Non-severe Asthma Research Articles

Clinical utility of neutrophil-to-lymphocyte ratio in the assessment of asthma and its severity

Homoeostasis of the immune system is regulated by cell-mediated immune-inflammatory responses. The shift of the immune response is clearly reflected by the cellular ratios than individual cell count. Neutrophil-to-lymphocyte ratio (NLR) serves as a good cell-based marker of inflammation, which is being investigated in various inflammation-related diseases. Asthma which is a chronic inflammatory airway disease associated with hyperresponsiveness, recurrent attacks, wheezing, coughing and chest tightness coupled with multicellular influx of granulocytes with mild-to-severe symptoms of and altered NLR. The aim of the present study was to explore the existing literature on NLR in asthma to realise its clinical utility in asthma and its severity. We searched online database resources 'PubMed' and 'Google Scholar' up till now (20th May, 2022), and studies were included as per eligibility criteria. The data collected were reviewed and analysed. The study included eight published articles on NLR and asthma. The analysis revealed that there is a significant difference in mean NLR between healthy controls and asthma patients (1.57 ± 0.35 vs. 2.36 ± 0.30; P < 0.0001); further, severe versus non-severe asthma cases showed the highest mean NLR as compared to non-severe cases (4.82 ± 2.85 vs. 2.13 ± 0.17; P < 0.0001). Receiver operating curve analysis of NLR showed excellent discriminating capacity, for health versus disease and controlled versus uncontrolled. This mini review suggests NLR can serve as a non-expensive, non-invasive monitoring tool to reduce the risk of exacerbation and hospitalisation. However, large studies are warranted.

Airway microbiota and immune mediator relationships differ in obesity and asthma

Asthma and obesity are both complex conditions characterized by chronic inflammation, and obesity-related severe asthma has been associated with differences in the microbiome. However, whether the airway microbiome and microbiota-immune response relationships differ between obese persons with or without nonsevere asthma is unestablished. We compared the airway microbiome and microbiota-immune mediator relationships between obese and nonobese subjects, with and without mild-moderate asthma. We performed cross-sectional analyses of the airway (induced sputum) microbiome and cytokine profiles from blood and sputum using 16S ribosomal RNA gene and internal transcribed spacer region sequencing to profile bacteria and fungi, and multiplex immunoassays. Analysis tools included QIIME 2, linear discriminant analysis effect size (aka LEfSe), Piphillin, and Sparse inverse covariance estimation for ecological association inference (aka SPIEC-EASI). Obesity, irrespective of asthma status, was associated with significant differences in sputum bacterial community structure and composition (unweighted UniFrac permutational analysis of variance, P= .02), including a higher relative abundance of Prevotella, Gemella, and Streptococcus species. Among subjects with asthma, additional differences in sputum bacterial composition and fungal richness were identified between obese and nonobese individuals. Correlation network analyses demonstrated differences between obese and nonobese asthma in relationships between cytokine mediators, and these together with specific airway bacteria involving blood PAI-1, sputum IL-1β, GM-CSF, IL-8, TNF-α, and several Prevotella species. Obesity itself is associated with an altered sputum microbiome, which further differs in those with mild-moderate asthma. The distinct differences in airway microbiota and immune marker relationships in obese asthma suggest potential involvement of airway microbes that may affect mechanisms or outcomes of obese asthma.

Epidemiology of patients with asthma in Korea: Analysis of the NHISS database 2006–2015

