Sputum Differential Cell Counts Research Articles

Nebulised interferon beta-1a (SNG001) in the treatment of viral exacerbations of COPD

BackgroundRespiratory viral infections are major drivers of chronic obstructive pulmonary disease (COPD) exacerbations. Interferon-β is naturally produced in response to viral infection, limiting replication. This exploratory study aimed to demonstrate proof-of-mechanism, and evaluate the efficacy and safety of inhaled recombinant interferon-β1a (SNG001) in COPD. Part 1 assessed the effects of SNG001 on induced sputum antiviral interferon-stimulated gene expression, sputum differential cell count, and respiratory function. Part 2 compared SNG001 and placebo on clinical efficacy, sputum and serum biomarkers, and viral clearance.MethodsIn Part 1, patients (N = 13) with stable COPD were randomised 4:1 to SNG001 or placebo once-daily for three days. In Part 2, patients (N = 109) with worsening symptoms and a positive respiratory viral test were randomised 1:1 to SNG001 or placebo once-daily for 14 days in two Groups: A (no moderate exacerbation); B (moderate COPD exacerbation [i.e., acute worsening of respiratory symptoms treated with antibiotics and/or oral corticosteroids]).ResultsIn Part 1, SNG001 upregulated sputum interferon gene expression. In Part 2, there were minimal SNG001–placebo differences in the efficacy endpoints; however, whereas gene expression was initially upregulated by viral infection, then declined on placebo, levels were maintained with SNG001. Furthermore, the proportion of patients with detectable rhinovirus (the most common virus) on Day 7 was lower with SNG001. In Group B, serum C-reactive protein and the proportion of patients with purulent sputum increased with placebo (suggesting bacterial infection), but not with SNG001. The overall adverse event incidence was similar with both treatments.ConclusionsOverall, SNG001 was well-tolerated in patients with COPD, and upregulated lung antiviral defences to accelerate viral clearance. These findings warrant further investigation in a larger study.Trial registrationEU clinical trials register (2017-003679-75), 6 October 2017.

Proinflammatory pattern in the lower airways of non-asthmatic obese adolescents

BackgroundObesity is known to be a pro-inflammatory condition affecting multiple organs. Obesity as a systemic pro-inflammatory state, might be associated with bronchial inflammation in non-smoking adolescents with a BMI ≥ 30 kg/m2 without evidence of concomitant chronic diseases. Materials and methodsWe studied non-asthmatic obese patients (n = 20; median age 15.8 years; BMI 35.0 kg/m2) compared to age matched healthy control subjects (n = 20; median age 17.5 years; BMI 21.5 kg/m2). Induced sputum differential cell counts and sputum mRNA levels were assessed for all study subjects. Serum levels of CRP, IL-6, and IL-8 were measured. Further, IL-5, IL-6, IL-8, IL-13, IL-17, TNF-α, IFN-γ, and IP-10 protein levels were analyzed in induced sputum was. ResultsSerum CRP levels, sputum inflammatory cell load and sputum eosinophils differed significantly between obese and non-obese subjects, for sputum neutrophils, a correlation was shown with BMI ≥ 30 kg/m2. Differences were also observed for sputum mRNA expression of IL6, IL8, IL13, IL17, IL23, and IFN-γ, as well as the transcription factors T-bet, GATA3, and FoxP3. ConclusionsIncreased bronchial inflammation, triggered by systemic or local inflammatory effects of obesity itself, may account for the higher rates of airway disease in obese adolescents.

Airway Bacteria Quantification Using Polymerase Chain Reaction Combined with Neutrophil and Eosinophil Counts Identifies Distinct COPD Endotypes.

