Eosinophil Cationic Protein Concentrations Research Articles

Fungal allergy as a risk factor for developing rosacea

The pathogenesis of rosacea is not fully understood. In this disease, the release of inflammatory molecules and overgrowth of skin-colonizing microorganisms, including fungal microbiota, are noted. The study of the frequency of occurrence and the nature of the spectrum of sensitization to fungal allergens in patients with rosacea is of particular interest and accounts for relevance of the study. Purpose of the study: to analyze the frequency of occurrence and the nature of the spectrum of sensitization to fungal allergens in patients with rosacea.
 Materials and methods. The study included patients with rosacea aged 18 to 60 years (group 1, n = 31). All patients underwent a specific allergic examination: collected allergic history, skin prick testing using the following allergens: nutritional yeast, Candida albicans, Alternaria alternate, Aspergillus fumigatus, Cladosporium herbarum, Penicillium notatum (Allergopharma, Germany). Concentration of total immunoglobulin E (IgE) and eosinophilic cationic protein in blood serum was measured in indirect immunofluorescence analysis with Thermo Scientific Multiskan FC semi-automatic analyzer. Statistica 8.0 software package was used for statistical analysis.
 Results. Analysis of the features of the spectrum of sensitization to fungal allergens in the group of patients with rosacea showed that the most significant allergens are Cladosporium herbarum (74.2% of patients) and Candida albicans (58.1%). Sensitization to nutritional yeast and Penicillium notatum was detected in almost half of the patients. The concentration of total immunoglobulin E in blood serum in the group of patients with rosacea was 79.4 IU/ml [23.4; 105.1], in the control group 45.1 IU/ml [23.4; 144.0]. The concentration of eosinophilic cationic protein in blood serum in the group of patients with rosacea was significantly higher compared to the control group: 50.3 ng/ml [43.0; 62.0] and 7.9 ng/ml [4.6; 27.1], respectively. After skin prick testing with fungal allergens, patients with rosacea were prescribed an individual elimination diet with the exclusion of causally significant allergens for a period of 1 month. In 61.2% (n = 19) of cases in patients with rosacea, a positive elimination effect was noted: a significant decrease in the severity of erythema, papules and pustules in the face.
 Conclusion. Thus, the study revealed a high incidence of fungal allergy and established the features of the spectrum of sensitization to fungal allergens in patients with rosacea. The most significant fungal allergens in rosacea are fungi of the genus Cladosporium herbarum, Candida albicans, Penicillium notatum as well as nutritional yeast.

Cationic proteins from eosinophils bind bone morphogenetic protein receptors promoting vascular calcification and atherogenesis.

Blood eosinophil count and eosinophil cationic protein (ECP) concentration are risk factors of cardiovascular diseases. This study tested whether and how eosinophils and ECP contribute to vascular calcification and atherogenesis. Immunostaining revealed eosinophil accumulation in human and mouse atherosclerotic lesions. Eosinophil deficiency in ΔdblGATA mice slowed atherogenesis with increased lesion smooth muscle cell (SMC) content and reduced calcification. This protection in ΔdblGATA mice was muted when mice received donor eosinophils from wild-type (WT), Il4-/-, and Il13-/- mice or mouse eosinophil-associated-ribonuclease-1 (mEar1), a murine homologue of ECP. Eosinophils or mEar1 but not interleukin (IL) 4 or IL13 increased the calcification of SMC from WT mice but not those from Runt-related transcription factor-2 (Runx2) knockout mice. Immunoblot analyses showed that eosinophils and mEar1 activated Smad-1/5/8 but did not affect Smad-2/3 activation or expression of bone morphogenetic protein receptors (BMPR-1A/1B/2) or transforming growth factor (TGF)-β receptors (TGFBR1/2) in SMC from WT and Runx2 knockout mice. Immunoprecipitation showed that mEar1 formed immune complexes with BMPR-1A/1B but not TGFBR1/2. Immunofluorescence double-staining, ligand binding, and Scatchard plot analysis demonstrated that mEar1 bound to BMPR-1A and BMPR-1B with similar affinity. Likewise, human ECP and eosinophil-derived neurotoxin (EDN) also bound to BMPR-1A/1B on human vascular SMC and promoted SMC osteogenic differentiation. In a cohort of 5864 men from the Danish Cardiovascular Screening trial and its subpopulation of 394 participants, blood eosinophil counts and ECP levels correlated with the calcification scores of different arterial segments from coronary arteries to iliac arteries. Eosinophils release cationic proteins that can promote SMC calcification and atherogenesis using the BMPR-1A/1B-Smad-1/5/8-Runx2 signalling pathway.

Predictive Value of Eosinophil Cationic Protein in Nasal Secretions in Eosinophilic Chronic Rhinosinusitis.

