclusions Allergists should maintain a high index of suspicion in cases where symptoms suggestive of reactive airway disease may be masquerading as another unexpected disease entity.

Tissue Remodeling and Corticosteroids: Fluticasone Proprionate Inhibits Nicotine-Induced Fibronectin Expression in Fibroblasts

Tissue Remodeling and Corticosteroids: Fluticasone Proprionate Inhibits Nicotine-Induced Fibronectin Expression in Fibroblasts

Tissue Remodeling and Corticosteroids: Fluticasone Proprionate Inhibits Nicotine-Induced Fibronectin Expression in Fibroblasts

Tissue Remodeling and Corticosteroids: Fluticasone Proprionate Inhibits Nicotine-Induced Fibronectin Expression in Fibroblasts

Body composition and growth in asthmatic children treated with inhaled steroids

Prolonged treatment with inhaled steroids is recommended for long-term control of asthma in children; however, it can interfere with growth and body composition. The aim of this study is to answer the question whether 6 months treatment with inhaled steroids causes body fat accumulation and growth velocity reduction. Hospital-based, open study of body composition [by dual-energy X-ray absorptiometry (DXA), bioelectrical impedance analysis (BIA) and skinfolds] and growth of 26 asthmatic children, treated for 6 months with inhaled steroids [budesonide (BUD) 400 microg/day (group 1) or fluticasone proprionate (FP) 200 microg/day (group 2)], sodium cromoglycate and beta2-agonist (salbutamol) compared with a control group of 16 asthmatic children treated only with sodium cromoglycate and beta2-agonist. On average, total and regional fat mass, adjusted for pubertal stage and gender, and growth velocity were similar in all three groups of patients and were not influenced by treatment (% mean change +/- 1 SD of fat mass during treatment in BUD 0.1 +/- 3.0%, FP -1.1 +/- 3%, and control -2.8 +/- 3.5%; ANOVA P > or = .05); however seven patients, two in group 1 (1 preschool child), three in group 2 (2 preschool children) and two in the control group (two prepubertal boys aged 8.5 and 9.5 year), during treatment, showed a growth velocity standard deviation score below the third percentile. A 6-month treatment with inhaled BUD and FP does not induce body fat accumulation; however, in a few preschool children the treatment was associated with growth velocity below the third percentile. Our results suggest the need for constant monitoring of growth in all asthmatic children on chronic treatment with inhaled steroids. Further studies devoted to the effects of inhaled steroids use in preschool children are needed.

614 The effect of fluticasone proprionate nasal spray as adjunctive therapy to cefuroxime axetil on the resolution of radiographic evidence of an acute episode of sinusitis

614 The effect of fluticasone proprionate nasal spray as adjunctive therapy to cefuroxime axetil on the resolution of radiographic evidence of an acute episode of sinusitis

Glucocorticoids ablate IL-1beta-induced beta-adrenergic hyporesponsiveness in human airway smooth muscle cells.

We have previously reported that interleukin (IL)-1beta decreases responsiveness of cultured human airway smooth muscle (HASM) cells to beta-agonists. The purpose of this study was to determine whether glucocorticoids inhibit this IL-1beta effect. Dexamethasone (Dex; 10(-6) M) had no effect on concentration-related decreases in cell stiffness in response to isoproterenol (Iso) in control cells as measured by magnetic twisting cytometry but prevented the decreased responsiveness to Iso observed in IL-1beta (20 ng/ml)-treated cells. In addition, Dex had no effect on Iso-stimulated cAMP formation in control cells but prevented the IL-1beta-induced reduction in Iso-stimulated cAMP formation. Similar effects on cell stiffness and cAMP responses were seen after pretreatment with the glucocorticoid fluticasone proprionate (FP). Dex and FP also prevented IL-1beta-induced hyporesponsiveness to PGE(2) stimulation. In contrast, neither IL-1beta nor glucocorticoids had any effect on cell stiffness responses to dibutyryl cAMP. We have previously reported that the IL-1beta effect on beta-adrenergic responsiveness is mediated through cyclooxygenase-2 expression and prostanoid formation. Consistent with these observations, IL-1beta-induced cyclooxygenase-2 expression was virtually abolished by FP at concentrations of 10(-10) M and greater, with a resultant decrease in PGE(2) formation. However, Dex did not inhibit IL-1beta-induced nuclear translocation of nuclear factor-kappaB or activator protein-1 in HASM cells. In summary, our results indicate that, in HASM cells, glucocorticoids alone do not alter responses to beta-agonists but do inhibit IL-1beta-induced beta-adrenergic hyporesponsiveness. Glucocorticoids mediate this effect by inhibiting prostanoid formation but without altering nuclear factor-kappaB or activator protein-1 translocation.

Ligand-induced differentiation of glucocorticoid receptor (GR) trans-repression and transactivation.