Background: Chronic obstructive pulmonary disease (COPD) inflammatory endotypes are associated with different airway microbiomes. We used quantitative polymerase chain reaction (qPCR) analysis of sputum samples to establish the bacterial load upper limit in healthy controls; these values determined the bacterial colonisation prevalence in a longitudinal COPD cohort. Bacteriology combined with sputum inflammatory cells counts were used to investigate COPD endotypes. Methods: Sixty COPD patients and 15 healthy non-smoking controls were recruited. Sputum was analysed by qPCR (for Haemophilus influenzae, Moraxella catarrhalis, Streptococcus pneumoniae and Psuedomonas aeruginosa) and sputum differential cell counts at baseline and 6 months. Results: At baseline and 6 months, 23.1% and 25.6% of COPD patients were colonised with H. influenzae, while colonisation with other bacterial species was less common, e.g., S. pneumoniae—1.9% and 5.1%, respectively. H. influenzae + ve patients had higher neutrophil counts at baseline (90.1% vs. 67.3%, p < 0.01), with similar results at 6 months. COPD patients with sputum eosinophil counts ≥3% at ≥1 visit rarely showed bacterial colonisation. Conclusions: The prevalence of H. influenzae colonisation was approximately 25%, with low colonisation for other bacterial species. H. influenzae colonisation was associated with sputum neutrophilia, while eosinophilic inflammation and H. influenzae colonisation rarely coexisted.

IL-1\u03b2 augments TGF-\u03b2 inducing epithelial-mesenchymal transition of epithelial cells and associates with poor pulmonary function improvement in neutrophilic asthmatics

BackgroundNeutrophilic asthmatics (NA) have less response to inhaled corticosteroids. We aimed to find out the predictor of treatment response in NA.MethodsAsthmatics (n = 115) and healthy controls (n = 28) underwent clinical assessment during 6-month follow-up with standardized therapy. Asthmatics were categorized by sputum differential cell count. The mRNA expressions were measured by RT-qPCR for sputum cytokines (IFN-γ, IL-1β, IL-27, FOXP3, IL-17A, and IL-5). The protein of IL-1β in sputum supernatant was detected by ELISA. Reticular basement membranes (RBM) were measured in the biopsy samples. The role and signaling pathways of IL-1β mediating the epithelial-mesenchymal transition (EMT) process were explored through A549 cells.ResultsNA had increased baseline sputum cell IL-1β expression compared to eosinophilic asthmatics (EA). After follow-up, NA had less improvement in FEV1 compared to EA. For all asthmatics, sputum IL-1β mRNA was positively correlated with protein expression. Sputum IL-1β mRNA and protein levels were negatively correlated to FEV1 improvement. After subgrouping, the correlation between IL-1β mRNA and FEV1 improvement was significant in NA but not in EA. Thickness of RBM in asthmatics was greater than that of healthy controls and positively correlated with neutrophil percentage in bronchoalveolar lavage fluid. In vitro experiments, the process of IL-1β augmenting TGF-β1-induced EMT cannot be abrogated by glucocorticoid or montelukast sodium, but can be reversed by MAPK inhibitors.ConclusionsIL-1β level in baseline sputum predicts the poor lung function improvement in NA. The potential mechanism may be related to IL-1β augmenting TGF-β1-induced steroid-resistant EMT through MAPK signaling pathways.Trial registration: This study was approved by the Ethics Committee of Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology (IRB ID: 20150406).

Mixed Sputum Granulocyte Longitudinal Impact on Lung Function in the Severe Asthma Research Program.

Rationale: Some reports indicate longitudinal variability in sputum differential cell counts, whereas others describe stability. Highly variable sputum eosinophil percentages are associated with greater lung function loss than persistently elevated eosinophil percentages, but elevated neutrophils are linked to more severe asthma.Objectives: To examine sputum granulocyte stability or variability longitudinally and associations with important clinical characteristics.Methods: The SARP III (Severe Asthma Research Program III) cohort underwent comprehensive phenotype characterization at baseline and annually over 3 years. Adult subjects with acceptable sputum levels were assigned to one of three longitudinal sputum groups: eosinophils predominantly <2%, eosinophils predominantly ≥2%, or highly variable eosinophil percentages (>2 SDs determined from independent, repeated baseline eosinophil percentages). Subjects were similarly assigned to one of three longitudinal neutrophil groups with a 50% cut point.Measurements and Main Results: The group with predominantly <2% sputum eosinophils had the highest lung function (prebronchodilator FEV1% predicted, P < 0.01; FEV1/FVC ratio, P < 0.001) at baseline and throughout 3 years compared with other eosinophil groups. Healthcare use did not differ, although the highly variable eosinophil group reported more asthma exacerbations at Year 3. Longitudinal neutrophil groups showed few differences. However, a combination of predominantly ≥2% eosinophil and ≥50% neutrophil groups resulted in the lowest prebronchodilator FEV1% predicted (P = 0.049) compared with the combination with predominantly <2% eosinophils and<50% neutrophils.Conclusions: Subjects with predominantly ≥2% sputum eosinophils in combination with predominantly ≥50% neutrophils showed greater loss of lung function, whereas those with highly variable sputum eosinophils had greater healthcare use.