To investigate the value of secretions Eosinophilic cationic protein (ECP) detection in the diagnosis of endotypes of Chronic rhinosinusitis (CRS) and its correlation with clinical symptoms, so as to provide guidance for the clinical application of EOS and ECP detection in secretions. Patients' nasal secretions and polyps (or middle turbinate for control) were collected and their EOS% and ECP levels were measured. Correlation analysis was performed for EOS% and ECP levels in secretions and tissues, respectively. The correlation between secretions EOS% and ECP and clinical symptom scores (symptomatic visual analog scale (VAS) scores, Lanza-kennedy scores from nasal endoscopy and Lund-Mackay scores from sinus CT) was further analyzed. Receiver operating characteristic curves were used to assess the predictive potential of EOS% and ECP in nasal secretions. Eosinophilic chronic rhinosinusitis (ECRS) patients had higher concentrations of ECP in nasal secretions than healthy subjects and NECRS (non-eosinophilic CRS) (p < 0.0001;0.0001); EOS% in nasal secretions was higher in ECRS than healthy subjects (p = 0.0055), but the differences between ECRS and NECRS were not statistically significant (p = 0.0999). Correlation analysis showed that tissue EOS% was correlated with ECP concentration and EOS% in nasal secretions (R = 0.5943;0.2815). There was a correlation between EOS% in secretions with a total LM score (R = 0.3131); ECP concentration in secretions with a total LK score (R = 0.3792). To diagnose ECRS, the highest area under the curve (0.8230) was determined for ECP in secretions; the highest area under the curve (0.6635) was determined for EOS% in secretions. Measurement of ECP in nasal secretions is useful for non-invasive diagnosis of ECRS. 3 Laryngoscope, 133:3304-3312, 2023.

Eosinophil Granule Proteins Involvement in Acute Appendicitis-An Allergic Disease?

Several pieces of evidence point to an allergic component as a trigger of acute appendicitis. As the Th2 immune response is characterized by eosinophil mobilization to the target organ and release of their cationic granule proteins, it is reasonable to investigate if the degranulation of eosinophils could be associated with the local injury. The primary aim of this study is to evaluate the participation of eosinophils granules proteins in acute appendicitis, both at local and systemic levels and the secondary aim is to evaluate the diagnostic accuracy of eosinophils granules proteins for the detection of acute appendicitis, as well as for distinguishing between complicated and uncomplicated acute appendicitis. Eosinophil-derived neurotoxin (EDN), eosinophil cationic protein (ECP) and eosinophil peroxidase (EP) are the most well-known eosinophil granule proteins. From August 2021 to April 2022, we present a prospective single-center study to evaluate the EDN, ECP, and EP concentrations simultaneously in appendicular lavage fluid (ALF) and the serum of 22 patients with acute phlegmonous appendicitis (APA), 24 with acute gangrenous appendicitis (AGA), and 14 normal controls. Concerning EDN, no differences were found between groups. ECP concentrations in ALF and serum were significantly higher in the histologically confirmed acute appendicitis compared to the control groups (p < 0.0001 and p < 0.0001, respectively). In ALF, no differences were found between ECP levels in APA: 38.85 ng/mL (IQR 26.50-51.77) and AGA 51.55 ng/mL (IQR 39.55-70.09) groups (p = 0.176). In the serum, no difference was found between ECP levels at APA: 39 ng/mL (IQR 21.30-56.90) and AGA: 51.30 ng/mL (IQR 20.25-62.59) (p = 0.100). For EP, the concentrations in ALF (p < 0.001) and serum (p < 0.001) were both higher in acute appendicitis compared to the control. In ALF, no difference was found between APA: 240.28 ng/mL (IQR 191.2-341.3) and AGA: 302.5 (IQR 227.7-535.85) (p = 0.236). In the serum, no differences were found between APA: 158.4 ng/mL (IQR 111.09-222.1) and AGA: 235.27 (IQR 192.33-262.51) (p = 0.179). Globally, the ALF concentrations were higher than serum concentrations, reflecting an intense inflammatory local reaction in AA. The optimal ECP cut-off for discriminating between acute appendicitis and the controls was >11.41 ng/mL, with a sensitivity of 93.5%, but with a specificity for identifying appendicitis of 21.4%, good discriminative power (AUC = 0.880). For EP, the optimal cut-off was >93.20 ng/mL, with a sensitivity of 87%, but with a specificity of 14.3% (AUC = 0.901), excellent discriminative power. For the diagnosis of perforated AA, the discriminative power of ECP and EP serum concentrations are weak (AUC = 0.562 and AUC = 0.664, respectively). Concerning the presence of peritonitis, the discriminative power of ECP and EP serum concentrations is acceptable, respectively: AUC = 0.724 and AUC = 0.735. Serum levels of EDN (p = 0.119), ECP (p = 0.586) and EP (p = 0.08) in complicated appendicitis were similar to uncomplicated appendicitis. Serum concentrations of ECP and EP can be added to decision-making AA diagnosis. A Th2-type immune response is present in AA. These data bring forward the role of an allergic reaction in the pathogenesis of acute appendicitis.

Eosinophilic myocarditis: comprehensive update on pathophysiology, diagnosis, prognosis and management.

