Eosinophil Cationic Protein Concentrations Research Articles

Airway Surfactant Protein D Deficiency inAdults With Severe Asthma.

Surfactant protein D (SP-D) is an essential component of the innate immune defense against pathogens within the airways. SP-D also regulates allergic inflammation and promotes the removal of apoptotic cells. SP-D dysregulation is evident in several pulmonary diseases. Our aim was to investigate whether airway and serum levels of SP-D are altered in treatment-resistant severe asthma. SP-D concentrations were measured in matched serum and BAL samples collected from 10 healthy control subjects (HC) and 50 patients with asthma (22 with mild asthma [MA] and 28 with severe asthma [SA]). These samples were also evaluated by using Western blot analysis to investigate variations in SP-D size. SP-D levels in BAL samples were significantly lower in SA compared with HC and MA (P< .001) and inversely correlated with BAL eosinophil cationic protein concentrations in SA (P< .01). Serum SP-D was significantly increased in SA compared with HC and MA (P< .001), and BAL/serum ratios were significantly lower in SA compared with HC and MA(P< .001). Reduced SP-D levels in BAL samples, with concomitant increases in serum in SA, were associated with degraded fragments of SP-D in the serum and increased BAL neutrophil counts and lipopolysaccharide levels. These findings suggest defective innate immunity within the airways in SA, as reflected by low BAL SP-D concentrations and altered bacterial presence with airway neutrophilia. Furthermore, BAL SP-D leakage into the serum in patients with SA may provide a peripheral blood biomarker, reflecting increased epithelial damage and/or epithelial permeability within the peripheral airways.

Reductions in eosinophil biomarkers by benralizumab in patients with asthma

BackgroundEosinophilic inflammation is frequently associated with increased asthma severity. Benralizumab is a humanized, afucosylated, anti–interleukin-5Rα monoclonal antibody that selectively depletes eosinophils and basophils through enhanced antibody-dependent cell-mediated cytotoxicity. ObjectiveTo study effects of benralizumab on eosinophil counts and activity following administration to asthma patients. MethodsSera were collected from asthma patients enrolled in two clinical studies. Placebo or benralizumab was subcutaneously administered to patients in Phase I (100 or 200 mg, multiple doses; N = 14; NCT00659659) and Phase IIa (25, 100, or 200 mg every 4 weeks; N = 24; NCT00783289) studies. Sera were also collected from healthy volunteers (N = 20) for comparison. Blood eosinophils, IL-5, eosinophil-derived neurotoxin (EDN), eosinophil cationic protein (ECP), eotaxin/chemokine (C–C motif) 11 (CCL11), eotaxin-2/CCL24, tumor necrosis factor (TNF), and interferon-γ (IFN-γ) were measured at baseline and post-treatment. ResultsIncreased EDN concentrations were observed in sera of patients from both studies relative to healthy volunteers (p < 0.05). At baseline, sera EDN concentrations correlated with blood eosinophil counts (rs = 0.5; p < 0.05). Benralizumab reduced blood eosinophil numbers and sera EDN and ECP relative to baseline (p < 0.05). No changes in TNF or IFN-γ were observed, while serum IL-5, eotaxin/CCL11, and eotaxin-2/CCL24 increased after benralizumab administration vs. placebo (p < 0.05). ConclusionsIn two independent studies, serum IL-5, EDN, and ECP were modulated following benralizumab. Eosinophil depletion after benralizumab also resulted in significant reductions in EDN and ECP concentrations, suggesting that cytotoxic granule proteins were not released after eosinophil reduction.

Comorbidity has no impact on eosinophil inflammation in the upper airways or on severity of the sinonasal disease in patients with nasal polyps.