Inhaled glucocorticosteroids (GCs) are the most effective therapy for the control of asthma and are now advocated as first line therapy for most patients. Steroids reduce airway hyper-responsiveness and suppress the airway inflammatory response in asthma, but their exact mechanisms and cellular targets in the lung are uncertain 111. Classically GCs bind to a cytosolic 94kDa receptor protein, that is held in an inactive, non-DNA binding state by a 9OkDa heat shock protein (hsp90). With lioand binding, the DNA binding site is revealed and enhancement of the transcriptional response occurs following the translocation of active receptor to the nucleus. The GR binds to a consensus cis-acting DNA sequence AGAACAnnnTGTTCT (GRE) in the 5’ upstream controlling region of the genes under positive control by GCs causing an increase in gene transcription [2, 31. The mechanisms involved in gene repression are less well understood. In some instances, this may be because GR binding to a negative GRE results in steric hindrance to the binding of transcription factors that may induce the same gene. GR may also form complexes with activating transcription factors in the nucleus to inhibit their effect and so modulate genes which contain no GREs. Pro-inflammatory transcription factors, such as AP-1 and NF-KB, can form complexes with activated GR. causing mutual repression of DNA binding or affects interaction with the basal transcription apparatus thereby reducing steroid responsiveness [4-6]. Furthermore, GCs can upregulate the synthesis of I-KB, the cytosolic inhibitor of NF-KB [7]. Conversely, GCs may inhibit the effects of those cytokines that produce their effects on gene transcription via activation of AP-1 or NF-KB. For example, in human lung and in peripheral blood monocytes, TNFa and phorbol esters increase AP-1 and NF-KB binding to DNA and this is inhibited by dexamethasone [6, 81. Cytokines play an important role in chronic inflammation and the pattern of cytokine expression largely determines the nature and persistence of the inflammatory response. GCs have potent inhibitory effects on cytokine mediated inflammation and inhibit the transcription of several cytokines that are relevant in chronic inflammation, including interleukin-1 (IL-l), TNFa, GM-CSF, 11-3, 11-4, IL-5, 11-6 and IL-8 [3]. Other members of the steroid superfamily also act in an analogous manner and, in the case of retinoids, the ligand is able to modulate the ability of the receptor to bind to DNA and to AP-1 [9]. We have, therefore, investigated the ability of two glucocorticoids, dexamethasone and fluticasone, to influence GR-induced inhibition of NF-KB and AP-1 DNA binding and on the release of GM-CSF, an NF-KB-inducible gene in A549 cells. GM-CSF release was measured following stimulation with lng/ml IL-lp. Furthermore, we examined the ability of GR to bind to its DNA binding motif. Lung epithelium-like A549 cells were cultured in Dulbecco’s Modified Eagles medium (DMEM) in the presence of 2% foetal calf serum (FCS) until 70-80% confluent. FCS was removed from the culture medium and cells cultured for a further 2 days. Equal numbers of A549 cells were incubated with a range of concentrations of dexamethasone (Dex) and fluticasone propionate (FP) for 1-24 hours in the presence or absence of lnglml of IL-lp. Control experiments using cells incubated in serum free media alone were run in parallel. At every time point cells were collected by microcentrifugation and nuclear protein extracted [6]. Double stranded oligonucleotides encoding the consensus target sequence for GRE, AP-1 and NF-KB were end-labelled using t2P]-ATP and T4 polynucleotide kinase. l pg of nuclear protein from each sample was incubated with 50,000 cpm of labelled oligonucleotide. Protein-DNA complexes were separated on 6% polyacrylamide gels. The retarded band was detected by autoradiogaphy and quantified by laser densitometry. All band density measurements were then expressed as a percentage of initial binding. Specificity was determined by addition of excess unlabelled double stranded oligonucleotides. GM-CSF concentrations in culture supernatants were measured using a specific GM-CSF ELISA calibrated with human recombinant GM-CSF (0-2000pg/ml). Sensitivity of the assay was 8pglml. FP and Dex both caused a time and dose dependent inhibition of IL1p-stimulated GM-CSF release. Inhibition was first detected at 4 hrs post-steroid treatment and was at maximal levels at all time points from 10-24 hrs. At lOhrs FP caused a dose-dependent inhibition of IL-1pstimulated GM-CSF release with complete inhibition at lO-”M (EC= 5.7il.8 x lo’”), whilst Dex inhibition of IL-1p-stimulated GM-CSF release was found to be maximal at 10aM (EC= 1.2iO.5 x 10.9. This showed a 21 1-fold shift in responsiveness in spite of there being only an Figure 1. The effects of dexamethasone (Dex) and Fluticasone Proprionate (FP) on GMCSF release into the Supernatant of A549 cells.

Mast cells and mediators in the nasal mucosa after allergen challenge. Effects of four weeks' treatment with topical glucocorticoid

The study focuses on the relationship between the tissue density of mast cells, the tissue histamine levels and the levels of markers of mast cell activation after an allergen challenge of the nasal mucosa of allergic patients. The effect of 4 weeks' treatment with a topical glucocorticoid, fluticasone propionate, was studied in a double-blind, placebo-controlled study of 25 hay fever patients. Nasal biopsies were obtained before and after the treatment period for the evaluation of mast cell density and tissue histamine levels. Nasal challenges were performed at 2-week intervals for 8 weeks using a standardized nasal lavage model. TAME-esterase was analysed in the returned lavage fluid from all the challenges (weeks 0-8), while the levels of histamine and tryptase were analysed in lavage fluids from challenges performed before and after the treatment period (weeks 0 and 4). The symptoms of nasal allergy were assessed after each challenge. Treatment with fluticasone propionate did not influence mast cell density, the tissue histamine concentration, the lavage histamine levels or the TAME-esterase activity, while a reduction in nasal symptoms and tryptase in nasal lavage fluid was revealed. Our present study again emphasizes the fact that the mast cell is an important trigger cell in the immediate nasal allergic response. The study also demonstrates the usefulness of the measurements of tryptase as an indicator of both mast cell activation and the efficacy of topical steroid treatment.