Hypereosinophilic syndromes are a group of disorders secondary to the accumulation of eosinophils leading to the injury of one or more organs. Among them, eosinophilic myocarditis (EM) is a rare form of inflammatory cardiomyopathy characterized by eosinophilic infiltration into myocardial tissue and subsequent release of substances with cell membrane damage and cell destruction. The degree of infiltration is thought to depend on the underlying condition, as well as the degree and duration of eosinophil exposure and ranges from mild localized disease to diffuse multifocal infiltrates associated with myocardial necrosis, thrombotic complications and endomyocardial fibrosis. The main causes of EM are hypersensitivity reactions, eosinophilic granulomatosis with polyangiitis, hypereosinophilic syndrome variants, infections and cancer. Clinical presentation can be variable, ranging from asymptomatic forms to life-threatening conditions, to chronic heart failure due to progression to chronic restrictive cardiomyopathy. Marked eosinophilia in peripheral blood, elevated serum eosinophilic cationic protein concentration and multimodality imaging may suggest the etiology of EM, but in most cases an endomyocardial biopsy must be performed to establish a definitive diagnosis. Systemic treatment varies greatly depending on the underlying cause, however the evidence of an eosinophilic infiltrate allows initiation of immunosuppressive therapy, which is the mainstay of treatment in idiopathic and in most forms of EM. Patients with helminthic infection benefit from anti-parasitic therapy, those with myeloid clone often need a tyrosine kinase inhibitor, while anticoagulant therapy should be undertaken in case of possible thrombotic complications.

Inflammatory mediators in nasal secretions of patients with nasal polyposis with and without aspirin sensitivity

The aim of this cross-sectional study was to compare the levels of inflammatory mediators in nasal secretions in patients with aspirin-exacerbated respiratory disease (AERD) and in those with nasal polyposis (NP) without aspirin-sensitivity and to correlate nasal fluid mediator concentrations with clinical parameters of the disease. A total of 30 patients with AERD, 30 chronic rhinosinusitis (CRS) with NP patients without aspirin sensitivity (CRSwNP), and 30 control subjects without inflammation of the nasal mucosa (C), selected for surgical treatment entered the study. The total nasal symptom score (TNSS), endoscopic score (ES), and Lund-Mackay score (LMS), were evaluated. The concentrations of eosinophil cationic protein (ECP), tryptase, heat shock protein 70 (HSP70), substance P and Clara cell protein 16 (CC16) were determined in nasal secretions. Higher concentrations of ECP, tryptase, and HSP70 were measured in the AERD patients than in the CRSwNP patients and the C group (p < .001; p < .001, respectively for all mediators). However, levels of CC16 were higher in the C group than in the AERD and CRSwNP groups (p < .001; p < .001, respectively). A positive correlation between the TNSS and CC16 and a negative one between CC16 and tryptase levels were found in the C group. The CRSwNP group showed positive correlations between ECP, HSP70, and tryptase and negative correlations between substance P, ES, and LMS, as well as between CC16 and tryptase levels. In the AERD group, we found a positive correlation between HSP70 and ECP levels and a negative correlation between the TNSS and CC16 concentration. The obtained results indicate the increased production of mediators of eosinophil and mast cell function, and the decreased production of biomarker of respiratory epithelial function in AERD patients. Clinical and biochemical parameters correlate in different ways in the AERD and CRSwNP patients.

INCREASED EOSINOPHIL GRANULE PROTEIN PRODUCTION IN CHRONIC SPONTANEOUS URTICARIA

<h3>Introduction</h3> Chronic spontaneous urticaria (CSU) lesional and non-lesional skin reveals increased numbers of eosinophils, but the exact role of eosinophils in CSU pathogenesis remains unknown. A pathogenic role for eosinophils in CSU was supported by the results of a recent clinical trial demonstrating the efficacy of benralizumab in CSU. Previous studies have identified increased levels of major basic protein (MBP) and eosinophilic cationic protein (ECP) in the sera and skin of CSU subjects; however, eosinophil peroxidase (EPO), the granule protein uniquely expressed by eosinophils, and eosinophil-derived neurotoxin (EDN) were not examined. Therefore, we sought to determine whether eosinophil granule proteins (EGPs) are increased in CSU. <h3>Methods</h3> Sera from CSU subjects (n=11) and healthy controls (n=1) were collected and sent to the NIH Human Eosinophil Section for analysis with their novel multiplex immunoassay which simultaneously measures concentrations of MBP, ECP, EDN, and EPO using Luminex xMAP technology. CSU subjects completed disease activity surveys and complete blood count differentials were analyzed. <h3>Results</h3> Mean concentrations of MBP (1329.23 vs 399.55ng/ml), ECP (285.87 vs 45.31ng/ml), EDN (349.43 vs 38.53ng/ml), and EPO (54.96ng/ml vs undetectable) were markedly increased in CSU subjects compared to the healthy control. We found no correlation between eosinophil counts and granule protein levels, or with Urticaria Activity Scores. <h3>Conclusions</h3> Our results suggest that unique pathways may be activated in a subset of patients with CSU which led to the production of large quantities of EGPs. Additional research is needed to understand the pathways responsible for producing these granule proteins in CSU and their clinical impact.