The study was designed to determine whether there is an association between the comorbidity as atopy, bronchial asthma, aspirin intolerance and eosinophil infiltration of the upper airways, severity of the sinonasal disease and rate of revision sinus surgery in patients with nasal polyps. One hundred and fifty patients were enrolled in the prospective study. Differences in CT score, rate of revision surgery, concentration of eotaxin and eosinophil cationic protein in nasal lavage fluid (NALF) and distribution of eosinophils in NALF and nasal tissue in patients with chronic rhinosinusitis with nasal polyps (CRSwNP), chronic rhinosinusitis without nasal polyps (CRSsNP) and control group were investigated. We focused on the relationship between presence of comorbidity (atopy, bronchial asthma and aspirin intolerance) and severity of the disease, the need of revision surgery and markers of eosinophil inflammation in upper airways in patients with CRSwNP. Patients with CRSwNP had more severe form of the sinonasal disease, higher rate of revision FESS and significant higher presence of markers of eosinophil inflammation in NALF and nasal tissue than patients with CRSsNP (P<0.05). Atopic and non-atopic asthma as wellas aspirin sensitivity significantly more often coexisted with CRSwNP. Comorbidity did not influence eosinophil infiltration or severity of the disease in patients with CRSwNP. Presence of comorbidity (atopy, bronchial asthma and aspirin intolerance) has no impact on severity of the disease or eosinophil content in the upper airways in patients with CRSwNP.

Virus detection and cytokine profile in relation to age among acute exacerbations of childhood asthma

BackgroundLittle information is available regarding eosinophil activation and cytokine profiles in relation to age in virus-induced bronchial asthma. We therefore explored the association between age, respiratory viruses, serum eosinophil cationic protein (ECP), and cytokines/chemokines in acute exacerbations of childhood asthma. MethodsWe investigated viruses in nasal secretions from 88 patients with acute exacerbation of childhood asthma by using antigen detection kits and/or RT-PCR, followed by direct DNA sequencing analysis. We also measured peripheral eosinophil counts, and the serum levels of ECP and 27 types of cytokines/chemokines in 71 virus-induced acute asthma cases and 13 controls. ResultsViruses were detected in 71(80.7%) of the 88 samples. The three major viruses detected were rhinoviruses, RS viruses, and enteroviruses; enteroviruses were found to be dominant in patients aged ≥3 years. There was no change in the levels of rhinoviruses and RS viruses between the two age groups, defined as children aged <3 years and children aged ≥3 years. Serum concentrations of ECP, IL-5, and IP-10 were significantly elevated in virus-induced acute asthma cases compared with controls. Serum ECP values were significantly higher in patients with virus-induced asthma at age ≥3 years compared with those aged <3 years. Among the 27 cytokines/chemokines, serum IP-10 was significantly higher in virus-induced asthma in patients <3 years than in those ≥3 years. Serum ECP and IL-5 production correlated significantly with age, whereas serum IP-10 showed an inverse correlation with age. ConclusionsAge-related differences in cytokine profiles and eosinophil activation may be related to virus-induced acute exacerbations of childhood asthma.

Role of Synergism between RV Infection and Atopy in the Pathophysiology of Intermittent Asthmatic Bronchitis in Adolescent Group in Mansoura

Airway inflammation represents the corner stone in the pathogenesis of asthma which reflects the different aspects of the disease. Several studies had attributed several aspects in asthma pathogenesis to the interplay between Th1 and Th2 immune responses. Cytokines released by Th2 cells seem to play very important role in inducing allergic air way inflammation. To test the hypothesis that RV infection is responsible for asthma exacerbation in adolescent group and to test the relationship between RV infection and allergen sensitization. Seventy five adolescents were divided into 5 groups: control healthy group, group with controlled asthma without viral infection, viral infected group without asthma, asthmatic group with viral infection without atopy, and asthmatic group with viral infection with atopy. For all groups sputum samples were analyzed for eosinophile percentage, eosinophilic cationic protein concentration, IL 13 and IFy concentration using ELISA and RVRNA assay using real time PCR. Serum total IgE levels were also detected. Using the spirometer forced expiratory volume in one second (FEV1)/forced vital capacity (FVC) % was measured for all groups. Asthmatic group with viral infection and atopy showed a heavy viral load in the sputum together with severe respiratory tract symptoms and asthma symptoms requiring hospitalization. This group also showed higher median serum total IgE levels as compared with other groups (P<0.05), significant higher IL13, eosinophil, and eosinophilic cationic protein concentration in the sputum (P<0.05) and significant lower IF-y concentration in the sputum (P<0.05) as compared with other groups but not significant lower IF-y concentration as compared with asthmatic group with viral infection without atopy. Asthmatic exacerbation in adolescent group with intermittent asthma is attributed to RV infection. RV and allergy act synergistically to worsen both viral respiratory tract symptoms and asthma symptoms. In this situation higher IL13 and total IgE levels together with lower IF-y level play an important role.

