Sputum Cell Counts Research Articles

Changing the airway pH

Background Although airway hyperacidity may have a great effect in the pathogenesis of asthma, its evaluation may be difficult. This study was conducted at Mansoura University Hospitals aiming to evaluate the degree of asthma control by increasing pH in uncontrolled asthma cases by the use of inhaled NaHCO3 8.4% for 3 months. Patients and methods A total of 100 patients with asthma were included, and they were divided by simple randomization to 75 patients in study group and 25 controls. All participants were subjected to history taking, clinical examination, sputum cell count and pH assessment, PFT, high-resolution computed tomography, and sputum induction and NaHCO3 8.4% inhalation. Evaluation of asthma exacerbation 6 months before and after intervention was carried out. Results There was no significant difference between study and control groups regarding demographic data, all basic investigations, pulmonary function, or sputum total and differential cells. We found a significant increase of sputum pH after treatment in the study group. Furthermore, significant improvement of forced expiratory volume in 1 s, forced vital capacity, four parameters of control assessment, and exacerbation also was seen in the study group after treatment. There was a significant decrease of total cells and eosinophils in sputum in the study group. The linear regression model showed a significant negative correlation for all the studied factors except short-acting beta 2 (SABA) and day symptom improvement. Conclusion We conclude that the decrease of airway acidity by NaHCO3 8.4% leads to significant decrease in inflammatory cells and frequency of exacerbation and improves pulmonary function in poorly controlled bronchial asthma.

Treatment with the anti-IgE monoclonal antibody omalizumab in women with asthma undergoing fertility treatment: a proof-of-concept study—The PRO-ART study protocol

IntroductionAsthma is associated with prolonged time to pregnancy and a higher need for fertility treatment. However, the mechanism underlying this association remains incompletely understood. Previous research points to asthma-driven systemic...

Risk factors associated with frequent exacerbations in asthma

BackgroundAsthma is a chronic airway inflammatory disease with various degrees of severity. Exacerbations are commonly seen in uncontrolled asthma and their treatment involves oral corticosteroids use with a lot of side effects. ObjectiveThe aim of the study was to identify easily available predictors for future exacerbations in patients with asthma. MethodsThis is a prospective study on 250 consecutive patients with asthma with a successful sputum induction. Exacerbation rate in the following year was assessed by telephone interview. Logistic regression was used to test the relationship between the binary outcomes (<1 or ≥1 exacerbation, <2 or ≥2 exacerbations) and a set of covariates including demographic, clinical, functional and inflammatory characteristics such as FeNO, sputum and blood cell counts. The results were then applied and validated in a new cohort of 1450 patients. ResultsSputum and blood eosinophils were able to identify patients presenting ≥1 or ≥2 exacerbations with the same discriminative power (AUC:0.65 and 0.64 respectively). The multiple regression analysis identified that exacerbations in the previous year (OR = 9.3), treatment with high doses ICS (OR = 27.1), blood eosinophils (cells/mm³, OR = 1.8) and FEV1/FVC (OR = 0.93) were independent predictors of exacerbations in the year following the visit with an AUC of 0.93 for this model. Frequent exacerbations (≥2) were also predicted by exacerbations in the previous year (OR = 10.5), treatment with high doses ICS (OR = 39.2) and blood eosinophils (OR = 3.5) with an AUC of 0.95 for the model. ConclusionBlood and sputum eosinophils have similar predictive value for future exacerbations. Prediction could be improved by combining this information with lung function, ICS dose and history of previous exacerbations.

Sputum Inflammometry to Manage Chronic Obstructive Pulmonary Disease Exacerbations: Beyond Guidelines.