Clinical and molecular features of virus-induced acute exacerbations of chronic obstructive pulmonary diseas

Introduction. Inflammation in viral-induced acute exacerbations of chronic obstructive pulmonary disease (COPD) is not studied enough.The aim was to establish molecular pattern of inflammation in viral-induced acute exacerbations of chronic obstructive pulmonary disease (AECOPD) in comparison with bacterial AECOPD and to reveal associations with AECOPD phenotype and subsequent COPD progression.Materials and methods. Subjects hospitalized with acute exacerbations of COPD (AECOPD) of which 60 were viral, 60 were bacterial and 60 were viral-bacterial were recruited to single center prospective (52 weeks) cohort study. Control group – 30 healthy people. COPD were diagnosed previously during stable phase of the disease according to spirographic criteria. Viral AECOPD were confirmed by detection of RNA of influenza A and B, respiratory syncytial virus, rhinovirus or severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in sputum or bronchoalveolar lavage fluid (BALF) using reverse transcription-polymerase chain reaction (RT-PCR). Bacterial AECOPD were confirmed by sputum/BALF neutrophilia or elevated blood procalcitonin levels or by detecting bacteria by standard culture method. Plasma concentrations of cytokines, fibrotic markers, enzymes were measured by enzyme-linked immunosorbent assay, plasma fibrinogen – by Clauss method. Complex lung function investigation, Dopplerechocardiography, subsequent AECOPD assessment were done. Kruskal-Wallis and chi-square test were used to compare groups, Cox regression and linear regression – to explore relationships.Results. Viral AECOPD were characterized by highest plasma concentrations of Eosinophilic cationic protein (62,3 (52,4; 71,0) ng/ml)), interleukin-5 (IL-5) (11,3 (8,4; 15,9) pg/ml), fibroblast growth factor-2 (FGF-2) (10,4 (6,2; 14,9) pg/ml), transforming growth factor-β1 (TGF-β1) (922,4 (875,7; 953,8) pg/ml), hyaluronic acid (185,4 (172,8; 196,3) ng/ml), amino-terminal propeptide of type III procollagen (PIIINP) (249,2 (225,1; 263,7) ng/ml), matrix metalloproteinase-1 (MMP-1) (235,2 (208,6; 254,9) pg/ml). Levels of IL-5 during AE COPD was the predictor of FEV1, bronchodilation coefficient, subsequent exacerbations at remote period, fibrinogen was associated with FEV1, PIIINP and FGF-2 with DLco, PaO2, mean pulmonary artery pressure (mPAP), exacerbations, MMP-1 – with mPAP.Conclusions. In virus-induced AECOPD inflammation pattern differed from those in bacterial one and associated with AECOPD phenotype and COPD phenotype at the stable phase.

Study the Role of Eosinophilic Cationic Protein, Copper and Iron in Women Infected with Trichomonas vaginalis Parasite

Since Trichomonas vaginalis (T. vaginalis) only manifests as a trophozoite stage during sex, it is typically transmitted during sex as a trophozoite. The purpose of this study was to determine the concentrations of copper, iron, and Eosinophil cationic protein (ECP) in vaginitis patients. Between July and November 2021, 355 suspected patients who visited the AL-Zahra maternity and pediatrics laboratory in the AL-Najaf region (Iraq) and 30 healthy respondents of the same age participated in the study. According to the manufacturer's instructions, the ELISA technique was used to measure the levels of three biomarkers in serum (copper, iron, and ECP). On the other hand, the colorimetric approach was used to measure the copper concentration. The investigation showed that in the blood sample of T. vaginalis-infected patients, the amounts of copper and iron considerably decreased (P &gt; 0.05). In contrast, the serum levels of ECP considerably increased (P &gt; 0.05) when compared to the control group. As a result of the T. vaginalis infection, copper levels in the blood are dramatically impacted, according to the current study's findings. The current study results suggested that the T. vaginalis infection alters serum iron and ECP concentrations, significantly affecting copper levels in the blood.

Clinical significance of sensitization to house dust mite allergens using flow cytometry-based basophil activation test

Respiratory allergy takes a significant place among allergic diseases. Similar clinical manifestations occur in sensitized patients in response to various airborne allergens. Accordingly, the allergen testing by means of in vivo and in vitro techniques is necessary to identify a causally significant allergen. The main laboratory assay to estimate sensitization to the airborne allergens is based on detection of specific immunoglobulin E (sIgE). However, clinical manifestations cannot be always confirmed by this test. The aim of this work was to evaluate the potential of basophil activation test (BAT) using flow cytometry, in order to confirm sensitization to house dust mite allergens. The BAT was carried out with allergens from Dermatofagoides in 34 volunteers, 130 patients were examined at the Russian Centre of Emergency and Radiation Medicine (33 people with household and 97 people with polysensitization), and 10 patients with allergy to house dust mites confirmed by skin testing. The sensitization was assessed using two laboratory kits (Allergenicity and BD FastImmune). The total IgE and sIgE to house dust mites, as well as concentrations of eosinophilic cationic protein, were determined. Specificity of the BAT with Allergenicity kit for the house dust mites was 94%. The sensitivity was 88% in the patients with household sensitization, thus being comparable with skin testing results available from the literature. An absolute correlation was found between positive results of skin testing and the BAT. In patients with household sensitization, sIgE levels &gt; 0.35 UA/mL were detected in 39% of cases. A positive correlation was shown between the indices of basophil activation in response to testing allergens, and total immunoglobulin E and sIgE concentrations. High degree of correlation was found between the results of Dermatofagoides sensitization testing with both BAT kits. It was shown that the use of H1 histamine receptor blockers had no effect on BAT results. A decrease in sensitivity assay for airborne allergens was revealed upon therapy with glucocorticosteroids. Due to the proven clinical significance of the basophil activation test for diagnosing sensitization to inhaled allergens, one may require to supplement the range of approved assays in Russian Clinical Guidelines with BAT testing to prove sensitization in patients with allergic rhinitis and bronchial asthma, thus contributing to better diagnosis of causally significant allergens and, accordingly, administration of proper therapy.