Indoor Air Pollutant Exposure and Eosinophil Cationic Protein as an Upper Airway Inflammatory Biomarker among Preschool Children

The upper and lower airways of the respiratory tract are functionally linked, with inflammation in the former playing a vital role in the pathogenesis of asthma and allergy. Studying the association between indoor air pollutants with upper airway inflammation in children will help improve childhood asthma and allergy management related to poor indoor air quality. A cross-sectional study was conducted among preschool children in industrial (Kelana Jaya and Shah Alam) and suburban (Semenyih and Hulu Langat) areas in Selangor, Malaysia. A questionnaire adapted from the American Thoracic Society and International Study on Asthma and Allergy in Children was distributed to obtain the respondents’ background information, school, and home environment. Eosinophil cationic protein (ECP) concentrations in nasal swab samples were collected and analyzed to determine the prevalence of upper airway inflammation. An indoor air quality (IAQ) assessment was also conducted in seven preschools in both industrial and suburban areas, including parameters such as particulate matter up to 10 μm in size (PM10), volatile organic compounds (VOCs), total mold, total bacteria, relative humidity, and air temperature. Statistical analysis shows significant differences in PM10, total mold, total bacteria and relative humidity between the study areas (p < 0.05). The ECP levels among respondents vary significantly between study areas (t = 8.473, p < 0.001). The VOC concentration and ECP level are significantly correlated(prevalence ratio 6.41; 95% CI, 1.268 to 32.394) after controlling all confounders. This study concludes that exposure to indoor air pollutants increases the risk of respiratory problems and may have an impact on the inflammatory and secretory response of the nasal mucosa.

Effect of eosinophil cationic protein on human oral squamous carcinoma cell viability.

The exact function of eosinophils in cancer, particularly in oral squamous cell carcinoma (OSCC), has not yet been elucidated and the possible antitumor effect of these leukocytes is associated with the release of cytotoxic proteins, particularly eosinophil cationic protein (ECP). The aim of this study was to evaluate the effect of ECP on human OSCC lines and to provide novel insights into the role of eosinophils in these tumors. The viability of the SCC-4 and SCC-25 OSCC cell lines was assessed by colorimetric assay using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). The cells were plated into 96-well plates in Dulbecco's modified Eagle's medium/Ham's nutrient mixture F12 supplemented with 10% fetal bovine serum. After 24 h, the indicated concentration of ECP (0-10 µ M) was added to each sample. The plate was read using a microplate reader at a wavelength of 570 nm. The association between variables was estimated by linear regression analysis. There was a significant inverse association between ECP concentrations with SCC-4 (β=0.16, P=0.019) and SCC-25 cell viability (β=0.24, P=0.006). To the best of our knowledge, the present study was the first to investigate the effects of ECP on OSCCs and to demonstrate a significant inverse association between ECP concentrations with SCC-4 and SCC-25 cell viability.

Controlled trial for long-term low-dose erythromycin after sinus surgery for chronic rhinosinusitis.

The efficacy of macrolides in chronic rhinosinusitis (CRS) is still under controversy. To date, only two double-blind, placebo-controlled studies have been published with differing results. None of these studies investigated the possible benefit of macrolides in the postoperative period. We conducted an investigator-initiated clinical trial using 250-mg erythromycin once a day over a period of 3 months, beginning the administration of either erythromycin or placebo 2 weeks after a surgical intervention for CRS. Randomized double-blind, placebo-controlled trial. The concentrations of eosinophilic cationic protein (ECP) and myeloperoxidase in nasal secretion were chosen as primary outcome measures. Additionally, as a secondary outcome measure, changes in the Sino-Nasal Outcome Test-20 score, olfaction, saccharin transit time, nasal endoscopy score, and self-rating of nasal health using a visual analogue scale were evaluated. Sixty-seven patients after surgery for CRS with or without nasal polyps were screened, and 58 patients were randomized to the study groups. For the primary outcomes, the concentrations of ECP changed from 176.4 µl/l ± 79.0 to 226.1 µl/l ± 200.6 in the erythromycin group and from 186.9 µl/l ± 36.0 to 192.9 µl/l ± 189.2 in the placebo group; no statistical differences were found. Of the secondary outcomes, only the nasal endoscopy score showed a statistically significant improvement in the erythromycin group (from 2.6 ± 1.4 to 1.9 ± 1.5 points) compared to the placebo group (from 2.5 ± 1.3 to 2.6 ± 1.5 points). The subgroup of patients without nasal polyps in the erythromycin group showed a tendency to improvement in some secondary outcome criteria. A general recommendation for long-term, low-dose erythromycin treatment after surgery for CRS cannot be given. In patients with CRS without nasal polyps, a tendency to improved parameters was detected. Ib.

