The expression of IL17RA on sputum macrophages in asthma patients.

IL-17A and IL-25 (IL-17 cytokines family) play an important role in the development of asthma, and allergy. Both cytokines act by binding to heterodimeric receptors with IL17RA as a common subunit. This receptor is found on macrophages, and some other cell types. The aim of the study was to determine the expression of IL17RA on asthmatic and control macrophages from induced sputum (IS) with the regard to IL-17/IL-25 background and relation to clinical features of the disease. We found an elevated expression of IL17RA on sputum macrophages in asthma patients vs controls. A characteristic sputum profile of atopic asthmatic was as follows: high CD206 + IL17RA + macrophage percentage, elevated IL-25 level and low CD206 + IL17RA- macrophage percentage. Based on the above results, it seems that CD206 + sputum macrophages are the effector cells that express common subunit of the receptor for IL-17A and IL-25 in asthma. This may be related to the Th2-dependent environment in asthma and increased concentrations of IL-25 and IL-13 as well as eosinophils in the airways. To our knowledge, our study provides the first data on a possible link between immunological reaction orchestrating CD206 + expressing sputum macrophages and IL-25 via IL17RA pathway in the asthmatic airways.

Expression of TSLP and IL-33 receptors on sputum macrophages of asthma patients and healthy subjects.

Objective: Local cytokine milieu (especially Th2 inflammatory type) secreted into the asthmatic airways affect the alternative activated macrophages polarization (M2). TSLP and IL-33 are important alarmins of allergic response associated with Th2 inflammation. The aim of the study was to investigate the expression of the receptors for epithelial derived cytokines: TSLP (TSLPR) and IL-33 (ST2) on induced sputum CD206 positive macrophages from asthma and healthy subjects and analyze the relationships between these receptors and clinical features of the disease. Methods: Immunofluorescence staining for CD206 and TSLPR or ST2 on sputum macrophages was performed in 20 adult patients with stable asthma - 75% with atopy (3 intermittent, 12 mild-to-moderate, 5 severe, of which 11 were on biological anty-IgE treatment) and 23 healthy adult controls - 48% with atopy. Results: Our study demonstrated an increased expression of TSLP and IL-33 receptors on bronchial CD206 positive macrophages in asthma group. TSLPR but not ST2 had also greater expression on CD206 negative macrophages in asthma patients. Increased expression of both investigated receptors was related to longer disease duration and impaired lung function. We observed increased count of CD206lowTSLPhigh macrophages as well as positive correlation of these cells with total serum IgE in patients with atopy. Conclusions: The macrophage response during allergic reaction is likely to be connected with TSLP but rather not with IL-33 action. Our study indicates an important role of crosstalk between macrophages, TSLP and IL-33 in asthma pathophysiology.

Impact of a Single Session of Inhalation Technique Training on Inhalation Skills and the Course of Asthma and COPD.

BACKGROUND: A significant percentage of patients with asthma and COPD do not use their inhalers properly. The aim of this study was to evaluate the impact of a single session of inhalation technique training on the frequency of inhalation errors and the course of asthma and COPD. METHODS: This randomized controlled trial included adults who had been diagnosed with and treated for asthma or COPD with at least one inhaler use daily on a regular basis. All subjects were followed for 6 months, at which time their inhalation technique was assessed and an individual inhalation training (study group) or a sham procedure (control group) was applied. The number of mishandlings was calculated as the ratio of the number of errors to the number of inhalers used by an individual subject. The effect of training was evaluated after 3 months and again after 6 months. RESULTS: 50 subjects with asthma and 50 subjects with COPD were enrolled. Only 20% of subjects made no critical errors before the intervention. Subjects who were trained in the proper inhalation technique made fewer errors after 3 months (32 of 50 vs 20 of 50). The relative risk was 1.63 (95% CI 1.1-2.4, P = .01) and the number needed to treat was 3.9 (95% CI 2.2-15). Despite the improvement in the inhalation technique, we found no reduction in the number of asthma/COPD exacerbations, symptom severity, or the quality of life. After 3 consecutive months, the efficacy of the intervention decreased, and only 66% of the former responders maintained the lower ratio of errors per inhaler. After 6 months, there was no difference in the number of subjects with better inhalation technique between intervention (24 of 50) and control group (27 of 50) (P = .62). CONCLUSIONS: Although a single inhalation training leads to a reduction in the number of errors made during inhalation, it does not influence the course of asthma and COPD. The positive effect of a single inhalation technique training is temporary. (ClinicalTrials.gov registration NCT02131454.).