INFLAMATOMETRIC FEATURES OF BRONCHIAL ASTHMA WITH DIFFERENT ACTIVITY DEGREE OF EOSINOPHILIC AIRWAY INFLAMMATION

The aim of the study – to investigate the clinical signifi cance of eosinophilic cationic proteinconcentration in sputum and its diagnostic role in the disease management for optimizationof the personalized anti-infl ammatory therapy of patients with bronchial asthma.&#x0D; Materials and methods. There have been examined 66 school-aged children with persistentbronchial asthma who had been under anti-infl ammatory control therapy for at least thelast three months, but still needed treatment correction. Sputum has been collected in allpatients during post-attack period of disease by the method of sputum induction usingan inhalation of hypertonic solutions (3%,5%,7%) of sodium chloride. Quantitative andqualitative cytological composition of the sputum precipitate has been determined, inthe supernatant liquid fraction, obtained after sputum centrifugation, the concentrationsof eosinophilic cationic protein (ECP) and endothelial vascular growth factor (VEGF)have been studied by ELISA method. In the general cohort of examined patients, theaverage ECP concentration was 2.28±2.2 ng/ml (minimum value 0, maximum – 9.2 ng/ml). Depending on the ECP content in the sputum, children have been divided into 2clinical groups: The fi rst (I) clinical group was formed by 29 children with a content ofECP in the sputum greater than the average cohort value, and the second (II) clinicalgroup included remaining 47 patients with the level of ECP&lt;2 ng/ml). According to themain clinical characteristics (age, gender, place of residence, duration of the disease)the clinical groups have been comparable. All patients underwent a biochemical studyof exhaled breath condensate (EBC), which involved the content determination of totalprotein, catalase activity; concentrations of nitrogen monoxide metabolites and markers&#x0D; of proteolytic activity. The results of the study were analyzed by parametric and non-parametric calculation methods, and estimation of the tests’ diagnostic value were carried&#x0D; out from the position of clinical epidemiology taking into account their sensitivity (Se)and specifi city (Sp), as well as attributive (AR) and relative (RR) risks, and the odd ratio(OR) of the event, taking into consideration their 95% confi dence intervals (95% CI).Results. The analysis of the obtained data has showed that in the comparison groups theclinical level of asthma control (18.3 ± 1.5 vs. 18.6 ± 1.3 points, P&gt; 0.05), and sputumcell composition (in particular by the number of eosinophilic granulocytes: 9,24±2.3vs. 9,28±2.2%, P&gt; 0.05) did not diff er much. In addition to that, VEGF concentrationin sputum, which refl ects the bronchial remodeling processes, was 1.25 times higherin representatives of the I group (145.43 ± 19.71 pg/ml) compared with patients of theII group (115.93 ± 15, 7 pg/ml; P&gt; 0.05). In EBC, the content of nitrogen monoxidemetabolites was higher in patients of the II group: 44.4 ± 5.74 μmol/l in comparison withrepresenatives of I group (55.8 ± 9.8 μmol/l; P&gt; 0.05). At the same time, in patients ofclinical I group in EBC higher proteolytic activity of azocol, as well as a higher catalaseactivity was noted (68.4 ± 22.6 μmol/min x mg of protein compared to 49.6 ± 10.3 μmol/min x mg of protein in the II group of comparison; P&gt; 0, 05). In contrast, in the II group,the infl ammatory process has been associated with the lysis of low molecular weightproteins.Conclusion. In children with bronchial asthma, increased degranulation of sputumeosinophils (according to the ECP concentration) is associated with a more pronouncedairway infl ammation, as evidenced by the increased intensity of oxidative modifi cation ofproteins (OR=3,6) and proteolytic activity of high molecular weight proteins; togetherwith high catalase activity and elevated concentration of such marker of bronchialremodeling as VEGF, along with decreased content of nitrogen monoxide metabolites inthe exhaled breath condensate.