Effects of theophylline plus salmeterol/fluticasone propionate combination therapy on lung function and airway inflammation in patients with asthma

Objective To explore the effects of theophylline plus salmeterol/fluticasone propionate combination product (SFC) on clinical control, lung function and airway inflammation in asthmatics. Methods A total of 146 asthmatics received 200 mg theophylline plus 50/250 μg SFC twice daily for 24 weeks. The level of asthma control was assessed by the asthma control test. Testing of lung function and inflammatory markers in induced sputum were performed. And 142 asthmatics received 50/250 μg SFC twice daily for 24 weeks as control. Results Asthma was completely controlled in 61 and 59 in the theophylline plus SFC and SFC groups respectively after a 24-week treatment period (P>0.05). Theophylline plus SFC improved the FEF25%-75% value, indicating small airway function, to a greater extent than SFC [(66.7±18.2)% & (56.6±17.4)%, P<0.01]. Percentage of eosinophil and concentration of eosinophil cationic protein in induced sputum were significantly lower in the theophylline plus SFC group than those in the SFC group [(4.1±2.3)% vs. (6.2±2.7)% & (63.9±39.4) vs. (90.3±46.2) μg/L respectively] (all P<0.01). Conclusion The therapy of theophylline plus SFC may provide greater improvements in small airway function and airway inflammation. Key words: Asthma; Theophylline; Drug therapy, combination

Development of a suspension array assay in multiplex for the simultaneous measurement of serum levels of four eosinophil granule proteins

The concentrations of major basic protein (MBP), eosinophil cationic protein (ECP), eosinophil derived neurotoxin (EDN) and eosinophil peroxidase (EPO) have been associated with eosinophilic disease severity. Whereas a variety of techniques have been used to measure individual eosinophil granule protein concentration, none of these methods efficiently measures MBP, ECP, EDN and EPO simultaneously. A multiplex suspension array system was developed to simultaneously measure the concentrations of MBP, ECP, EDN and EPO in serum. The assay showed excellent inter- and intra-assay reliability, and serum levels of MBP, ECP and EDN from eosinophilic subjects analyzed by ELISA and multiplex were highly correlated (r=0.8579; P<0.0001, r=0.6356; P=0.0006 and r=0.8600; P<0.0001, respectively, Spearman rank correlation). Moreover, the multiplex assay required 500-fold less serum than a single ELISA to achieve comparable sensitivity. Absolute eosinophil count and eosinophil surface expression of the activation marker, CD69, were significantly correlated with concentrations of MBP, EDN and EPO, but not ECP, in serum from eosinophilic subjects. Furthermore, subjects with eosinophilic gastrointestinal disorder and normal peripheral absolute eosinophil counts (<0.5×109/l) had significantly increased concentrations of MBP (P<0.0001), ECP (P<0.0001), EDN (P=0.0001) and EPO (P<0.0001) compared to normal donors. In summary, the eosinophil granule protein multiplex assay provides a rapid and reliable way to measure eosinophil granule protein levels and should prove useful in assessing patterns of degranulation in patients with eosinophilic disorders.