BackgroundThere has been a concerning increase in the prevalence and socioeconomic burden of asthma in Korea. Korea's National Health Insurance System (NHIS) covers insurance payment and claims management for all Koreans. Using National Health Insurance Sharing Service (NHISS) claims data. This study aimed to investigate patterns of healthcare utilization and direct cost in patients with asthma over a 10-year period. MethodsIn this retrospective population-based study, we examined NHISS claims records between July 2005 and June 2016 and investigated healthcare utilization among patients with asthma based on age group and severity of disease (non-severe asthma [NSA] and severe asthma [SA]). ResultsFrom 2006 to 2015, the total number of patients with asthma in Korea steadily increased from 743 968 to 2 286 309, with a corresponding increase in prevalence from 1.62% to 4.74%. The proportion of patients with SA decreased from 3.16% in 2006 to 1.56% in 2015; the proportion was consistently higher in men than in women. In addition, patients with SA had a higher cost per outpatient visit than patients with NSA, and the number of outpatient visits per year increased. The inhaled corticosteroid (ICS) prescription rate among patients with asthma decreased from 22.9% in 2006 to 15.7% in 2015. Furthermore, for a period of 10 years, more than 40% of patients with SA have been prescribed short-acting β-2 agonists (SABAs). ConclusionsAlthough patients with SA comprised a small proportion of patients with asthma, they incurred greater medical costs per person. The pharmaceutical prescription pattern indicated a lack of ICS-based prescriptions and frequent SABA prescriptions.

Genetic analyses of chr11p15.5 region identify MUC5AC-MUC5B associated with asthma-related phenotypes

Objective Genome-wide association studies (GWASs) have identified single nucleotide polymorphisms (SNPs) in chr11p15.5 region associated with asthma and idiopathic interstitial pneumonias (IIPs). We sought to identify functional genes for asthma by combining SNPs and mRNA expression in bronchial epithelial cells (BEC) in the Severe Asthma Research Program (SARP). Methods Correlation analyses of mRNA expression of six candidate genes (AP2A2, MUC6, MUC2, MUC5AC, MUC5B, and TOLLIP) and asthma phenotypes were performed in the longitudinal cohort (n = 156) with RNAseq in BEC, and replicated in the cross-sectional cohort (n = 155). eQTL (n = 114) and genetic association analysis of asthma severity (426 severe vs. 531 non-severe asthma) were performed, and compared with previously published GWASs of IIPs and asthma. Results Higher expression of AP2A2 and MUC5AC and lower expression of MUC5B in BEC were correlated with asthma, asthma exacerbations, and T2 biomarkers (P < 0.01). SNPs associated with asthma and IIPs in previous GWASs were eQTL SNPs for MUC5AC, MUC5B, or TOLLIP, however, they were not in strong linkage disequilibrium. The risk alleles for asthma or protective alleles for IIPs were associated with higher expression of MUC5AC and lower expression of MUC5B. rs11603634, rs12788104, and rs28415845 associated with moderate-to-severe asthma or adult onset asthma in previous GWASs were not associated with asthma severity (P > 0.8). Conclusions SNPs associated with asthma in chr11p15.5 region are not associated with asthma severity neither with IIPs. Higher expression of MUC5AC and lower expression of MUC5B are risk for asthma but protective for IIPs.

Blood Inflammatory-like and Lung Resident-like Eosinophils Affect Migration of Airway Smooth Muscle Cells and Their ECM-Related Proliferation in Asthma

Airway remodeling is a hallmark feature of asthma, and one of its key structural changes is increased airway smooth muscle (ASM) mass and disturbed extracellular matrix (ECM) homeostasis. Eosinophil functions in asthma are broadly defined; however, we lack knowledge about eosinophil subtypes' interaction with lung structural cells and their effect on the airway's local microenvironment. Therefore, we investigated the effect of blood inflammatory-like eosinophils (iEOS-like) and lung resident-like eosinophils (rEOS-like) on ASM cells via impact on their migration and ECM-related proliferation in asthma. A total of 17 non-severe steroid-free allergic asthma (AA), 15 severe eosinophilic asthma (SEA) patients, and 12 healthy control subjects (HS) were involved in this study. Peripheral blood eosinophils were enriched using Ficoll gradient centrifugation and magnetic separation, subtyped by using magnetic separation against CD62L. ASM cell proliferation was assessed by AlamarBlue assay, migration by wound healing assay, and gene expression by qRT-PCR analysis. We found that blood iEOS-like and rEOS-like cells from AA and SEA patients' upregulated genes expression of contractile apparatus proteins, COL1A1, FN, TGF-β1 in ASM cells (p < 0.05), and SEA eosinophil subtypes demonstrated the highest effect on sm-MHC, SM22, and COL1A1 gene expression. Moreover, AA and SEA patients' blood eosinophil subtypes promoted migration of ASM cells and their ECM-related proliferation, compared with HS (p < 0.05) with the higher effect of rEOS-like cells. In conclusion, blood eosinophil subtypes may contribute to airway remodeling by upregulating contractile apparatus and ECM component production in ASM cells, further promoting their migration and ECM-related proliferation, with a stronger effect of rEOS-like cells and in SEA.