Diagnostic value of a novel test paper detection of ECP in nasal secretion for allergic rhinitis

Objective:To explore the diagnostic value of a novel test paper, which detect eosinophil cationic protein（ECP） of nasal secretion in allergic rhinitis（AR）. Methods:Nasal secretion and serum samples from 107 patients with allergic rhinitis（AR group） and 40 healthy volunteers（control group） were selected. The nasal symptoms were also evaluated in AR group. The degree of ECP coloration was evaluated by nasal secretion eosinophil cationic protein-myeloperoxid（ECP-MPO） test paper, and the concentration of ECP in nasal secretion and the concentration of cytokines in serum were detected at the same time. The difference and correlation among these indexes were analyzed. The best cutoff value and test efficiency of ECP chromogenic grade and concentration of nasal secretion were calculated by receiver operating characteristic curve（ROC）. Results:The concentration of ECP in nasal secretion of AR patients was significantly higher than that of healthy controls（P<0.05）. The color grade of nasal secretion detected by the test paper was positively correlated with the concentration of ECP in nasal secretion（P<0.05）, and there was significant difference among different grades（P<0.05）. There was a satisfying symmetry between the ECP color grade of nasal secretion and the serum specific IgE（sIgE） level as well as a high diagnostic consistency between them（P<0.05）. The area under the curve（AUC） of ECP concentration ROC in nasal secretion was 0.807 2, corresponding to 64% sensitivity and 85% specificity when the cutoff value was set at 0.980 5; when the cutoff value was set at 1, the AUC of nasal secretion ECP color grading was 0.941 9, corresponding to 92% sensitivity and 94% specificity. No clear correlation between the concentration of ECP in nasal secretion and serum cytokines was found（P>0.05）. Conclusion:The results of this novel test paper is in good agreement with those of serological allergens. It could serve as a preliminary test to evaluate the severity of allergy with satisfactory sensitivity and specificity, and is especially suitable in clinical practice for primary hospital.

Increased fecal human beta-defensin-2 expression in preterm infants is associated with allergic disease development in early childhood

BackgroundThis study aimed to investigate whether fecal human beta-defensins (HBD)-2 and eosinophil cationic protein (ECP) expression in preterm infants are associated with allergic disease development by age 2 years. MethodsPreterm infants' stool samples were collected at the age of 6 and 12 months postnatally. Information regarding medication exposure histories (antibiotics, antipyretics, probiotics) and physician-diagnosed allergic diseases was obtained using age-specific questionnaires and medical records. We compared the 6-month and 12-month fecal HBD-2 and ECP concentrations between the medication exposure and non-exposure group, respectively, and between children who developed allergic diseases and those who did not by 2 years of age. Univariate and multivariable logistic regression analyses were performed to investigate independent variables related to physician-diagnosed allergic diseases by 2 years of age. ResultsSeventy-four preterm infants (gestational age, 31–36 weeks) were included. Fecal HBD-2 levels were significantly increased at 12 months of age among children who developed allergic diseases compared to those who did not (37.18 ± 11.80 ng/g vs. 8.56 ± 4.33 ng/g, P = 0.011). This association was more apparent among allergic children given antibiotics (50.23 ± 16.15 ng/g vs. 9.75 ± 7.16 ng/g, P = 0.008) or antipyretics (46.12 ± 14.22 ng/g vs. 10.82 ± 6.81 ng/g, P = 0.018) during the first year, whereas among allergic children who were previously not exposed to antibiotics or antipyretics, the differences were not significant. Results of the multivariable logistic regression analysis indicated that HBD-2 concentration in 12-month stools was an independent indicator associated with physician-diagnosed allergic diseases by 2 years of age (adjusted odds ratio: 1.03 [95% confidence interval: 1.00–1.05], P = 0.036). Our data revealed a lack of association between fecal ECP and allergic diseases. ConclusionsWe found that preterm infants who expressed high fecal HBD-2 at 12 months of age were associated with physician-diagnosed allergic diseases by the age of 2 years. Further studies are needed to determine the role of fecal HBD-2 in the development of allergic diseases.

Different levels of mucus inflammatory mediators in nasal polyposis with and without aeroallergen sensitivity.

ObjectivesBiomarker levels in nasal secretions can reflect the inflammatory status of nasal mucosa and evolution of sinus disease. The aim of this study was to evaluate the relationship between local inflammatory mediator production and clinical characteristics of patients with nasal polyposis (NP).MethodsThirty‐one nonaeroallergen sensitized patients with NP (NANP), 29 aeroallergen sensitized patients with NP (ANP), and 30 subjects without inflammation of nasal mucosa as controls (C) entered this prospective, cross‐sectional study. Clinical parameters (symptoms, endoscopic, and radiological findings) were assessed. The concentrations of heat shock protein 70 (HSP70), eosinophil cationic protein (ECP), tryptase, substance P and Clara cell protein 16 (CC16) were measured in the nasal secretion samples of all participants by ELISA method.ResultsOur results showed higher concentrations of HSP70, ECP, and tryptase in ANP than in NANP and C (p < .001 for all markers). On the other hand, levels of CC16 were significantly higher in C than in NANP and ANP groups (p < .001; p < .001, respectively). We found positive correlations between HSP70, ECP, tryptase, and substance P levels and nasal symptom score in patients with NP. Also, HSP70, ECP, tryptase, and substance P showed different levels of positive correlation among themselves, with HSP70 showing highest positive correlation with ECP. Finally, relatively strong negative correlations were found between the levels of CC16 and nasal symptoms, as well as between the CC16 levels and levels of other four mediators in nasal fluid.ConclusionHSP70, ECP, tryptase, and substance P might play a role in the pathogenesis of NP. The results suggest that chronic inflammation in NP involves a self‐sustaining local release of HSP70, ECP, and tryptase, independent of aeroallergen stimulation of the mucosal layer, although the production of these mediators is higher in aeroallergen sensitized NP patients.