Efficacy of long‐term low‐dose macrolide therapy in preventing early recurrence of nasal polyps after endoscopic sinus surgery

This study assessed efficacy of clarithromycin "long-term" macrolide therapy as an adjunct to maintenance therapy with nasal corticosteroids to prevent recurrence of nasal polyps (NP) after functional endoscopic sinus surgery (FESS). A total of 66 patients with chronic rhinosinusitis and bilateral NP were randomized into 3 study arms, 22 patients in each arm. After FESS, patients in the first and second groups were treated with clarithromycin 250 mg/day for 12 and 24 weeks, respectively, whereas patients in the third group did not receive any clarithromycin. Patients in all 3 groups received maintenance therapy with mometasone furoate 400 μg/day. Patient assessment was conducted before the surgery and 6, 12, and 24 weeks after surgery, using a visual analogue scale (VAS), 20-item SinoNasal Outcome Test (SNOT-20), acoustic rhinometry, rhinomanometry, saccharin transit time, nasal endoscopy, computed tomography (CT) of paranasal sinuses, and measurement of the level of eosinophil cationic protein (ECP) in their nasal secretions. The study confirmed efficacy of "long-term" macrolide therapy, resulting in significant improvement of all parameters except acoustic rhinometry and VAS in both clarithromycin groups as compared to the control. Concentration of ECP in the nasal secretions increased dramatically after surgery, then returned to baseline levels after 12 and 24 weeks of treatment with clarithromycin. In the control group, ECP level continued to increase and was significantly higher at the endpoint. Both groups with clarithromycin showed significantly better endoscopic and CT scores than the control group at the end point. "Long-term" low-dose clarithromycin 250 mg/day is able to control eosinophilic inflammation and prevent early relapse of NP after FESS.

Eosinophil cationic protein levels parallel coagulation activation in the blister fluid of patients with bullous pemphigoid

Bullous pemphigoid (BP) is an autoimmune blistering disease due to autoantibodies against two hemidesmosomal antigens, namely BP180 and BP230, and characterized by coagulation activation both at cutaneous and systemic levels. Skin-infiltrating eosinophils contribute to bulla formation and, upon activation, are supposed to initiate the coagulation cascade. The aim of this study was to investigate whether the activation of eosinophils and coagulation are linked in BP. We evaluated the correlation between eosinophil cationic protein (ECP) levels and concentrations of the prothrombotic markers F1 + 2 and D-dimer in blister fluid and blood samples of 30 BP patients. Thirty healthy subjects were used as normal controls. ECP, F1 + 2 and D-dimer plasma levels were significantly higher in BP patients than in normal subjects. A significant correlation was found between ECP plasma levels and blood eosinophil count (r = 0.54, P = 0.002). F1 + 2 plasma levels positively correlated with disease severity, expressed as the percentage of body surface area involved (r = 0.36, P = 0.048). A striking increase in ECP (288.8 ± 45.2 ng/mL), F1 + 2 (31 409.9 ± 2929.4 pmol/L) and D-dimer levels (342 798.3 ± 44 206 ng/mL) was found in blister fluid from BP patients. In blister fluid, ECP levels were significantly higher than in peripheral blood (P < 0.0001) and were positively correlated with the levels of both F1 + 2 (r = 0.4, P = 0.02) and D-dimer (r = 0.5, P = 0.0045). ECP levels are strikingly elevated in blister fluids from BP patients and correlate with markers of coagulation activation, supporting the view that eosinophils initiate the coagulation cascade at skin level.

Effect of omalizumab on biomarkers in middle ear effusion in patients with eosinophilic otitis media

Conclusions: Eosinophil cationic protein (ECP) concentrations in middle ear effusion (MEE) in patients with eosinophilic otitis media (EOM) were significantly decreased at 3 months after the administration of omalizumab from the baseline level (p < 0.05). This study provides new evidence that omalizumab reduces eosinophilic inflammation in the middle ear and that the reduction of ECP may not be caused by suppression of interleukin (IL)-5 production in the middle ear mucosa. Objective: EOM is an intractable otitis media characterized by a highly viscous effusion containing eosinophils. We recently reported that anti-IgE therapy using omalizumab was efficacious in the treatment of EOM. To clarify the underlying mechanism, we determined changes in biomarkers in MEE related to eosinophilic inflammation after therapy. Methods: Nine patients with EOM received the anti-IgE agent omalizumab for 3 months. Among them, five patients continued anti-IgE therapy for longer than 1 year. Eight EOM patients without administration of omalizumab were also included in the study as controls. The concentrations of eosinophilic inflammatory markers such as ECP, IgE, IL-4, and IL-5 in MEE were measured before and after the administration of omalizumab. Results: After 3 months of omalizumab therapy, the ECP concentration in MEE was significantly reduced from the baseline level (p < 0.05), while no significant change of ECP in the serum was observed. The concentrations of IL-4 and IL-5 in MEE showed no significant change before and after the therapy in EOM patients treated with omalizumab.