Clinical relevance of sputum bronchial epithelial cells: A retrospective cross-sectional study

RATIONALE: Bronchial epithelium provides a physical barrier, facilitates mucociliary transport and is involved in regulating various immune responses. Bronchial epithelial cells (BECs) are quantified as part of sputum differential cell counts; however, significance of elevated BEC numbers is not known. We hypothesized that they may be higher in patients whose respiratory symptom is predominantly a cough. OBJECTIVES: Our aim of the study was to characterize patients with increased sputum BECs based on clinical and inflammatory variables. METHODS AND MEASUREMENTS: We conducted a retrospective cross-sectional study utilizing a database containing clinical and inflammatory characteristics of patients who underwent sputum differential cell counts at a tertiary care center in Hamilton, Ontario, Canada. Total and differential cell counts were performed on non-saliva contaminated, DTT-dispersed, sputum cytospins. The BEC high group includes patients with sputum BEC% ≥ 4.4% (90th percentile of normal), whereas the control group includes patients with sputum BEC% < 4.4%. MAIN RESULTS: A total of 7% (239/3420) of patient samples had increased epithelial cells in sputum. Compared to an equal number of control subjects, the patients with elevated BECs were younger, predominately had cough and wheezing, required higher dose of corticosteroids, had higher FEV1 and FVC, and had either eosinophilic or paucigranulocytic inflammatory endotype. There was no significant difference in terms of airway hyperresponsiveness. CONCLUSIONS: Elevated BECs are associated with younger age, higher corticosteroid dose, symptoms of cough and wheezing, and predominately an eosinophilic or paucigranulocytic inflammatory endotype. Mechanisms linking these need to be studied.

The Effect of Age on T-Regulatory Cell Number and Function in Patients With Asthma.

T-regulatory cells (Tregs) play a key role in suppressing effector cells and maintaining self-tolerance. Studies of younger adults and children suggest that insufficient differentiation and functional defects of Tregs may contribute to the development of asthma; however, data from older patients with asthma are limited. To address the effects of aging on the relationship of Treg frequency and function with clinical outcomes, we collected induced sputum (differential cell count and Treg frequency) and peripheral blood (Treg function and frequency) from aged (> 60 years of age) and younger (20–40 years old) patients with asthma. In younger patients, low Treg suppression was associated with significantly higher mean numbers of emergency department (ED) (1.8 vs. 0.17, P = 0.02) and urgent care visits (2.3 vs. 0.17, P = 0.01) for asthma, and decreased asthma control (mean Asthma Control Test [ACT] score, 17 vs. 21.3, P = 0.01) compared to those with high Treg suppression. In older patients, however, a lower Treg function was not significantly associated with ACT scores (18.2 vs. 13.4, P = 0.10), or the number of ED (P = 0.9) or urgent care visits (P = 0.2). Our data suggest that Tregs have a weak relationship with asthma control and clinical asthma outcomes in older patients and differ from findings in younger patients, where Tregs are more likely to play a protective role.