Microvessel quantification by fully convolutional neural networks associated with type 2 inflammation in chronic rhinosinusitis

BackgroundThe pathogenesis of chronic rhinosinusitis (CRS) is still unclear, and little is known about angiogenesis in this disease. We utilized a fully convolutional network (FCN), which has been extensively used in image processing to study angiogenesis in CRS. ObjectiveTo explore the tissue quantification of microvessels and their potential association with inflammation in CRS by using FCN to reflect the angiogenesis condition in CRS. MethodsFor endotyping of CRS, tissue homogenates of 79 patients with CRS who had undergone functional endoscopic sinus surgery and 17 control subjects were analyzed for interferon gamma, transforming growth factor beta, interleukin (IL)-1β, IL-5, IL-6, IL-8, IL-10, IL-17, tumor necrosis factor alpha, eosinophilic cationic protein, immunoglobulin E, and Staphylococcus aureus-immunoglobulin E(SE-IgE). A total of 552 hematoxylin and eosin-stained images of 27 CRS tissue samples were used to develop an FCN, going through training, validation, and evaluation processes. An optimized FCN was applied to quantify the microvessels of tissue samples of all subjects. Correlation analysis between microvessel quantification with phenotype, endotype, clinical characteristics, and cytokine expression of CRS was carried out. ResultsQuantification of microvessels in type 2 and non-type 2 CRS demonstrated considerable differences, with a higher expression in type 2 CRS. There was a strong negative correlation between the area ratio of microvessels with tissue tumor necrosis factor alpha and transforming growth factor beta levels and a mildly positive correlation with tissue IL-5 and eosinophilic cationic protein concentration. ConclusionFCN proved to facilitate the analysis of microvessels in airway tissue samples. This study elucidated the close association of angiogenesis with endotyping, suggesting that treatment aiming at antagonizing angiogenesis may assist to the therapy for the recrudescent and refractory CRS.

Effect of IL-35 on apoptosis, adhesion, migration, and activation of eosinophils in allergic rhinitis.

Eosinophils play critical roles in the development of allergic rhinitis (AR) by releasing toxic substance. Interleukin-35 (IL-35), a newly identified anti-inflammatory cytokine, had potent inhibitive role for eosinophil infiltration in allergic disease. However, the direct effect of IL-35 on eosinophil was not clear. Twenty AR children and sixteen controls were recruited. The correlation between IL-35 protein expression and blood eosinophil counts and activation was analyzed. The effect of IL-35 on eosinophil apoptosis and adhesion was analyzed by flow cytometry. Transwell system was used for the migration assay. The eosinophil cationicprotein (ECP) from supernatant of eosinophils after IL-35stimulation was detected by enzyme-linked immunosorbent assay kits. The IL-35 protein levels were negatively correlated with eosinophil counts (p<.01) and ECP concentration (p<.01) in AR children. IL-35 promotes apoptosis and inhibits adhesion, migration, and activation of eosinophils. Moreover, the mRNA expression of IL-12 receptor β2 and glycoprotein 130 were significantly enhanced by eosinophils after IL-35stimulation. The apoptosis induced by IL-35 was mediated by phosphoinositide 3-kinase (PI3K) pathway. IL-35 inhibits adhesion of eosinophils through extracellular regulated proteinkinases (ERK) and PI3K pathways. The eosinophil chemotaxis and activation affected by IL-35 were mediated by PI3K and p38mitogen-activated proteinkinase (MAPK) pathways. Our results confirmed that IL-35 played inhibitive roles in apoptosis, adhesion, migration, and activation of eosinophils in AR, implying that IL-35may be used as treatment target in future.