The Effects of Phototherapy on Eosinophil and Eosinophilic Cationic Protein in Newborns With Hyperbilirubinemia

Newborns with jaundice requiring or not requiring phototherapy (PT) are at greater risk of developing asthma later in life. In this study, we investigated the effect of PT treatment on eosinophil and eosinophilic cationic protein (ECP) levels in newborns with severe hyperbilirubinemia. Thirty newborns diagnosed with severe hyperbilirubinemia and exposed to light-emitting diode (LED) PT were enrolled into the study. Total serum bilirubin (TSB) levels, complete blood count and serum ECP concentrations were measured before and after PT. TSB and hemoglobin (Hb) counts were lower after PT (p = 0.001). There was no difference between leukocyte, lymphocyte, neutrophil and platelet count before and after PT. Eosinophil levels were increased after PT, although not significantly. ECP levels were higher after PT (p = 0.006). It may be speculated that newborns treated with LED PT, increased ECP might play a role in developing allergic diseases later in life.

The production of the eosinophil proteins ECP and EPX/EDN are regulated in a reciprocal manner.

Previous studies showed that the biological activity and the eosinophil content of eosinophil cationic protein (ECP, RNase 3) are determined by single-nucleotide polymorphisms (SNPs) in the ECP (RNase3) gene. In this study, we report the prevalence of a common SNP in the eosinophil protein x/eosinophil-derived neurotoxin (EPX/EDN, RNase2) and the association with the cellular contents of EPX/EDN and ECP. The genes were sequenced and the EPX/EDN405(G>C) rs2013109 SNPs were also determined by TaqMan 5′nuclease allelic discrimination assay. ECP and EPX/EDN in purified eosinophils or in whole blood extracts were analysed by sensitive immunoassays. The study included 379 non-allergic and allergic subjects. The genotype prevalence of the EPX/EDN405(G>C) polymorphism was GG 59%, GC 36% and CC 6%. The cellular contents of ECP and EPX/EDN were related in a reciprocal fashion with the sums of the protein contents being constant. The contents were associated with the ECP562(G>C) rs2233860 and EPX/EDN405(G>C) gene polymorphisms. The cellular content of eosinophil peroxidase (EPO) was not associated with the ECP and EPX/EDN genotypes. The prevalence of the EPX/EDN405(G>C) genotypes and the contents of the proteins were similar in non-allergic and allergic subjects.The production and storage of the two ancestral proteins, ECP and EPX/EDN likely share common regulatory mechanisms, which result in opposing productions of the two proteins.

Activation of Eosinophils Is More Closely Linked with Interleukin-5 and Nitric Oxide Production than Tumor Necrosis Factor-α and Immunoglobulin E Levels