Quantitative sputum cytometry facilitates in assessing the nature of bronchitis associated with exacerbations of chronic obstructive pulmonary disease (COPD). This is not assessed in most clinical trials that evaluate the effectiveness of strategies to prevent or to treat exacerbations. While up to a quarter of exacerbations may be associated with raised eosinophil numbers, the vast majority of exacerbations are associated with neutrophilic bronchitis that may indicate airway infections. While eosinophilia may be a predictor of response to corticosteroids (oral and inhaled), the limited efficacy of anti–interleukin 5 therapies would suggest that eosinophils may not directly contribute to those exacerbations. However, they may contribute to airspace enlargement in patients with COPD through various mechanisms involving the interleukin 13 and matrix metalloprotease pathways. The absence of eosinophils may facilitate in limiting the unnecessary use of corticosteroids. The presence of neutrophiia could prompt an investigation for the specific pathogens in the airway. Additionally, sputum measurements may also provide insight into the mechanisms of susceptibility to airway infections. Iron within sputum macrophages, identified by hemosiderin staining (and by more direct quantification) may impair macrophage functions while the low levels of immunoglobulins in sputum may also contribute to airway infections. The assessment of sputum at the time of exacerbations thus would facilitate in customizing treatment and treat current exacerbations and reduce future risk of exacerbations.

DIFFERENTIATION OF ASTHMA, COPD, AND ASTHMA-COPD OVERLAP VIA A SIMPLIFIED SPUTUM CELL COUNT METHOD

DIFFERENTIATION OF ASTHMA, COPD, AND ASTHMA-COPD OVERLAP VIA A SIMPLIFIED SPUTUM CELL COUNT METHOD

Effect of the inhaled PDE4 inhibitor CHF6001 on biomarkers of inflammation in COPD

BackgroundCHF6001 is a novel inhaled phosphodiesterase-4 inhibitor. This Phase IIa study assessed the effects of CHF6001 on markers of inflammation in induced sputum and blood in patients with chronic obstructive pulmonary disease (COPD).MethodsThis was a multicentre, three-period (each 32 days), three-way, placebo-controlled, double-blind, complete-block crossover study. Eligible patients had COPD, chronic bronchitis, and were receiving inhaled triple therapy for ≥2 months. Patients received CHF6001 800 or 1600 μg, or matching placebo twice daily via multi-dose dry-powder inhaler (NEXThaler). Induced sputum was collected pre-dose on Day 1, and post-dose on Days 20, 26 and 32. Blood was sampled pre-dose on Day 1, and pre- and post-dose on Day 32.ResultsOf 61 randomised patients, 54 (88.5%) completed the study. There were no significant differences between groups for overall sputum cell count, or absolute numbers of neutrophils, eosinophils or lymphocytes. CHF6001 800 μg significantly decreased the absolute number and percentage of macrophages vs placebo.In sputum, compared with placebo both CHF6001 doses significantly decreased leukotriene B4, C-X-C motif chemokine ligand 8, macrophage inflammatory protein 1β, matrix metalloproteinase 9, and tumour necrosis factor α (TNFα). In blood, both CHF6001 doses significantly decreased serum surfactant protein D vs placebo. CHF6001 1600 μg significantly decreased TNFα ex-vivo (after incubation with lipopolysaccharide).ConclusionThe data from this study show that CHF6001 inhaled twice daily has anti-inflammatory effects in the lungs of patients with COPD already treated with triple inhaled therapy.Trial registrationThe study is registered on ClinicalTrials.gov (NCT03004417).

Soluble fibre supplementation with and without a probiotic in adults with asthma: A 7-day randomised, double blind, three way cross-over trial