Sputum interleukin-25 correlates with asthma severity: a preliminary study.

INTRODUCTION: Interleukin 25 is an epithelial-derived cytokine associated with allergic Th2 inflammation. However, little is known about the role of IL-25 in different asthma phenotypes and its relationship with disease severity. AIM: To evaluate and compare the mRNA and protein expression of IL-25 in patients with mild-to moderate/severe asthma and cough variant asthma (CVA). MATERIAL AND METHODS: Thirty-eight patients with stable asthma (11 patients with mild-to-moderate asthma, 14 patients with severe asthma and 13 patients with CVA) and 14 control subjects were enrolled. IL-25 protein concentration was measured in induced sputum (IS) supernatants by ELISA and IL-25 mRNA expression was evaluated in IS cells by real time PCR. RESULTS: No differences in IS IL-25 mRNA and IL-25 concentration between controls and the whole asthma group were found. In the detailed analysis, a lower IL-25 mRNA expression in sputum cells was observed in severe asthma compared to CVA and controls. IL-25 protein concentration in sputum supernatants was elevated in patients with severe asthma compared to controls, CVA and mild-to-moderate asthma. A sputum IL-25 level was increased in atopic vs. non-atopic asthma patients. The elevated IL-25 mRNA expression and protein concentration was associated with a lower eosinophil and higher neutrophil percentage in asthmatic airways. CONCLUSIONS: Our results suggest that IL-25 is particularly associated with severe asthma. The relationship between IL-25 and neutrophilic airway inflammation suggests the pleiotropic role of IL-25 in the immune response in this disease.

Expression of Inflammatory Mediators in Induced Sputum: Comparative Study in Asthma and COPD.

Asthma and COPD are the most common obstructive lung diseases characterized by inflammation in the lower airways which contribute to airflow limitation. Different inflammatory mediators are thought to play a key role in these diseases. This study was conducted in 13 patients with asthma, 12 patients with COPD, and 13 control subjects. The expression of mRNA of IL-6, IL-13, CXCL8, TSLP, IL-33, IL-25, IL-17, ECP, mast cell tryptase, CCL24, and CCL26 was assessed in induced sputum cells by real time PCR. We found that CXCL8 was strongly related to the neutrophil percentage but differed significantly in COPD and asthma patients. The expression of IL-17 was lower in patients with atopic asthma compared to non-atopic asthma. The percentage of macrophages correlated negatively with the expression of mast cell tryptase and ECP in COPD, and with CXCL8 in asthma. The expression of ECP correlated negatively with the severity of COPD symptoms measured by CAT. We conclude that asthma and COPD demonstrate a significant overlap in the airway cytokine profile. Thus, differentiation between the two diseases is difficult as based on a single cytokine, which suggests the coexistence of phenotypes sharing a common cytokine network in these obstructive lung diseases.

Expression of eotaxins in the material from nasal brushing in asthma, allergic rhinitis and COPD patients.

BACKGROUND: Asthma and COPD are non-infectious inflammatory diseases of the respiratory tract. Allergic rhinitis can be assumed as an intermediate condition between healthy and asthmatic state. Eotaxins are important indicators of allergic reaction. They are strong chemoattractants mainly for eosinophils but also for other cells. OBJECTIVE: We measured the level of eotaxin expression and inflammatory cell count in the material from nasal brushing in healthy controls and in patients with allergic rhinitis, asthma, and COPD. We studied the correlation between the eotaxin gene expression level in the material from nasal brushing and respiratory tests in asthma and COPD patients. METHODS: Expression of eotaxins was measured using quantitative RT-PCR. Number of eotaxin transcript copies was evaluated using real time PCR standard curve method. RESULTS: Of all eotaxins CCL24 had the highest expression in the material from nasal brushing, and its level was increased in allergic asthma. CCL11 was significantly increased in the material from nasal brushing of COPD patients. Increased levels of all three eotaxins were observed in the material from nasal brushing of patients with allergic rhinitis in season. The levels of CCL26 expression and FEV1/FVC factor were correlated negatively in the asthma group and positively in the COPD group. CONCLUSIONS: Eotaxins are crucial factors of allergic, asthmatic and also COPD inflammatory reactions. Our results suggest a dual role of CCL26 - it can act as a negative regulator for neutrophils in COPD, while in asthma it may act as a chemoatractant of eosinophils and other cells into the lung.