Bacteria and sputum inflammatory cell counts; a COPD cohort analysis

BackgroundThere is evidence that bacterial colonisation in chronic obstructive pulmonary disease (COPD) is associated with increased neutrophilic airway inflammation. This study tested the hypothesis that different bacterial phyla and species cause different inflammatory profiles in COPD patients.MethodsSputum was analysed by quantitative polymerase chain reaction (qPCR) to quantify bacterial load and 16S rRNA gene sequencing to identify taxonomic composition. Sputum differential cell counts (DCC) and blood DCC were obtained at baseline and 6 months. Patients were categorised into five groups based on bacterial load defined by genome copies/ml of ≥ 1 × 104, no colonisation and colonisation by Haemophilus influenzae (H. influenzae), Moraxella catarrhalis (M. catarrhalis), Streptococcus pneumoniae (S. pneumoniae), or > 1 potentially pathogenic microorganism (PPM).ResultsWe observed an increase in sputum neutrophil (%), blood neutrophil (%) and neutrophil–lymphocyte ratio (NLR) in patients colonised with H. influenzae (82.6, 67.1, and 3.29 respectively) compared to those without PPM colonisation at baseline (69.5, 63.51 and 2.56 respectively) (p < 0.05 for all analyses), with similar findings at 6 months. The bacterial load of H. influenzae and Haemophilus determined by qPCR and 16s rRNA gene sequencing respectively, and sputum neutrophil % were positively correlated between baseline and 6 months visits (p < 0.0001, 0.0150 and 0.0002 with r = 0.53, 0.33 and 0.44 respectively).ConclusionsThese results demonstrate a subgroup of COPD patients with persistent H. influenzae colonisation that is associated with increased airway and systemic neutrophilic airway inflammation, and less eosinophilic airway inflammation.

Exploration of the sputum methylome and omics deconvolution by quadratic programming in molecular profiling of asthma and COPD: the road to sputum omics 2.0

BackgroundTo date, most studies involving high-throughput analyses of sputum in asthma and COPD have focused on identifying transcriptomic signatures of disease. No whole-genome methylation analysis of sputum cells has been performed yet. In this context, the highly variable cellular composition of sputum has potential to confound the molecular analyses.MethodsWhole-genome transcription (Agilent Human 4 × 44 k array) and methylation (Illumina 450 k BeadChip) analyses were performed on sputum samples of 9 asthmatics, 10 healthy and 10 COPD subjects. RNA integrity was checked by capillary electrophoresis and used to correct in silico for bias conferred by RNA degradation during biobank sample storage. Estimates of cell type-specific molecular profiles were derived via regression by quadratic programming based on sputum differential cell counts. All analyses were conducted using the open-source R/Bioconductor software framework.ResultsA linear regression step was found to perform well in removing RNA degradation-related bias among the main principal components of the gene expression data, increasing the number of genes detectable as differentially expressed in asthma and COPD sputa (compared to controls). We observed a strong influence of the cellular composition on the results of mixed-cell sputum analyses. Exemplarily, upregulated genes derived from mixed-cell data in asthma were dominated by genes predominantly expressed in eosinophils after deconvolution. The deconvolution, however, allowed to perform differential expression and methylation analyses on the level of individual cell types and, though we only analyzed a limited number of biological replicates, was found to provide good estimates compared to previously published data about gene expression in lung eosinophils in asthma. Analysis of the sputum methylome indicated presence of differential methylation in genomic regions of interest, e.g. mapping to a number of human leukocyte antigen (HLA) genes related to both major histocompatibility complex (MHC) class I and II molecules in asthma and COPD macrophages. Furthermore, we found the SMAD3 (SMAD family member 3) gene, among others, to lie within differentially methylated regions which has been previously reported in the context of asthma.ConclusionsIn this methodology-oriented study, we show that methylation profiling can be easily integrated into sputum analysis workflows and exhibits a strong potential to contribute to the profiling and understanding of pulmonary inflammation. Wherever RNA degradation is of concern, in silico correction can be effective in improving both sensitivity and specificity of downstream analyses. We suggest that deconvolution methods should be integrated in sputum omics analysis workflows whenever possible in order to facilitate the unbiased discovery and interpretation of molecular patterns of inflammation.

Prostaglandin E2 decrease in induced sputum of hypersensitive asthmatics during oral challenge with aspirin.