Patients were eligible if they had the following: (1) a documented clinical history of acute asthma, (2) positive results for sIgE test (aeroand/or food allergens according to the ImmunoCAP test), and (3) elevated serum total IgE (tIgE) levels to confirm allergy. The exclusion criteria were as follows: (1) medication history including corticosteroids or histamine antagonists, (2) concurrent systemic infection with fever (>37.5 ° C), or (3) chronic illness. The following parameters were measured: eosinophil parameters (ECP and ECP/Eo ratio), IgE-related parameters (sIgE score and tIgE level), cytokines (IL-5 and TNF-α), and inflammatory parameters [NOx metabolites and high-sensitivity C-reactive protein (hsCRP)]. The serum ECP concentration was measured using a solid-phase, 2-site chemiluminescent immunometric assay on an Immulite 2000 analyzer (Siemens Healthcare Diagnostics, Tarrytown, N.Y., USA). The cutoff limit for a positive serum ECP result was set at >19 μg/l on the basis of the manufacturer’s instruction. sIgE was measured for 2 major inhalant allergens (Dermatophagoides pteronyssinus and D. farinae) and 2 food allergens (egg white and cow’s milk) using a fluoroenzyme immunoassay system (ImmunoCAP 100, Phadia AB, Uppsala, Sweden). sIgE concentrations >0.35 kU/l Eosinophils contain several proinflammatory mediators and are implicated in the initiation of diverse inflammatory responses [1] . Eosinophil cationic protein (ECP) is produced in eosinophilic progenitors, stored in cytoplasmic granules of mature eosinophils, and released from activated eosinophils [2, 3] . The ratio of serum ECP concentrations to peripheral blood eosinophil counts (ECP/Eo ratio) reflects the number of ECP molecules per circulating eosinophil, which is regarded as a potential indicator of eosinophil activation [4, 5] . Few studies have closely examined the relationships between the activation of eosinophils and the production of nitric oxide (NOx) and cytokines. The present study investigated the associations between eosinophil activation and the levels of interleukin-5 (IL-5), tumor necrosis factor-α (TNF-α), NOx metabolites, and allergen-specific immunoglobulin E (sIgE) scores. A total of 112 patients with acute bronchial asthma (57 males and 55 females) aged 3.2–9.5 years (median 6) were investigated. Ageand gender-matched healthy subjects (n = 25) were evaluated as a control group, who had no evidence of allergic diseases and atopic sensitization based on skin-prick test results. Received: January 9, 2013 Accepted after revision: February 17, 2013 Published online: July 11, 2013

Analyzing Serum Eosinophil Cationic Protein in the Clinical Assessment of Chronic Rhinosinusitis

Eosinophil cationic protein (ECP) is a major granule-derived protein with cytotoxic activity found in eosinophils and has been known as a useful marker of allergic inflammation. In this study, we assessed the clinical significance of ECP in chronic inflammation of the nasal mucosa by evaluating the relationship between eosinophil activity and serum ECP concentration in a cohort of subjects with or without chronic rhinosinusitis (CRS) and allergic rhinitis (AR). We retrospectively reviewed the medical records of 492 subjects and analyzed eosinophil percentage in nasal smears, serum eosinophil counts, serum ECP concentrations, symptom scores, CT scores, the size of nasal polyp, and recurrence of CRS at follow-up. Elevated serum ECP concentration was closely related with higher eosinophil expression in all subject's nasal smears and sera. CRS subjects without AR had a higher percentage of immune cells that were eosinophils compared with control subjects and it was similar to subjects' with AR only or with both CRS and AR. CRS subjects without AR also had significantly higher serum ECP concentrations and eosinophil counts compared with control subjects. Additionally, serum ECP concentration was significantly correlated with CT scores, symptom scores, polyp size, and recurrence rate in CRS subjects without AR. Serum ECP concentration can be used as a marker of local and systemic eosinophil expression. We conjecture that although serum ECP elevation may not be specific in AR, analysis of serum ECP concentration could be particularly useful in estimating the progression and prognosis of CRS.

Tolerance to a new free amino acid-based formula in children with IgE or non-IgE-mediated cow’s milk allergy: a randomized controlled clinical trial