[Factors influencing asthma course and the degree of control in the patients assessed with own questionnaire and Asthma Control Test (ACT)]. / Wplyw wybranych czynników na przebieg i kontrole astmyu chorych poddanych badaniu za pomoca ankiety wlasneji Testu Kontroli Astmy (ACT).

INTRODUCTION: Asthma treatment requires control and monitoring. According to Global Initiative For Asthma (GINA) asthma severity is described by degree of control. The aim of the present study was to compare self-patients' opinions about asthma control with Asthma Control Test (ACT) scores. Furthermore factors associated with asthma control were analyzed in the examined group of patients. Age, BMI, the length and the degree of asthma treatment, concomitant diseases, physical activity, exposition to the tobacco smoke, needs for education and self-control were assessed. MATERIAL AND METHODS: The study was based on a 36-point questionnaire that evaluated last two years of asthma duration, the reference data were obtained by objective evaluation with ACT. Fifty three patients, 37 women and 16 men, median age 54 years (24-80 years), from outpatients clinic were enrolled into the study. According to ACT score the patients (pts) were divided into 3 groups: 25 points - well controlled asthma (group 1), 20-24 points - partially controlled asthma (group 2) and less than 20 points - lack of asthma control (group 3). Kruskal-Wallis test and Chi-square test were used to compare the characteristics of subjects in different ACT groups. Pearson's test was used for assessment of correlation between different parameters. RESULTS: Twenty seven of 53 pts (51%) were treated with low dose of inhaled steroids and long acting beta-agonists (third degree of treatment according to GINA). During last two years the treatment was intensified in 37 pts (70%) due to exacerbation, and 19 of them (36%) were hospitalized in the course of exacerbation. Although 36 out of 53 pts (68%) claimed their asthma was fully controlled, ACT showed full control only in 5/53 (9%) of cases, partial control in 18/53 (34%), lack of control - in 30/53 (57%). Older age (p 〈 0.05) and longer duration of the disease (p 〈 0.01) were the factors significantly influencing lack of asthma control. A tendency towards worse asthma control was combined also with overweight and greater number of concomitant diseases (mainly coronary artery disease, diabetes and gastro-esophageal reflux). The presence of influenza vaccinations in medical history was associated with better asthma control (p = 0.05). CONCLUSIONS: A discrepancy between subjective assessment of asthma control and ACT score was observed in the examined group of patients. The frequency of asthma exacerbations (according to questionnaire) was describing the degree of asthma control more precisely than self-assessment. Older age and longer disease duration were combined with significantly worse asthma control. Better asthma control was combined with the presence of influenza vaccinations in medical history.

IL-6 and IL-13 in induced sputum of COPD and asthma patients: correlation with respiratory tests.

BACKGROUND: IL-6 is strongly implicated in the development of chronic obstructive pulmonary disease (COPD). IL-13 is the well-documented central mediator in allergic asthma. IL-6 is attributed to the proinflammatory activities in COPD as well as asthma. In COPD patients exacerbation is increased by serum IL-6. The association of IL-13 as well as IL-6 with the impaired respiratory function of asthma patients remains controversial. OBJECTIVES: The aim of this study was to compare the concentration of IL-6 and IL-13 in the induced sputum of asthma and COPD patients, and to assess the possible association of these cytokines with the impairment of lung function. METHODS: Twenty-six subjects with COPD and 18 subjects with asthma were enrolled in this study. IL-6 and IL-13 levels were measured in induced sputum by ELISA and correlated with the results of respiratory tests. RESULTS: The induced sputum of COPD patients had a significantly higher IL-6 level than the sputum of asthma subjects while no significant differences were found in the levels of IL-13. There was a statistically significant negative correlation between IL-6 level and FEV(1) or FEV(1)/FVC in asthma patients (r = -0.59 and -0.54, respectively) and a negative correlation that did not reach statistical significance between IL-6 level and FEV(1), FEV(1)% or FVC in COPD subjects (r = -0.30, -0.30 and -0.38, respectively). There was no relationship between concentrations of IL-13 and impaired respiratory function. CONCLUSIONS: Our results confirmed that IL-6, but not of IL-13, is associated with respiratory disorders in both asthma and COPD patients.