Организация помощи пациентам с тяжелой бронхиальной астмой, нуждающимся в лечении генно-инженерными биологическими препаратами, в Санкт-Петербурге

The proportion of patients with severe asthma among asthma patients is approximately 5–10%. Severe asthma is associated with a heavy economic burden and remains a major public health problem. Patients with severe asthma require significant use of healthcare resources, i.e., the rate of exacerbation-related hospital admissions and unscheduled emergency department visits is two- and three-fold, respectively, higher than that in non-severe asthma. The discovery of the molecular mechanisms of asthma pathogenesis allowed the development of biological genetically engineering therapies that provide an effective therapeutic option for severe asthma by improving the quality of life by reducing the rate of exacerbations, improving lung function, and reducing the need for systemic steroids with a good safety profile. Targets for existing biological agents include IgE, interleukins (IL)-4, 5, and 13, and thymic stromal lymphopoietin. These drugs include omalizumab, mepolizumab, reslizumab, benralizumab, dupilumab, and tezepelumab, which were recently registered in Russia. Pharmacoeconomic analysis demonstrates a reduction in the costs of managing patients with severe asthma who receive targeted therapy despite the financial costs of its implementation. The choice of targeted drug is always a complex task that requires careful assessment of clinical, anamnestic, and laboratory parameters. In addition, monoclonal antibodies are an expensive treatment option. Currently, centers for genetically engineering biological therapies are being created to achieve effective routing and continuity in the management of these patients and to minimize financial costs. KEYWORDS: severe asthma, eosinophilic phenotype, genetically engineered biological therapy (GEBT), registry, medical commission, targeted therapy, monoclonal antibodies, type 2 immune response. FOR CITATION: Titova O.N., Volchkov V.A., Kuzubova N.A., Sklyarova D.B. Organization of medical care for patients with severe asthma requiring genetically engineered biological therapies in St. Petersburg. Russian Medical Inquiry. 2023;7(8):493–497 (in Russ.). DOI: 10.32364/2587-6821-2023-7-8-3.

IL-5 and GM-CSF, but Not IL-3, Promote the Proliferative Properties of Inflammatory-like and Lung Resident-like Eosinophils in the Blood of Asthma Patients.

Blood eosinophils can be described as inflammatory-like (iEOS-like) and lung-resident-like (rEOS-like) eosinophils. This study is based on the hypothesis that eosinophilopoetins such as interleukin (IL)-3 and IL-5 and granulocyte-macrophage colony-stimulating factor (GM-CSF) alter the proliferative properties of eosinophil subtypes and may be associated with the expression of their receptors on eosinophils. We investigated 8 individuals with severe nonallergic eosinophilic asthma (SNEA), 17 nonsevere allergic asthma (AA), and 11 healthy subjects (HS). For AA patients, a bronchial allergen challenge with Dermatophagoides pteronyssinus was performed. Eosinophils were isolated from peripheral blood using high-density centrifugation and magnetic separation methods. The subtyping of eosinophils was based on magnetic bead-conjugated antibodies against L-selectin. Preactivation by eosinophilopoetins was performed by incubating eosinophil subtypes with IL-3, IL-5, and GM-CSF, and individual combined cell cultures were prepared with airway smooth muscle (ASM) cells. ASM cell proliferation was assessed using an Alamar blue assay. The gene expression of eosinophilopoetin receptors was analyzed with a qPCR. IL-5 and GM-CSF significantly enhanced the proliferative properties of iEOS-like and rEOS-like cells on ASM cells in both SNEA and AA groups compared with eosinophils not activated by cytokines (p < 0.05). Moreover, rEOS-like cells demonstrated a higher gene expression of the IL-3 and IL-5 receptors compared with iEOS-like cells in the SNEA and AA groups (p < 0.05). In conclusion: IL-5 and GM-CSF promote the proliferative properties of iEOS-like and rEOS-like eosinophils; however, the effect of only IL-5 may be related to the expression of its receptors in asthma patients.