BackgroundAmino acid-based formulas (Aaf) are increasingly used in children with cow’s milk allergy (CMA). To be labeled hypoallergenic these formulas must demonstrate in clinical studies that they don’t provoke reactions in 90% of subjects with confirmed CMA with 95% confidence when given in prospective randomized, double-blind, placebo-controlled challenge (DBPCFC) trials. The majority of available safety data on Aaf derived from patients with IgE-mediated CMA. Considering substantial differences in the immunologic mechanism and clinical presentation of non-IgE-mediated CMA it’s important to investigate the hypoallergenicity of these formulas also in these patients. We prospectively assessed the tolerance to a new commercially available Aaf in children affected by IgE- or non-IgE-mediated CMA.MethodsConsecutive patients affected by IgE- or non-IgE-mediated CMA, aged ≤ 4 years, were enrolled. DBPCFC was carried out with increasing doses of the new Aaf (Sineall, Humana, Milan, Italy), using validated Aaf as placebo. Faecal concentrations of calprotectin (FC) and eosinophilic cationic protein (ECP) were monitored.ResultsSixty patients (44 male, 73.3%, median age 37, 95%CI 34.5–39.6 months, IgE-mediated CMA 29, 48.3%) were enrolled. At the diagnosis clinical symptoms were gastrointestinal (46.6%), cutaneous (36.6%), respiratory (23.3%), and systemic (10.0%). After DBPCFC with the new Aaf, no patient presented early or delayed clinical reactions. Faecal concentration of calprotectin and of ECP remained stable after the exposure to the new Aaf.ConclusionsThe new Aaf is well tolerated in children with IgE- or non-IgE-mediated CMA, and it could be used as a safe dietotherapy regimen for children with this condition.Trial registrationThe trial was registered in the ClinicalTrials.gov Protocol Registration System (ID number: NCT01622426).

Serum Polyfluoroalkyl Concentrations, Asthma Outcomes, and Immunological Markers in a Case–Control Study of Taiwanese Children

Background: Perfluorinated compounds (PFCs) are ubiquitous pollutants. Experimental data suggest that they may be associated with adverse health outcomes, including asthma. However, there is little supporting epidemiological evidence.Methods: A total of 231 asthmatic children and 225 nonasthmatic controls, all from northern Taiwan, were recruited in the Genetic and Biomarkers study for Childhood Asthma. Structure questionnaires were administered by face-to-face interview. Serum concentrations of 11 PFCs and levels of immunological markers were also measured. Associations of PFC quartiles with concentrations of immunological markers and asthma outcomes were estimated using multivariable regression models.Results: Nine PFCs were detectable in most children (≥ 84.4%), of which perfluorooctane sulfonate (PFOS) was the most abundant (median serum concentrations of 33.9 ng/mL in asthmatics and 28.9 ng/mL in controls). Adjusted odds ratios for asthma among those with the highest versus lowest quartile of PFC exposure ranged from 1.81 (95% CI: 1.02, 3.23) for the perfluorododecanoic acid (PFDoA) to 4.05 (95% CI: 2.21, 7.42) for perfluorooctanic acid (PFOA). PFOS, PFOA, and subsets of the other PFCs were positively associated with serum IgE concentrations, absolute eosinophil counts (AEC), eosinophilic cationic protein (ECP) concentrations, and asthma severity scores among asthmatics.Conclusions: This study suggests an association between PFC exposure and juvenile asthma. Because of widespread exposure to these chemicals, these findings may be of potential public health concern.

Efficacy of theophylline plus salmeterol/fluticasone propionate combination therapy in patients with asthma

To compare the efficacy of theophylline plus salmeterol/fluticasone propionate combination product (SFC) with SFC plus placebo in asthmatic patients. In this randomized, stratified, parallel-group study, 325 patients were randomized to receive either 200mg theophylline plus 50/250μg SFC or placebo tablet plus 50/250μg SFC twice daily for 24 weeks. Outcome variables included the level of asthma control (assessed by the Asthma Control Test) and the number of patients experiencing ≥1 exacerbations during the 24-week treatment period. Testing of lung function as well as measurement of the levels of inflammatory markers in induced sputum was performed. There were significantly fewer patients with ≥1 asthma exacerbation in the theophylline plus SFC group (29.6%) when compared with the SFC plus placebo group (46.9%) (p=0.004). Theophylline plus SFC improved the FEF(25-75%) value, which indicates enhanced small airway function, to a greater extent than SFC plus placebo (66.9±18.8% and 57.4±17.6%, respectively; p<0.001). A significant decrease in eosinophil count and concentration of eosinophil cationic protein in induced sputum was also seen in the theophylline plus SFC group when compared with the SFC plus placebo group (4.1±2.2% and 6.3±2.7%, 63.6±39.5μg/L and 89.4±45.6μg/L, respectively; all p<0.01). The combination of theophylline plus SFC may provide greater protection against asthma exacerbations, and its administration is accompanied by significant improvements in small airway function and airway inflammation.