[Macrophage phenotype in induced sputum in asthma subjects]. / Fenotyp makrofagów w plwocinie indukowanej u chorych na astme.

Macrophages represent the most predominant immune effector cells in the alveolar spaces and conducting airways and are known to express activated phenotype. The study was aimed at assessing the differences in cellular profile and the expression of selected surface markers on sputum macrophages in asthma and healthy subjects. 17 healthy subjects (never smoked) and 10 mild asthma subjects treated with glucocorticosteroids were enrolled into the study. For macrophage phenotyping a immunocytochemistry method was used with commercially available antibodies anti: CD14, CD71, CD11b and CD54. The nonparametric Mann Whitney U test was applied for data comparison, p value <0.05 being regarded as significant. The total number of cells were increased in asthma patients 4.81+/-5.27x 10(6) /ml vs healthy 2.8+/-2.15 x 10(6)/ml and it was statistically significant. Statistically significant increase in the percentage of eosinophils was observed in mild asthma subjects. No differences were found between the proportion in macrophages and lymphocytes. The macrophage phenothype in induced sputum differed in both groups. The expression of CD11b was higher in asthma group and the difference was statistically significant. The proportion of macrophages with the expression of CD 14, CD 71 and CD54 was comparable in both groups. Macrophage phenotyping during glucocorticosteroid therapy is useful in the assessment of inflammatory process in asthma subject.

[The influence oc glycocorticoid therapy on sputum ECP concentration in symptomatic patients with bronchial asthma]. / Wplyw leczenia glikokortykosteroidami na stezenie ECP w plwocinie indukowanej u chorych na astme oskrzelowa w okresie zaostrzenia.

UNLABELLED: ECP released from the granules of activated eosinophils is regarded to be a marker of airway inflammation in asthma. The study was performed to compare the usefulness of measuring serum and sputum ECP for monitoring the asthma treatment. 29 subjects with mild to moderate asthma (mean age 41 +/- 17) were admitted in exacerbation (FEV1 55.54 +/- 87.49% N). 10 subjects with grass pollen asymptomatic asthma and 10 healthy subjects were also enrolled in the study. Patients with symptomatic asthma were ordered 30 mg prednisone for 2 weeks and they continued during next 2 weeks inhaled budesonide therapy. The concentrations of ECP (mcg/L) were determined by CAP-system (Pharmacia). The total eosinophil count and serum ECP in all subjects treated orally and next by inhaled GKS didn't differ statistically. The highest sputum ECP concentration was determined in exacerbation of asthma 84.5 +/- 78 mcg/L and statistically were reduced after 2-weeks of prednisone treatment 24.4 +/- 12.1 mcg/L (p = 0.05). In following 2 weeks of budesonide treatment sputum ECP concentration was statistically negligible in relation to previous treatment in spite of increasing tendency (50 +/- 61.3 mcg/L (p = 0.2394). In asymptomatic grass pollen asthma sputum ECP concentration was 19.7 +/- 9.4 mcg/L, higher than in controls 12 +/- 5.8 mcg/L (p = 0.04). There were a significant correlations between total eosinophil count and serum (r = 0.6396) and sputum ECP(r = 0.4683) in exacerbation. CONCLUSIONS: 1. In asthma exacerbation elevated sputum ECP concentration was observed. 2. In consequence of prednisone treatment the sputum ECP concentration was reduced. 3. Sputum ECP measurement is more accurate than serum ECP for monitoring the effectiveness of treatment. 4. Sputum ECP concentration is a sensitive parameter which discriminate asymptomatic patients with asthma from healthy subjects.