SummaryBackgroundSoluble fibre modulates airway inflammation in animal models. The aim of this study was to investigate the effects of soluble fibre supplementation, with and without a probiotic, on plasma short chain fatty acids (SCFA), airway inflammation, asthma control and gut microbiome in adults with asthma.MethodsA randomised, double-blinded, placebo controlled 3-way cross-over trial in 17 subjects with stable asthma at the Hunter Medical Research Institute, Newcastle, Australia. Subjects received 3 × 7 day oral interventions in random order; soluble fibre (inulin 12 g/day), soluble fibre + probiotic (inulin 12 g/day + multi-strain probiotic >25 billion CFU) and placebo. Plasma SCFA, sputum cell counts and inflammatory gene expression, asthma control gut microbiota, adverse events including gastrointestinal symptoms were measured.FindingsThere was no difference in change in total plasma SCFA levels (μmol/L) in the placebo versus soluble fibre (Δmedian [95% CI] 16·3 [−16·9, 49·5], p = 0·335) or soluble fibre+probiotic (18·7 [−14·5, 51·9], p = 0·325) group. Following the soluble fibre intervention there was an improvement in the asthma control questionnaire (ACQ6) (∆median (IQR) -0·35 (−0·5, −0·13), p = 0·006), sputum %eosinophils decreased (−1.0 (−2·5, 0), p = 0·006) and sputum histone deacetylase 9 (HDAC9) gene expression decreased (−0.49 (−0.83, −0.27) 2-ΔCt, p = .008). Individual bacterial operational taxonomic units changed following both inulin and inulin+probiotic arms.InterpretationSoluble fibre supplementation for 7 days in adults with asthma did not change SCFA levels. Within group analysis showed improvements in airway inflammation, asthma control and gut microbiome composition following inulin supplementation and these changes warrant further investigation, in order to evaluate the potential of soluble fibre as a non-pharmacological addition to asthma management.FundJohn Hunter Hospital Charitable Trust.

Assessment of breath volatile organic compounds in acute cardiorespiratory breathlessness: a protocol describing a prospective real-world observational study

IntroductionPatients presenting with acute undifferentiated breathlessness are commonly encountered in admissions units across the UK. Existing blood biomarkers have clinical utility in distinguishing patients with single organ pathologies but have...

Clinical and inflammatory phenotypic features of asthmatic patients sensitive to cold stimulation

To explore the clinical symptoms, lung function and airway inflammation phenotype characteristics of asthmatic patients who are sensitive to cold stimulation. Eighty patients with newly diagnosed bronchial asthma or with mild to moderate acute exacerbation of previously diagnosed bronchial asthma but without regular treatment were selected. According to whether cold air stimulation could induce respiratory symptoms such as cough and wheeze, the patients were divided into cold-insensitive group (45 cases) and cold-sensitive group (35 cases). All the patients were treated with inhaled corticosteroid (ICS), long-acting β2 receptor agonist (LABA; salmeterol xinafoate and fluticasone propionate powder for inhalation, 50 μg/250 μg, twice daily) and montelukast sodium tablets (10 mg, once daily); short-acting β2 receptor agonist (SABA) and/or systemic glucocorticoid (prednisone acetate tablets, 10 mg, once daily; or injection of methylprednisolone sodium succinate, 40 mg) were given if necessary. Asthma Control Test (ACT) score before treatment and at 3 months of treatment was used to assess the clinical symptoms such as cough and wheeze; spirometry was performed to determine lung function impairment and recovery. Blood and induced sputum cell counts were examined to determine the characteristics of airway inflammation. The two groups were comparable for age, gender, BMI, proportion of smokers and allergic rhinitis before treatment. The cold-sensitive patients experienced significantly more frequent acute exacerbations than the cold-insensitive patient within 1 year before the visit (P &lt; 0.05), but the use of SABA and glucocorticoid for symptom control during the treatment did not differ significantly between the two groups (P &gt; 0.05). The ACT scores of the cold-sensitive group were significantly lower than those of the cold-insensitive group both before and after the treatment (P &lt; 0.01). Compared with the cold-insensitive patients, the cold-sensitive patients had more obvious impairment of FEV1/FVC% and FEV1%pred before treatment (P &lt; 0.01), and also showed poorer recovery after treatment (P &lt; 0.05). The percentages of eosinophils in blood and induced sputum samples did not differ significantly between the two groups either before and after the treatment, but the percentage of neutrophils was significantly higher in the cold-sensitive group (P &lt; 0.01). In the induced sputum samples collected before treatment, the cell populations consisted mainly of eosinophilic subtype (60%) and neutrophilic subtype (20%) in the cold-insensitive group; in the cold-sensitive patients, the sputum neutrophilic subtype cells increased significantly to 42.86% (P=0.03) and the eosinophilic subtype cells were lowered to 31.43% (P=0.01). The cold-sensitive asthmatic patients experience frequent recurrent and/or aggravated symptoms and have obvious lung function impairment. Different from that in patients with classic asthma, the airway inflammatory phenotype in these patients is characterized by the domination by neutrophilic subtype.