Personalized approach to diagnostics and therapy of patients with psoriasis

Establishing causal relationship between allergy and psoriasis and determining appropriate sensitization profile may guide clinicians in terms of specific allergological diagnostics and personalized therapy. Our purpose was to study the occurrence of food allergy and certain sensitization profile to food allergens in patients with psoriasis, in order to establish new approaches to specific diagnosis and treatment of the disorder.The study involved patients with psoriasis vulgaris (n = 51) aged 18 to 66 years. The control group included practically healthy people matched by sex and age (n = 19). All the patients underwent allergological examination: collection of allergic history, determination of total immunoglobulin E (IgE) and eosinophilic cationic protein concentration in blood serum by means of indirect immunofluorescence analysis using Thermo Scientific Multiskan FC semi-automatic analyzer. The sensitization profiles to food and pollen allergens were also studied. Calculation and analysis of these data was carried out using Statistica 8.0 software package.Concentration of total immunoglobulin E in blood serum in the group of patients with psoriasis was 57.9 IU/ml (31.6-135.1) compared to control group, with 45.1 IU/ml (23.4-144.0). The levels of serum eosinophilic cationic protein was 8.6 ng/ml (4.6-20.3) in the patients with psoriasis, being 7.9 ng/ml (4.6-27.1) in controls. When studying the spectrum of sensitization to food allergens in patients with psoriasis, some specific features were revealed. E.g., sensitization to cow’s milk was detected in 33.3% (n = 17); to beef, in 39.2% of cases (n = 20), to whole chicken egg – 29.4% (n = 15), to chicken, in 39.2% (n = 20); to wheat flour, 37.2% (n = 19); to rye flour, in 31.4% (n = 16); to rice and buckwheat, 21.6% (n = 11), to yeast, in 23.5% of the patients (n = 12). Upon administration of individual elimination diet for 1 month, we observed a decrease or complete regression of itching, decreased severity of infiltration, hyperemia and peeling in the foci of psoriatic skin lesions in 68% of the cases. Normal concentration of total immunoglobulin E and eosinophilic cationic protein in blood serum of patients with psoriasis combined with positive results of skin-prick tests and elimination effect presumes a participation of non-atopic immune pathologies when triggering allergy in psoriatic disease.

Eosinophilic cationic protein (ECP) in the clinical work-up of chronic cough.

Chronic cough is a common symptom, addressed in the clinical setting by empirical treatment together with some laboratory investigations. The aim of the present study was to investigate the value of testing eosinophilic cationic protein (ECP) serum levels combined with other diagnostic procedures and empirical treatment in the diagnostic workup of chronic cough. In this study, we evaluated 194 patients with chronic cough. No subject had received any anti-inflammatory treatment before clinical evaluation, and none was an active smoker. ECP was measured with a commercially available fluoroenzyme immunoassay and results were expressed as µg/L. The analysis of variance showed that mean ECP level differs among the various diagnosis categories (P<0.001). Mean ECP level was significantly higher in asthmatic patients, particularly in the active disease. Serum ECP concentration could represent a useful biomarker in the clinical work-up of chronic cough, managing to differentiate asthma from other chronic disorders.

Effects of Medication Use on Small Airway Function and Airway Inflammation in Patients with Clinically Controlled Asthma

To observe effects of medication use on small airway function, airway inflammation and acute exacerbations in patients with clinically controlled asthma. Forced expiratory flow over the middle half of the forced expiratory curve (FEF25%-75%), percentage of eosinophil, concentrations of eosinophil cationic protein (ECP) and interleukin (IL)-5 in induced sputum were assessed in patients with clinically controlled asthma who were given oral anti-inflammatory agents alone or in combination with inhaled therapy and inhaled therapy alone. Subsequently, acute exacerbations were compared between two groups during the 24-week follow-up period. FEF25%-75% in 43 patients with clinically controlled asthma given oral anti-inflammatory agents alone or in combination with inhaled therapy was significantly higher than that in 49 patients given inhaled therapy alone. Meanwhile, the percentage of eosinophils and levels of IL-5 and ECP in patients with clinically controlled asthma given oral anti-inflammatory agents alone or in combination with inhaled therapy were significantly lower than those in patients given inhaled therapy alone. Additionally, the patients with clinically controlled asthma given inhaled therapy were likely to have more acute exacerbation than the patients given oral anti-inflammatory agents alone or in combination with inhaled therapy during the 24-week follow-up period. Systemic anti-inflammatory agents may have a greater effect on parameters reflecting small airway patency and reducing acute exacerbations, presumably secondary to reduction in airway inflammation.

Performance of Eosinophil Cationic Protein as a Biomarker in Asthmatic Children

Although blood eosinophils are a frequently used marker of type 2 inflammation in children with asthma, their sensitivity is relatively poor. Additional markers of type 2 inflammation are needed. We hypothesized that plasma concentrations of eosinophil cationic protein (ECP), a marker of eosinophil activation, would be useful for detection of type 2 inflammation and would predict poorer asthma outcomes over 1 year. Children and adolescents 6 through 17 years (N= 256) with confirmed asthma completed a baseline visit and a follow-up visit at 12 months. A subsetalso underwent systemic corticosteroid responsiveness testing with intramuscular triamcinolone. Outcome measures at 12 months included uncontrolled asthma, lung function, and asthma exacerbations treated with systemic corticosteroids. Plasma ECP concentrations ranged from 0.03 to 413.61 ng/mL (median, 6.95 ng/mL) and were consistently associated with other markers of type 2 inflammation. At baseline, children in the highest ECP tertile had poorer asthma control, more airflow limitation, and more exacerbations, but also had greater symptom improvement with intramuscular triamcinolone. At 12 months, associations between the highest ECP tertile and exacerbations, but not lung function or asthma control, persisted after covariate adjustment. However, the sensitivity of ECP was modest and was not markedly different from that of blood eosinophil counts. Plasma ECP concentrations may be a useful marker of type 2 inflammation in children and may help identify those children at highest risk for recurrent exacerbations who could benefit from corticosteroid treatment. However, additional markers may be needed to improve sensitivity for outcome detection.