Apurinic/apyrimidinic endonuclease 1 mRNA level in peripheral blood neutrophils is associated with asthma

Apurinic/apyrimidinic endonuclease 1 (APE1) is a multifunc-tional key protein. Recent studies suggest APE1 is closely associated with in-flammatory response, but its role in asthma remains unknown. We recruited 116 patients with asthma, including 50 with severe asthma (NSA) and 66 with non-severe asthma (SA), and 140 controls. Serum APE1 was detected using the ELISA method. APE1 mRNA in peripheral blood neutrophils and eosinophils were detected using real-time PCR assays. Compared to healthy controls, we observed significant elevations of serum APE1 mRNA levels in peripheral neu-trophils (~1.75 folds increase, p&lt;0.05) and eosinophils (~2.2 folds increase, p&lt;0.05) in patients with asthma. The peripheral blood neutrophil APE1 mRNA can distinguish asthmatic patients from healthy controls with the area under the curve (AUC) 0.893 and a 95% confidence interval (CI) 0.847-0.938 (p &lt; 0.001). Also the APE1 mRNA can identify severe asthma from non-severe asth-ma (AUC 0.759, 95% CI, 0.674-0.846; p &lt; 0.001). However, The serum APE1 and eosinophil mRNA levels did not correlate with asthma incidence and sever-ity. Our finding confirms the association between APE1 and asthma and sug-gests that peripheral blood neutrophil APE1 mRNA may be used as a marker for this condition.

Awareness of asthma and its management in primary school teachers in Baghdad, Iraq.

Background: Asthma is a major global health issue characterized by chronic airway inflammation. It is linked to a high disease burden and disproportionately high healthcare utilization in severe, uncontrolled cases compared to non-severe asthma. We aimed to conduct this survey among primary school teachers in Baghdad, Iraq, to assess their level of knowledge about asthma and confidence in managing school children with asthma. Methods: This was a cross-sectional study conducted in Baghdad. The study adopted a questionnaire for assessing the asthma knowledge and confidence scores regarding the management of asthmatic children. This questionnaire contained 29-multiple true-false questions based on different aspects, including facts about asthma and the management of asthma by teachers. The questionnaire for testing teachers' knowledge of asthma and confidence scores regarding the management of asthmatic children was distributed to 150 teachers. Results: The questionnaire was completed by 103 teachers.. Approximately 71% of teachers answered the question 'What are the three main symptoms of asthma?' correctly i.e., answering either one symptom (35.9%) or two symptoms (35.9%) correctly. A relatively smaller number of teachers (16.5%) mentioned all three symptoms correctly. Across the 29-multiple true-false questions, more than 75% of teachers answered 11 questions correctly, 50-74% of teachers answered the rest of eight questions appropriately, and <50% of teachers answered the remaining 10 questions properly. The statistical evaluation indicated that the mean total knowledge score about asthma for all the teachers was 20.27 ± 2.97 and the mean total confidence score regarding the management of asthmatic children was 72.44 ± 13.61. Conclusions: This study suggests that teachers from the schools in Baghdad appear to be self-confident in their ability and knowledge to help and manage children with asthma.

Urinary total conjugated 3-bromotyrosine, asthma severity, and exacerbation risk.