Optimizing sputum cell counts prior to bronchial thermoplasty: A preliminary report

RATIONALE: There is considerable variability in patient outcomes following bronchial thermoplasty (BT), an adjunct therapy for patients with severe asthma. It is unclear if the presence of airway inflammation is important to consider prior to BT.OBJECTIVE: The objective of the article was to ascertain if optimizing airway inflammation guided by sputum cell counts prior to BT, as compared to guideline-based-care, results in better patient outcomes and improved airway hyperresponsiveness.METHODS: We retrospectively evaluated asthmatics who underwent BT at our institution (Firestone Institute for Respiratory Health, St. Joseph’s Healthcare Hamilton, Canada) and compared clinical outcomes (asthma-quality-of-life-questionnaire [AQLQ], asthma-control-questionnaire [ACQ] and provocative concentration of methacholine required to decrease the forced expiratory volume in one second by 20% [PC20]) in those patients managed using guideline-based-care (n = 7) versus a protocol that involved optimizing airway inflammation using quantitative sputum cell counts (n = 7).MEASUREMENTS AND MAIN RESULTS: Following BT, there were greater improvements in the inflammation-optimized group than in the guideline-based-care group for the asthma-quality-of-life-questionnaire (ΔAQLQ: 2.2 ± 1.2 vs 0.8 ± 0.9, p = 0.04), asthma-control-questionnaire (ΔACQ: −2.3 ± 1.3 vs −0.5 ± 1.1, p = 0.02) and airway hyperresponsiveness (ΔPC20: 3.9 ± 1.2 vs. 0.6 ± 0.8 doubling-doses, p = 0.008).CONCLUSION: This is the first study to provide preliminary evidence that BT, when performed after normalizing sputum cell counts, significantly improves patient reported outcomes and airway hyperresponsiveness, compared to current guidelines that presume airway inflammation is controlled.

Depressive symptom‐associated IL‐1β and TNF‐α release correlates with impaired bronchodilator response and neutrophilic airway inflammation in asthma

Depressive symptoms worsen asthma outcomes; however, the mechanism remains largely unexplored. This study aimed to determine whether depressive symptom-associated immune inflammation correlates with impaired bronchodilator response (BDR) and airway inflammatory phenotypes. Eligible adults with asthma (n=198) underwent clinical assessment, sputum induction and blood sampling. Depressive symptoms were defined by scores on the depression subscale of the Hospital Anxiety and Depression Scale (HADS-D). Pre- and post-bronchodilator spirometry was performed for BDR. Airway inflammatory phenotypes were defined by sputum cell counts. CRP, IL-1β, IL-5, IL-6, IL-8, TNF-α, IFN-γ, CCL17 and CCL22 in serum and sputum were detected. Compared with the non-depressive group (n=174), the depressive group (n=24) exhibited impaired BDR (P=0.032) and increased sputum neutrophils (P=0.023), which correlated with the HADS-D scores (P=0.027 and P=0.029). Levels of IL-1β, TNF-α and IFN-γ in the serum and those of IL-1β and IFN-γ in the sputum were elevated in the depressive group compared to those in the non-depressive group (all P<0.05). Multiple regression models indicated that TNF-α in the sputum and IL-1β, IL-6 and IFN-γ in both the serum and sputum were inversely associated with BDR; TNF-α in the sputum and IL-1β in both the serum and sputum were positively correlated with sputum neutrophils. Mediation analyses revealed that IL-1β and TNF-α in the sputum and IL-1β in both the serum and sputum mediate the correlations of the HADS-D scores with BDR and sputum neutrophils, respectively. Asthma patients with depressive symptoms present worse asthma control, which is most likely explained by impaired BDR and neutrophilic airway inflammation. IL-1β and TNF-α, which are two key pro-inflammatory cytokines that mediate the correlation of depressive symptoms with impaired BDR and neutrophilic airway inflammation, may serve as targeted biomarkers in the neuropsychological phenotype of asthma; however, this result needs to be further validated.