A special regulatory role for prostaglandin E2 (PGE2 ) has been postulated in nonsteroidal anti-inflammatory drug (NSAID)-exacerbated respiratory disease (NERD). To investigate the effect of systemic aspirin (acetylsalicylic acid) administration on airway PGE2 biosynthesis in induced sputum supernatant (ISS) among subjects with NERD or aspirin-tolerant asthma with chronic rhinosinusitis with nasal polyposis (ATA-CRSwNP), as well as healthy controls (HC). Induced sputum (IS) was collected from patients with NERD (n=26), ATA-CRSwNP (n=17), and HC (n=21) at baseline and after aspirin challenge. Sputum differential cell count and IS supernatant (ISS) levels of prostanoids, PGE2 , 8-iso-PGE2 , tetranor-PGE-M, 8-iso-PGF2 α, and leukotriene C4 , D4 , and E4 , were determined using mass spectrometry. Urinary excretion of LTE4 was measured by ELISA. NERD subjects had elevated sputum eosinophilic count as compared to ATA-CRSwNP and HC (median NERD 9.1%, ATA-CRSwNP 2.1%, and HC 0.4%; P<0.01). Baseline ISS levels of PGE2 were higher in asthmatics as compared to HC at baseline (NERD vs HC P=0.04, ATA-CRSwNP vs HC P<0.05). Post-challenge ISS levels of PGE2 compared to baseline significantly decreased in NERD and HC (P<0.01 and P=0.01), but not in ATA-CRSwNP. In NERD, a similar decrease in PGE2 as in HC resulted from 2.8 times lower dose of aspirin. Aspirin-precipitated bronchoconstriction is associated with a decrease in airway PGE2 biosynthesis. These results support the mechanism of PGE2 biosynthesis inhibition as a trigger for bronchoconstriction in NERD.

Sputum Characteristics among "Chillum” Smoking ‘Sadhus' of Kumbh Mela

A chillum consist a mixture of Cannabis and tobacco.Cannabis, also known as marijuana; it is a psychoactive drug from the Cannabis plant intended for medical or recreational use; nowadays widely being abused for getting high and temporary relaxation. Sputum examination of chillum smokers may indicate harmful effects of chillum smoking on lungs. Objectives of present study were to study the sputum characteristics and differential cell count of chillum smoking Sadhus of Kumbh Mela. Present descriptive observational study was conducted among fifty Sadhus of Kumbh Mela of Nashik. Participants were selected using purposive sampling method. In present study all sputum samples showed raised differential cell count and statistically significant association was observed between differential cell like Neutrophils, Lymphocytes and Macrophages with different content present in chillum.

Sputum Characteristics among �Chillum� Smoking �Sadhus� of Kumbh Mela

A chillum consist a mixture of Cannabis and tobacco.Cannabis, also known as marijuana; it is a psychoactive drug from the Cannabis plant intended for medical or recreational use; nowadays widely being abused for getting high and temporary relaxation. Sputum examination of chillum smokers may indicate harmful effects of chillum smoking on lungs. Objectives of present study were to study the sputum characteristics and differential cell count of chillum smoking Sadhus of Kumbh Mela. Present descriptive observational study was conducted among fifty Sadhus of Kumbh Mela of Nashik. Participants were selected using purposive sampling method. In present study all sputum samples showed raised differential cell count and statistically significant association was observed between differential cell like Neutrophils, Lymphocytes and Macrophages with different content present in chillum.

Saturated fatty acids, obesity, and the nucleotide oligomerization domain–like receptor protein 3 (NLRP3) inflammasome in asthmatic patients

Both obesity and high dietary fat intake activate the nucleotide oligomerization domain-like receptor protein 3 (NLRP3) inflammasome. We aimed to examine NLRP3 inflammasome activity in the airways of obese asthmatic patients after macronutrient overload and in immune cells challenged by inflammasome triggers. Study 1 was a cross-sectional observational study of nonobese (n=51) and obese (n=76) asthmatic adults. Study 2 was a randomized, crossover, acute feeding study in 23 asthmatic adults (n=12 nonobese and n=11 obese subjects). Subjects consumed 3 isocaloric meals on 3 separate occasions (ie, saturated fatty acid, n-6 polyunsaturated fatty acid, and carbohydrate) and were assessed at 0 and 4hours. For Studies 1 and 2, airway inflammation was measured based on sputum differential cell counts, IL-1β protein levels (ELISA), and sputum cell gene expression (Nanostring nCounter). In Study 3 peripheral blood neutrophils and monocytes were isolated by using Ficoll density gradient and magnetic bead separation and incubated with or without palmitic acid, LPS, or TNF-α for 24hours, and IL-1β release was measured (ELISA). In Study 1 NLRP3 and nucleotide oligomerization domain 1 (NOD1) gene expression was upregulated, and sputum IL-1β protein levels were greater in obese versus nonobese asthmatic patients. In Study 2 the saturated fatty acid meal led to increases in sputum neutrophil percentages and sputum cell gene expression of Toll-like receptor 4 (TLR4) and NLRP3 at 4hours in nonobese asthmatic patients. In Study 3 neutrophils and monocytes released IL-1β when challenged with a combination of palmitic acid and LPS or TNF-α. The NLRP3 inflammasome is a potential therapeutic target in asthmatic patients. Behavioral interventions that reduce fatty acid exposure, such as weight loss and dietary saturated fat restriction, warrant further exploration.