Asthma is an inflammatory disease of the airways characterized by eosinophil recruitment, eosinophil peroxidase release, and protein oxidation through bromination, which following tissue remodeling results in excretion of 3-bromotyrosine. Predicting exacerbations and reducing their frequency is critical for the treatment of severe asthma. In this study, we aimed to investigate whether urinary total conjugated bromotyrosine can discriminate asthma severity and predict asthma exacerbations. We collected urine from participants with severe (n = 253) and nonsevere (n = 178) asthma, and the number of adjudicated exacerbations in 1-yr longitudinal follow-up was determined among subjects enrolled in the Severe Asthma Research Program, a large-scale National Institutes of Health (NIH)-funded consortium. Urine glucuronidated bromotyrosine and total conjugated forms were quantified by hydrolysis with either glucuronidase or methanesulfonic acid, respectively, followed by liquid chromatography-tandem mass spectrometry analyses of free 3-bromotyrosine. Blood and sputum eosinophils were also counted. The majority of 3-bromotyrosine in urine was found to exist in conjugated forms, with glucuronidated bromotyrosine representing approximately a third, and free bromotyrosine less than 1% of total conjugated bromotyrosine. Total conjugated bromotyrosine was poorly correlated with blood (r2 = 0.038) or sputum eosinophils (r2 = 0.0069). Compared with participants with nonsevere asthma, participants with severe asthma had significantly higher urinary total conjugated bromotyrosine levels. Urinary total conjugated bromotyrosine was independently associated with asthma severity, correlated with the number of asthma exacerbations, and served as a predictor of asthma exacerbation risk over 1-yr of follow-up.

Expression of Eosinophilic Subtype Markers in Patients with Kawasaki Disease.

Purpose: Eosinophils may rise to a higher level in the acute phase of Kawasaki disease (KD) both before and after intravenous immunoglobulin (IVIG) therapy. A substantial body of research was carried out on the association between KD and allergic diseases. Eosinophils play an important role in type 2 inflammation. Recent studies have shown that there are two distinct subtypes of eosinophils. In addition to their role in inflammation, lung-resident eosinophils (rEOS) also regulate homeostasis. Inflammatory eosinophils (iEOS) reflect type 2 inflammation in tissues. iEOS were considered the primary eosinophils in non-severe allergic asthma, while rEOS were thought to be the primary eosinophils in severe non-allergic eosinophilic asthma. This case–control study aimed to investigate the marker expression of eosinophilic subtypes in KD patients. Materials and Methods: The marker expressions of eosinophilic subtypes in the leukocytes of patients with KD were evaluated by the recently established KDmarkers online tool, a web server including gene expression data. Finally, the results were validated with a quantitative reverse transcriptase polymerase chain reaction (RT-PCR). We analyzed the mRNA expression levels of SELL and IL10RA in leukocytes from KD patients and febrile children. Results: Included in our screening tools were transcriptome arrays, which provided clues showing the importance of rEOS, whose role was identified by three genes (lower IL10RA, higher SELL, and SERPINB1 than controls). In contrast, the iEOS representative gene CD101 was not elevated in KD. It was found that the gene IL10RA, a marker of inflammatory eosinophilic leukocytes, was more highly expressed in the leukocytes of KD patients (n = 43) than febrile controls (n = 32), especially those without coronary artery lesions (CAL) (n = 26). Before treatment, SELL expression was higher in leukocytes of CAL patients (CAL, 1.33 ± 0.18, n = 39; non-CAL, 0.87 ± 0.12, n = 55; p = 0.012). SELL was significantly higher after half a year compared to febrile controls. Conclusions: To our knowledge, this is the first study to demonstrate that KD patients have increased SELL than febrile controls after 6 months of treatment. We present evidence here that dynamically different eosinophilic involvement exists between KD patients with and without CAL. The role of eosinophilic subtypes in KD patients warrants further investigation.

Determinants of lung function across childhood in the Severe Asthma Research Program (SARP) 3