Occupational asthma: clinical phenotypes, biomarkers, and management.

This review focuses on new findings in the clinical and inflammatory aspects that can help to better identify the different phenotypes of work-related asthma and the development of specific biomarkers useful in diagnosis and follow-up. Studies on phenotyping of occupational asthma, a subtype of work-related asthma, have mainly compared the clinical, physiological, and inflammatory patterns associated with the type of agent causing occupational asthma, namely, high-molecular-weight and low-molecular-weight agents. Most of this research has found that patients with occupational asthma due to high-molecular-weight agents have an associated presence of rhinitis, conjunctivitis, atopy, and a pattern of early asthmatic reactions during specific inhalation challenge. The inflammatory profile (blood eosinophils, sputum cell count, or exhaled nitric oxide) may be similar when occupational asthma is caused by either type of agent. In some studies, severity of asthma and exacerbations have been associated with exposure to low-molecular-weight agents. The most reliable biomarkers in diagnosis and follow-up are eosinophilia in induced sputum and exhaled nitric oxide. There are several phenotypes, characterized by its pathogenesis and inflammatory profile. Avoidance of the causative agents does not warrant complete recovery of occupational asthma. Treatment with biologic agents may be considered in severe occupational asthma.

Paucigranulocytic asthma: Uncoupling of airway obstruction from inflammation

Among patients with asthma, heterogeneity exists regarding the pattern of airway inflammation and response to treatment, prompting the necessity of recognizing specific phenotypes. Based on the analysis of inflammatory cell counts in induced sputum, asthmatic patients can be classified into 4 unique phenotypes: eosinophilic asthma, neutrophilic asthma, mixed granulocytic asthma, and paucigranulocytic asthma (PGA). PGA is an asthma phenotype with no evidence of increased numbers of eosinophils or neutrophils in sputum or blood and in which anti-inflammatory therapies are ineffective at controlling symptoms. Although underinvestigated, PGA is the most common asthma phenotype in patients with stable asthma. However, PGA is sometimes underestimated because of the exclusive reliance on induced sputum cell counts, which are variable among cohorts of studies, prompting the necessity of developing improved biomarkers. Importantly, investigators have reported that inhaled corticosteroids had a limited effect on airway inflammatory markers in patients with PGA and therefore defining PGA as a potentially "steroid-insensitive" phenotype that requires exploration of alternative therapies. PGA manifests as an uncoupling of airway obstruction from airway inflammation that can be driven by structural changes within the airways, such as airway smooth muscle tissue hypertrophy. Animal models provide evidence that processes evoking airway hyperresponsiveness and airway smooth muscle thickening occur independent from inflammation and might be a consequence of a loss of negative homeostatic processes. Collectively, further understanding of PGA with a focus on the characterization, prevalence, clinical significance, and pathobiology derived from animal studies will likely provideprecision therapies that will improve PGA clinical outcomes.

Diagnostic value of fractional exhaled nitric oxide for asthma-chronic obstructive pulmonary disease overlap syndrome.

To examine the difference in the fractional exhaled nitric oxide (FeNO) between chronic obstructive pulmonary disease (COPD) patients with asthma-COPD overlap syndrome (ACOS) and patients with Non-ACOS COPD (Non-ACOS) and to investigate the correlation between FeNO levels and the differential cell counts of eosinophils in induced sputum, in order to explore the diagnostic value of FeNO in ACOS.A prospective, case-control study was performed on 53 cases of ACOS group and 53 cases of Non-ACOS group in the Respiratory Medicine Outpatient of Zhangzhou Municipal TCM Hospital, Affiliated to Fujian University of Traditional Chinese Medicine. The FeNO levels and induced sputum cell counts were determined and the correlation between FeNO levels and eosinophile percentage was analyzed by Pearson linear correlation analysis.The FeNO levels in patients with ACOS (37[24.5–53.0]) ppb were significantly higher than those of patients with Non-ACOS (20 [15.5–24.5] ppb) (P < .01). Also, the percentage of eosinophils in induced sputum in the ACOS group (5.70 [1.50–17.62]%) were significantly higher than those of the Non-ACOS group (0.50 [0.00–1.00]%) (P < .01). FeNO in both groups correlated positively with the percentage of eosinophils in induced sputum (P < .01), with a correlation coefficient r of 0.521. The area under the receiver operating curve of FeNO for the diagnosis of ACOS phenotype was 0.815 (P < .01), the sensitivity and specificity reach highest when the cut off value was 25.50 ppb.The FeNO in patients from the ACOS group were significantly higher than those in Non-ACOS group and were moderately correlated with the percentage of eosinophils in induced sputum. The results indicated that FeNO may be used as a diagnostic index for ACOS, in addition to the induced sputum.