Menopausal asthma–much ado about nothing? An observational study

ABSTRACTObjective: Menopausal asthma is considered a distinct asthma phenotype. Our aim was to identify potential specific features of asthma in postmenopausal women in a cohort of Polish females. Methods: Asthma severity and control, pulmonary function, exhaled nitric oxide (FENO), peripheral blood and induced sputum (IS) differential cell count were compared in three groups: women with premenopausal asthma (group 1), menopausal women with pre-existing asthma (group 2A) and menopausal women with asthma onset in the perimenopausal or menopausal period (group 2B). Results: We enrolled 27 women to group 1, 13 to group 2A and 16 to group 2B. Asthma severity and control, blood eosinophil count and FENO did not differ among the groups. Menopausal women had a higher incidence of irreversible airway obstruction (84.6% in group 2A and 56.2% in group 2B vs. 22.2% in group 1, p < 0.001 and p = 0.03, respectively). The proportion of patients with sputum eosinophilia was highest in menopausal women with pre-existing asthma, although the difference did not reach statistical significance (88.9% in group 2A vs. 66.7% in group 2B and 65.0% in group 1, respectively, p = 0.86). Conclusions: Menopausal women with asthma are characterized by an increased incidence of irreversible airway obstruction regardless of disease duration. This may indicate that age may contribute to pulmonary function impairment in asthmatic women independently of their hormonal status at the time of asthma diagnosis. Our results failed to confirm the presence of specific asthma features which would allow to distinguish the phenotype of menopausal asthma.

Does chronic physical activity level modify the airway inflammatory response to an acute bout of exercise in the postprandial period?

Recent studies have confirmed that a single high-fat meal (HFM) leads to increased airway inflammation. However, exercise is a natural anti-inflammatory and may modify postprandial airway inflammation. The postprandial airway inflammatory response is likely to be modified by chronic physical activity (PA) level. This study investigated whether chronic PA modifies the airway inflammatory response to an acute bout of exercise in the postprandial period in both insufficiently active and active subjects. Thirty-nine nonasthmatic subjects (20 active, 13 males/7 females) who exceeded PA guidelines (≥150 min moderate-vigorous PA/week) and 19 insufficiently active (6 males/13 females) underwent an incremental treadmill test to exhaustion to determine peak oxygen uptake. Subjects were then randomized to a condition (COND), either remaining sedentary (CON) or exercising (EX) post-HFM. Exercise was performed at the heart rate corresponding to 60% peak oxygen uptake on a treadmill for 1 h post-HFM (63% fat, 10 kcal/kg body weight). Blood lipids and exhaled nitric oxide (eNO: marker of airway inflammation) were measured at baseline and 2 h and 4 h post-HFM. Sputum differential cell counts were performed at baseline and 4 h post-HFM. The mean eNO response for all groups increased at 2 h post-HFM (∼6%) and returned to baseline by 4 h (p = 0.03). There was a time × COND interaction (p = 0.04), where EX had a greater eNO response at 4 h compared with CON. Sputum neutrophils increased at 4 h post-HFM (p < 0.05). These findings suggest that airway inflammation occurs after an HFM when exercise is performed in the postprandial period, regardless of habitual activity level.

The duration of cough in patients with H1N1 influenza.