Children with asthma are at risk for low lung function extending into adulthood, but understanding of clinical predictors is incomplete. We sought to determine phenotypic factors associated with FEV1 throughout childhood in the Severe Asthma Research Program 3 pediatric cohort. Lung function was measured at baseline and annually. Multivariate linear mixed-effects models were constructed to assess the effect of baseline and time-varying predictors of prebronchodilator FEV1 at each assessment for up to 6 years. All models were adjusted for age, predicted FEV1 by Global Lung Function Initiative reference equations, race, sex, and height. Secondary outcomes included postbronchodilator FEV1 and prebronchodilator FEV1/forced vital capacity. A total of 862 spirometry assessments were performed for 188 participants. Factors associated with FEV1 include baseline Feno (B, -49 mL/log2 PPB; 95% CI, -92 to -6), response to a characterizing dose of triamcinolone acetonide (B, -8.4 mL/1% change FEV1 posttriamcinolone; 95% CI, -12.3 to -4.5), and maximal bronchodilator reversibility (B, -27 mL/1% change postbronchodilator FEV1; 95% CI, -37 to -16). Annually assessed time-varying factors of age, obesity, and exacerbation frequency predicted FEV1 over time. Notably, there was a significant age and sex interaction. Among girls, there was no exacerbation effect. For boys, however, moderate (1-2) exacerbation frequency in the previous 12 months was associated with -20 mL (95% CI, -39 to -2) FEV1 at each successive year. High exacerbation frequency (≥3) 12 to 24 months before assessment was associated with -34 mL (95% CI, -61 to -7) FEV1 at each successive year. In children with severe and nonsevere asthma, several clinically relevant factors predict FEV1 over time. Boys with recurrent exacerbations are at high risk of lower FEV1 through childhood.

Association of Serum Vitamin D Levels, IL-17a Levels with Disease Severity among 3- to 18-year-old Children with Asthma in the Philippine General Hospital

Background. Previous studies show that Vitamin D has an inverse relationship with asthma severity, symptoms, exacerbations, medication usage, and a direct relationship with lung function. IL-17A was found to be increased in asthmatics, which was inhibited by Vitamin D. Associations found between vitamin D, IL-17A, and asthma may support the future role of vitamin D in the treatment of asthma in children.&#x0D; Objective. To compare vitamin D and IL-17A levels between children with and without asthma and determine their association with asthma severity&#x0D; Study Design. Cross-sectional study&#x0D; Methods. There were 44 participants, aged 3 to 18 years: 22 with asthma (12 non-severe, 10 severe) and 22 without asthma. Participants with any disease-altering vitamin D metabolism, intake of vitamin D supplementation, and recent infection were excluded. Serum vitamin D and IL-17A levels were measured in all participants.&#x0D; Results. There was no significant difference in mean vitamin D levels between participants with asthma (29.6 ± 12.6 ng/mL) and without asthma (27.6 ± 9.5 ng/mL) (p = 0.55) as well as between participants with non-severe asthma (29.8 ± 14.0 ng/mL) and severe asthma (29.4 ± 11.5 ng/mL) (p = 0.95). The overall prevalence of hypovitaminosis D (&lt; 30ng/mL) is 61.4%; 59.1% among those with asthma and 63.6% without asthma. The prevalence of vitamin D insufficiency and/or deficiency did not significantly differ between those with and without asthma (all p-value &gt; 0.05); prevalence ratios were: 1.05 for vitamin D insufficiency, 0.58 for vitamin deficiency, and 0.92 for vitamin D insufficiency and deficiency combined. There was also no significant difference in the prevalence of vitamin D insufficiency and/or deficiency between severe and non-severe asthma (all p-values &gt; 0.05), with prevalence ratios: 0.74 for vitamin D insufficiency, 0.50 for vitamin D deficiency, and 0.75 for vitamin D insufficiency and deficiency combined. Serum IL-17A levels were below the minimum detectable levels in 96% of the participants using the MILLIPLEX Map Human TH17 Magnetic Band Panel; hence, could not be analyzed.&#x0D; Conclusion. The mean serum vitamin D levels do not differ between children with asthma and healthy controls. There was no significant relationship between mean vitamin D levels and asthma severity. There was no association between the prevalence of vitamin D insufficiency and/or deficiency and asthma and its severity. The overall prevalence of hypovitaminosis D in this study is 61.4%. Serum IL-17A levels were undetectable in 96% of the study population.