Predictive value of eosinophils and neutrophils on clinical effects of ICS in COPD.

Inflammation is present to a variable degree and composition in patients with COPD. This study investigates associations between both eosinophils and neutrophils in blood, sputum, airway wall biopsies and bronchoalveolar lavage (BAL) and their potential use as biomarkers for clinical response to inhaled corticosteroids (ICS). In total, 114 steroid-naïve COPD patients of the Groningen Leiden Universities Corticosteroids in Obstructive Lung Disease (GLUCOLD) study using ICS or placebo during 30-month follow-up were included. Cell counts in blood, sputum, biopsies and BAL were evaluated at baseline. In addition, at baseline, 6 and 30 months, forced expiratory flow in 1 s (FEV1 ), residual volume/total lung capacity (hyperinflation) and Clinical COPD Questionnaire were evaluated. Cross-sectional analyses at baseline showed that higher blood eosinophils were significantly associated with higher eosinophil counts in sputum, biopsies and BAL. However, blood neutrophils did not significantly correlate with neutrophil counts in the other compartments. Baseline eosinophils and neutrophils, in whichever compartment measured, did not predict longitudinal FEV1 changes. Higher baseline biopsy eosinophils were associated with an increase in symptoms during 6- and 30-month ICS treatment. In addition, higher biopsy neutrophils were associated with a more marked reduction in hyperinflation during 6-month ICS treatment compared with placebo. Our findings indicate that blood eosinophils reflect eosinophils in other compartments, in contrast to neutrophils, in ICS-naïve COPD patients. Both baseline eosinophils and neutrophils do not predict ICS-induced lung function changes over a period of 6-30 months. The associations of biopsy eosinophils with worsening respiratory symptoms and biopsy neutrophils with improvement in hyperinflation during ICS treatment deserve further investigation.

Breathomics for Assessing the Effects of Treatment and Withdrawal With Inhaled Beclomethasone/Formoterol in Patients With COPD.

Background: Prospective pharmacological studies on breathomics profiles in COPD patients have not been previously reported. We assessed the effects of treatment and withdrawal of an extrafine inhaled corticosteroid (ICS)-long-acting β2-agonist (LABA) fixed dose combination (FDC) using a multidimensional classification model including breathomics.Methods: A pilot, proof-of-concept, pharmacological study was undertaken in 14 COPD patients on maintenance treatment with inhaled fluticasone propionate/salmeterol (500/50 μg b.i.d.) for at least 8 weeks (visit 1). Patients received 2-week treatment with inhaled beclomethasone dipropionate/formoterol (100/6 μg b.i.d.) (visit 2), 4-week treatment with formoterol alone (6 μg b.i.d.) (visit 3), and 4-week treatment with beclomethasone/formoterol (100/6 μg b.i.d.) (visit 4). Exhaled breath analysis with two e-noses, based on different technologies, and exhaled breath condensate (EBC) NMR-based metabolomics were performed. Sputum cell counts, sputum supernatant and EBC prostaglandin E2 (PGE2) and 15-F2t-isoprostane, fraction of exhaled nitric oxide, and spirometry were measured.Results: Compared with formoterol alone, EBC acetate and sputum PGE2, reflecting airway inflammation, were reduced after 4-week beclomethasone/formoterol. Three independent breathomics techniques showed that extrafine beclomethasone/formoterol short-term treatment was associated with different breathprints compared with regular fluticasone propionate/salmeterol. Either ICS/LABA FDC vs. formoterol alone was associated with increased pre-bronchodilator FEF25−75% and FEV1/FVC (P = 0.008–0.029). The multidimensional model distinguished fluticasone propionate/salmeterol vs. beclomethasone/formoterol, fluticasone propionate/salmeterol vs. formoterol, and formoterol vs. beclomethasone/formoterol (accuracy > 70%, P < 0.01).Conclusions: Breathomics could be used for assessing ICS treatment and withdrawal in COPD patients. Large, controlled, prospective pharmacological trials are required to clarify the biological implications of breathomics changes. EUDRACT number: 2012-001749-42.