BackgroundCough is one of common symptoms of influenza, the cough duration and prevalence of postinfectious cough (PIC) after viral upper respiratory tract infection has not been well described.ObjectivesWe aim to investigate the duration of cough and prevalence of PIC and its relation with acute symptoms, airway inflammation and cough sensitivity in patients with H1N1 influenza.MethodsPatients with acute symptoms of H1N1 influenza were enrolled and followed up until cough relived. Spirometry, induced sputum test, capsaicin challenge test were conducted in patients with PIC. Cough sensitivity was presented as logarithm of provocative concentration inducing five or more coughs (logC5).ResultsA total of 141 cases with H1N1 influenza were enrolled. In patients with H1N1 influenza, 97.2% of them complained cough. The duration of cough was as following: <1 week (73.0%); 1–2 weeks (7.8%); 2–3 weeks (7.8%); ≥3 weeks (8.5%). Twelve (8.5%) patients had cough lasting more than 3 weeks (PIC), 4 (2.8%) patients developed chronic cough (>8 weeks). Acute symptoms, spirometry, bronchial responsiveness and sputum differential cell count were similar between patients with PIC and those without PIC, however, there was a higher prevalence of previous PIC (58.3% vs 14.7%, P < 0.05) and elevated cough sensitivity (lgC5: 1.18 ± 0.58 vs 2.73 ± 0.33, P < 0.01) in patients with PIC as compared with the patients without PIC.ConclusionsAcute cough is common in patients with H1N1 PIC, only a few of patients develop chronic cough. Acute symptoms cannot predict PIC which is related with previous PIC and increased cough sensitivity.

An Asthmatic Population With High Macrophages in Their Sputum Differential Cell Counts

An Asthmatic Population With High Macrophages in Their Sputum Differential Cell Counts

The Effect of Physical Activity on Post-Prandial Triglycerides and Airway Inflammation Following a High-Fat Meal

Recent studies have confirmed that a single high-fat meal (HFM) leads to increased triglycerides and airway inflammation, which may be attenuated by chronic exercise training or an acute bout of exercise. PURPOSE: To investigate whether an acute bout of moderate intensity exercise following a HFM would attenuate post-prandial lipemia (PPL) and airway inflammation in both inactive and active subjects. METHODS: Thirty-five, college-aged subjects took part in the study. Twenty subjects (13M/7F) exceeded physical activity (PA) guidelines (≥150 minutes moderate-vigorous PA/week) and fifteen subjects (5M/10F) did not meet PA guidelines. At the first visit, subjects underwent DEXA scans, pulmonary function tests, and an incremental test to exhaustion to determine VO2peak. On the second visit, subjects were randomized to the control condition ((ACT CON (n=10) or IN CON (n=6)) or the exercise condition ((ACT EX (n=10) or IN EX (n=9)). The EX condition performed the exercise bout (60%VO2peak; ∼30-135 minutes) on a treadmill 45 minutes post-HFM (63% fat, 10kcal/kg of body weight). Blood lipids and exhaled nitric oxide (eNO) were measured at baseline and 2 h and 4 h post-HFM. Sputum differential cell counts were performed at baseline and 4 h post-HFM to assess airway inflammation. RESULTS: Triglycerides increased significantly in ACT CON and ACT EX 2 h (∼54%, p=0.001; ∼66%, p=0.013, respectively) and 4 h (∼107%, p=0.001; ∼119%, p=0.004, respectively) post-HFM and also IN CON at 4 h (∼44%; p= 0.004) and IN EX (2 h, ∼62%, p= 0.001; 4 h, ∼93%, p=0.001) post-HFM. Area under the curve (AUC) was significantly lower in ACT EX (p<0.001) than ACT CON and IN EX, but was not significantly different in IN CON versus IN EX (p>0.05). Exhaled nitric oxide increased non-linearly from baseline to 2 and 4 h post-HFM in ACT (p<0.05, η2=0.36) and IN (<0.05), but was not dependent on condition. CONCLUSION: Preliminary findings suggest that in both active and inactive subjects, an exercise bout after a HFM did not reduce post-prandial airway inflammation. However, the exercise bout was effective in reducing PPL in active subjects, but not inactive subjects. Post-prandial airway inflammation occurred following a high-fat meal regardless of a reduction in post-prandial triglycerides.

P49 Can The Asthma Control Questionnaire (acq) And/or The Blood Eosinophil Count Accurately Detect Sputum Eosinophilia?