Awareness of asthma and its management in primary school teachers in Baghdad, Iraq

Background: Asthma is a major global health issue characterized by chronic airway inflammation. It is linked to a high disease burden and disproportionately high healthcare utilization in severe, uncontrolled cases compared to non-severe asthma. We aimed to conduct this survey among primary school teachers in Baghdad, Iraq, to assess their level of knowledge about asthma and confidence in managing school children with asthma. Methods: This was a cross-sectional study conducted in Baghdad. The study adopted a questionnaire for assessing the asthma knowledge and confidence scores regarding the management of asthmatic children. This questionnaire contained 29-multiple true-false questions based on different aspects, including facts about asthma and the management of asthma by teachers. The questionnaire for testing teachers’ knowledge of asthma and confidence scores regarding the management of asthmatic children was distributed to 150 teachers. Results: The questionnaire was completed by 103 teachers.. Approximately 71% of teachers answered the question ‘What are the three main symptoms of asthma?’ correctly i.e., answering either one symptom (35.9%) or two symptoms (35.9%) correctly. A relatively smaller number of teachers (16.5%) mentioned all three symptoms correctly. Across the 29-multiple true-false questions, more than 75% of teachers answered 11 questions correctly, 50-74% of teachers answered the rest of eight questions appropriately, and <50% of teachers answered the remaining 10 questions properly. The statistical evaluation indicated that the mean total knowledge score about asthma for all the teachers was 20.27 ± 2.97 and the mean total confidence score regarding the management of asthmatic children was 72.44 ± 13.61. Conclusions: This study suggests that teachers from the schools in Baghdad appear to be self-confident in their ability and knowledge to help and manage children with asthma.

High proportion of inflammatory CD62Llow eosinophils in blood and nasal polyps of severe asthma patients.

In mice models, eosinophils have been divided into different subpopulations with distinct phenotypes and functions, based on CD62L and CD101 patterns of membrane expression. Limited data are available in humans. To investigate eosinophils subpopulations in peripheral blood (PB) and nasal polyp tissue (NP) from severe eosinophilic asthma (SEA) patients plus concomitant chronic rhinosinusitis with nasal polyps (CRSwNP). We recruited 23 SEA patients (14 with CRSwNP); as controls, we enrolled 15 non-severe asthma patients, 15 allergic rhinitis patients without asthma and 15healthy donors. Eosinophils were isolated from PB and NP and analysed by FACS. Eotaxin-3 and eotaxin-1mRNA expression in NP tissue was also evaluated. A significantly higher percentage of circulating CD62Llow cells was observed in SEA, as compared with controls, expressing higher levels of CCR3, CD69 and lower levels of CD125 (IL-5R), CRTH2, CD86 and CD28 in comparison with CD62Lbright cells. In NP, eosinophils showed a high proportion of CD62Llow phenotype, significantly greater than that observed in PB. Surface expression of IL-3R, IL-5R, CD69 and CD86 was significantly higher in CD62Llow eosinophils from NP than in those from blood. Moreover, eotaxin-3mRNA expression positively correlated with the percentage of CD62Llow cells in NP. Two different eosinophil subphenotypes can be identified in blood and NP of SEA patients, with a preferential accumulation of CD62Llow inflammatory cells in NP.

Awareness of asthma and its management in primary school teachers in Baghdad, Iraq.

Background: Asthma is a major global health issue characterized by chronic airway inflammation. It is linked to a high disease burden and disproportionately high healthcare utilization in severe, uncontrolled cases compared to non-severe asthma. We aimed to conduct this survey among primary school teachers in Baghdad, Iraq, to assess their level of knowledge about asthma and confidence in managing school children with asthma. Methods: This was a prospective cross-sectional study conducted in the Karkh and Rusafa areas of Baghdad. The study adopted a questionnaire for assessing the asthma knowledge and confidence scores regarding the management of asthmatic children. This questionnaire contained 29-multiple true-false questions based on different aspects, including facts about asthma and the management of asthma by teachers. Results: The questionnaire for testing teachers' knowledge of asthma and confidence scores regarding the management of asthmatic children was distributed to 150 teachers. 103 (68%) teachers completed the questionnaire fully. Approximately 71% of teachers answered the question 'What are the three main symptoms of asthma?' correctly i.e., answering either one symptom (35.9%) or two symptoms (35.9%) correctly. A relatively smaller number of teachers (16.5%) mentioned all three symptoms correctly. Across the 29-multiple true-false questions, more than 75% of teachers answered 11 questions correctly, 50-74% of teachers answered the rest of eight questions appropriately, and <50% of teachers answered the remaining 10 questions properly. The statistical evaluation indicated that the mean total knowledge score about asthma for all the teachers was 20.27 ± 2.97 and the mean total confidence score regarding the management of asthmatic children was 72.44 ± 13.61. Conclusions: This study suggests that teachers from the schools in Karkh and Rusafa areas of Baghdad appear to be self-confident in their ability and knowledge to help and manage children with asthma.