Sputum proteomics and airway cell transcripts of current and ex-smokers with severe asthma in U-BIOPRED: an exploratory analysis.

Severe asthma patients with a significant smoking history have airflow obstruction with reported neutrophilia. We hypothesise that multi-omic analysis will enable the definition of smoking and ex-smoking severe asthma molecular phenotypes.The U-BIOPRED cohort of severe asthma patients, containing current-smokers (CSA), ex-smokers (ESA), nonsmokers and healthy nonsmokers was examined. Blood and sputum cell counts, fractional exhaled nitric oxide and spirometry were obtained. Exploratory proteomic analysis of sputum supernatants and transcriptomic analysis of bronchial brushings, biopsies and sputum cells was performed.Colony-stimulating factor (CSF)2 protein levels were increased in CSA sputum supernatants, with azurocidin 1, neutrophil elastase and CXCL8 upregulated in ESA. Phagocytosis and innate immune pathways were associated with neutrophilic inflammation in ESA. Gene set variation analysis of bronchial epithelial cell transcriptome from CSA showed enrichment of xenobiotic metabolism, oxidative stress and endoplasmic reticulum stress compared to other groups. CXCL5 and matrix metallopeptidase 12 genes were upregulated in ESA and the epithelial protective genes, mucin 2 and cystatin SN, were downregulated.Despite little difference in clinical characteristics, CSA were distinguishable from ESA subjects at the sputum proteomic level, with CSA patients having increased CSF2 expression and ESA patients showing sustained loss of epithelial barrier processes.

Plasma miR-199a-5p is increased in neutrophilic phenotype asthma patients and negatively correlated with pulmonary function.

ObjectiveWe investigated the relationship between plasma miRNAs levels and inflammatory characteristics in asthmatic patients.MethodsEligible adults with untreated asthma (n = 35) underwent a clinical assessment, sputum induction, and assessment of pulmonary function test and Asthma Control Test (ACT) scores. Asthma phenotypes were defined using the sputum cell count. miR-199a-5p expression was measured using quantitative real-time polymerase chain reaction (qPCR). Lipopolysaccharide (LPS) stimulation was used to detect miR-199a-5p secretion from peripheral blood-derived neutrophil, lymphocyte, macrophage and BEAS-2B cells. The correlation of miR-199a-5p expression with clinical parameters was analyzed using multiple linear regression analysis. In silico analysis predicted the target genes and signaling pathway of miR-199a-5p. Transfection of miR-199a-5p mimics in human airway smooth muscle cells (HASMCs) was performed in vitro.ResultsThe miRNA-199a-5p levels in plasma and sputum increased significantly in patients with neutrophilic asthma compared to healthy subjects (ps = 0.014 and 0.006, respectively). Expression of miR-199a-5p in the plasma of asthmatic patients positively correlated with sputum miR-199a-5p expression (r = 0.511, p = 0.021). The miR-199a-5p level was only elevated with LPS stimulation in neutrophils but not macrophages, lymphocytes, or epithelial cells from healthy controls (p < 0.01). miR-199a-5p expression increased in response to LPS (p = 0.005) and LPS combined with IL-4 (p = 0.003), but not IL-4 alone. However, peripheral neutrophils from eosinophilic asthma patients did not respond to LPS with increased miR-199a-5p expression (n = 5, p > 0.05) in contrast to the significant response from neutrophilic patients (n = 4, p < 0.0001). miR-199a-5p negatively correlated with FEV1, FVC and PEF (r = -0.377, p = 0.026; r = -0.419, p = 0.012; and r = -0.392, p = 0.024, respectively). Multivariate correlation analysis confirmed that the plasma miR-199a-5p levels negatively correlated with FEV1 in patients with asthma (Adjusted R2 = 0.164, p = 0.015). In silico analysis suggested that the WNT signaling pathway participates in miR-199a-5p mediation of smooth muscle cell hypertrophy. In vitro experiment, miR-199a-5p mimics inhibited the protein expressions of WNT2 and WNT4, decreased the c-myc expression and dramatically increased the Sm-MHC expression in HASMCs.ConclusionPlasma miR-199a-5p was increased in neutrophilic asthma and negatively correlated with pulmonary function, which suggests that miR-199a-5p actively contributes to disease pathogenesis by modulating the inflammatory process and transferring the signal from inflammatory cells to structure cells.