Induced sputum differential cell counts provide important information about airway inflammation and future risk in asthma, but are not universally available. We set out to ascertain the sensitivity and specificity of the Juniper Asthma Control Questionnaire (ACQ) and/or the peripheral blood eosinophil count to detect a sputum eosinophil count >3%. Methods We performed a retrospective, cross-sectional study of 165 subjects with asthma, aged 18–80, prescribed as-required bronchodilators, long-acting β-agonists, 0–4000 mcg inhaled beclometasone diproprionate equivalent or maintenance oral steroids (0–20 mg prednisolone) from our database. Current smokers were excluded. Spirometry, FeNO at 50 ml flow, sputum induction, Asthma Control Questionnaire (ACQ) and blood eosinophils (BEos) were recorded. Induced sputum eosinophils (SEos) defined eosinophilic (EA, SEos ≥3%) and non-eosinophilic (NEA, SEos 10 transformed and groups were compared with t-tests or Mann-Whitney-U using STATA and GraphPad. Receiver operating characteristic (ROC) curves determined cut-points of ACQ and BEos that identified SEos ≥3%, and these were tested retrospectively in a second population of adults with asthma (n = 48, 40% EA, mean FEV 1 82% predicted). Results The 31% with EA had a lower FEV 1 % predicted and FEV 1 /FVC ratio, a higher FeNO, BEos and ACQ compared to NEA (Table 1). There was a significant correlation between ACQ and sputum eosinophils (Pearson r 0.32, p The optimal cut-points of ACQ and BEos identifying a SEos of 3% were an ACQ >1.57 (sensitivity 55%, specificity 78%) and BEos ≥0.22 × 10 9 /L (sensitivity 84% and specificity 68%) respectively. Individuals with both ACQ of ≥1.57 AND a BEos of ≥0.22 the sensitivity was 45% and specificity of 100% for a SEos of 3%. Testing in the second population the ACQ≥1.57 had a sensitivity 47% and specificity 86% for SEos >3% while BEos ≥0.22 had sensitivity 68% and specificity 45%. The combination of ACQ ≥1.57 AND BEos ≥0.22 had a lower sensitivity of 37% but a high specificity of 93% and could be useful to exclude rather than identify the presence of sputum eosinophilia. Conclusion Combined testing using ACQ and BEos only identifies eosinophilia in 37% and can’t replace induced sputum, but may be useful to exclude sputum eosinophilia and prospective study is warranted.

Induced sputum eicosanoids during aspirin bronchial challenge of asthmatic patients with aspirin hypersensitivity.

Altered metabolism of eicosanoids is a characteristic finding in aspirin-exacerbated respiratory disease (AERD). Bronchial challenge with lysyl-aspirin can be used as a confirmatory diagnostic test for this clinical condition. Induced sputum allows to measure mediators of asthmatic inflammation in bronchial secretions. To investigate the influence of inhaled lysyl-aspirin on sputum supernatant concentration of eicosanoids during the bronchial challenge test. Subjects with asthma hypersensitive to nonsteroidal anti-inflammatory drugs were compared with aspirin-tolerant asthmatic controls. Induced sputum was collected before and following bronchial challenge with lysyl-aspirin. Sputum differential cell count and sputum supernatant concentrations of selected lipoxygenases products: 5-,12-,15-hydroxyeicosatetraenoic acid, cysteinyl leukotrienes, leukotriene B4 , 11-dehydro-thromboxane B2 , and prostaglandins E2 , D2 , and F2α and their metabolites, were measured using validated methods of chromatography-mass spectrometry. Aspirin precipitated bronchoconstriction in all AERD subjects, but in none of the aspirin-tolerant asthmatics. Phenotypes of asthma based on the sputum cytology did not differ between the groups. Baseline sputum eosinophilia correlated with a higher leukotriene D4 (LTD4 ) and leukotriene E4 (LTE4 ) concentrations. LTC4 , PGE2 , and 11-dehydro-TXB2 did not differ between the groups, but levels of LTD4 , LTE4 , and PGD2 were significantly higher in AERD group. Following the challenge, LTD4 and LTE4 increased, while PGE2 and LTB4 decreased in AERD subjects only. During the bronchial challenge, decrease in PGE2 and its metabolite is accompanied by a surge in bronchoconstrictory cysteinyl leukotrienes produced at the expense of LTB4 in AERD subjects. Bronchial PGE2 inhibition in AERD seems specific and sensitive to a low dose of aspirin.