Asthma Phenotypes Defined From Parameters Obtained During Recovery From a Hospital-Treated Exacerbation

Asthma is a heterogeneous disease with diverse clinical manifestations and inflammatory pathologies that is punctuated by exacerbations. To describe the clinical and inflammatory characteristics of patients with asthma treated in hospital for an acute exacerbation. Data from 320 adult patients receiving treatment for an acute exacerbation of asthma were obtained. In 218 patients with complete data, we used the Ward hierarchical clustering to obtain clusters. Pulmonary function, blood cell counts, sputum cell counts, serum IgE levels, and fractional exhaled nitric oxide were measured on hospital admission. We selected 13 variables with which we performed the Ward minimum-variance hierarchical clustering. Four clusters were defined. Clusters 1 (24.5%) and 3 (36.7%) were characterized by predominantly female patients with asthma with sputum neutrophilia, with cluster 1 associated with a small degree of airflow obstruction and early-onset asthma and cluster 3 with a moderate degree of reduction in FEV1. Clusters 2 (22.0%) and 4 (16.5%) were associated with high sputum eosinophilia and severe airflow obstruction. Cluster 4 was made exclusively of male smoking subjects, whereas cluster 2 was made up of predominantly female nonsmoking subjects with the worst FEV1, forced expiratory flow at 25% to 75% of forced vital capacity (% predicted), and partial pressure of oxygen in arterial blood on admission. There were no differences between clusters in terms of atopy, serum IgE, prevalence of nasal disease, dose of maintenance inhaled corticosteroids, or oral/systemic corticosteroid use and asthma exacerbations. The clusters during recovery from an exacerbation of asthma were distinguished by airflow obstruction and a neutrophilic, eosinophilic, or mixed inflammation. Eosinophilic inflammation was found in smoking and nonsmoking patients with asthma during an exacerbation.

Blood eosinophilia in chronic obstructive pulmonary disease: is there a relation with airway eosinophilia?

Background Blood eosinophilia was considered an indicator of sputum eosinophilia in asthmatic patients. Nevertheless, its role in chronic obstructive pulmonary disease (COPD) was not completely examined. Aim The aim was to investigate if peripheral blood eosinophilic cells represent an indicator (marker) of eosinophilia of the airway in stable cases. Patients and methods The trial was held at Al-Azhar University Hospital (Damietta), between October 2015 and March 2017. It included 264 persons with stable COPD. All cases were assessed for demographic features, smoking history, history of exacerbation in the last year, dyspnea, associated comorbidities, quality of life, lung functions, sputum, and peripheral blood eosinophilic cell counts. The included participants were allocated into two sets: group with eosinophilia (n=76) and patients without eosinophilia (n=188). Results There was proportional correlation between eosinophilia of the blood and sputum; and blood eosinophilia can differentiate between cases who had or had no sputum eosinophilia at a cutoff of 2.5%. In addition, eosinophil/lymphocyte ratio and eosinophil/neutrophil ratio were higher in eosinophilic COPD and are associated ELR with eosinophilia of the sputum ENR. Conclusion There is correlation between eosinophils of the peripheral blood and sputum in stable cases. Thus, the count of blood eosinophil may act as a dependable marker for eosinophilic COPD