Nrf2 regulates downstream genes by targeting miR-29b in severe asthma and the role of grape seed proanthocyanidin extract in a murine model of steroid-insensitive asthma

Context Grape seed proanthocyanidin extract (GSPE) is effective in treating severe asthma (SA). Objective To examine the relationship between Nrf2-miR-29b axis and SA, and to detect whether preventive use of GSPE relieves SA via it. Materials and methods We recruited 10 healthy controls, 10 patients with non-severe asthma (nSA), and 9 patients with SA from February 2017 to December 2017. Peripheral blood mononuclear cells from these volunteers were extracted. A murine model of steroid-insensitive asthma was established in six-week-old female BALB/c mice that were sensitised and challenged with OVA, Al(OH)3 and LPS for 31 days. Mice in the treated groups were injected with DXM (5 mg/kg/d), with or without GSPE (100 mg/kg/d). Control group received PBS. We performed quantitative real-time PCR, western blot and luciferase reporter assay in animal and cell models. Results SA group demonstrated significantly lower concentrations of Nrf2 protein, Nrf2 mRNA, and miR-29b than nSA group and control group. Conversely, higher levels of platelet derived growth factor C (PDGFC), phosphoinositide-3-kinase regulatory subunit 1 (PIK3R1), and collagen type III alpha 1 (COL3A1) were measured in SA than in the other two groups. PDGFC, PIK3R1, and COL3A1 were the target genes of miR-29b. GSPE + DXM significantly elevated the expression of Nrf2 (+188%), Nrf2 mRNA (+506%), and miR-29b (+201%), and significantly reduced the expression of PDGFC (−72%), PIK3R1 (−40%), and COL3A1 (−65%) compared with OVA + LPS. Conclusions Nrf2-miR-29b axis is involved in the pathogenesis of SA. GSPE, as an adjuvant drug, maybe a potential therapeutic agent for SA.

Phenotypic clusters on computed tomography reflects asthma heterogeneity and severity.

BackgroundAsthma is a heterogeneous inflammatory airway disorder with various phenotypes. Quantitative computed tomography (QCT) methods can differentiate among lung diseases through accurate assessment of the location, extent, and severity of the disease. The purpose of this study was to identify asthma clusters using QCT metrics of airway and parenchymal structure, and to identify associations with visual analyses conducted by radiologists.MethodsThis prospective study used input from QCT-based metrics including hydraulic diameter (Dh), luminal wall thickness (WT), functional small airway disease (fSAD), and emphysematous lung (Emph) to perform a cluster analysis and made comparisons with the visual grouping analysis conducted by radiologists based on site of airway involvement and remodeling evaluated.ResultsA total of 61 asthmatics of varying severities were grouped into 4 clusters. From C1 to C4, more severe lung function deterioration, higher fixed obstruction rate, and more frequent asthma exacerbations were observed in the 5-year follow-up period. C1 presented non-severe asthma with increased WT, Dh of proximal airways, and fSAD. C2 was mixed with non-severe and severe asthmatics, and showed bronchodilator responses limited to the proximal airways. C3 and C4 included severe asthmatics that showed a reduced Dh of the proximal airway and diminished bronchodilator responses. While C3 was characterized by severe allergic asthma without fSAD, C4 included ex-smokers with high fSAD% and Emph%. These clusters correlated well with the grouping done by radiologists and clinical outcomes.ConclusionsFour QCT imaging-based clusters with distinct structural and functional changes in proximal and small airways can stratify heterogeneous asthmatics and can be a complementary tool to predict clinical outcomes.

Epithelial Autoantigen-Specific IgG Antibody Enhances Eosinophil Extracellular Trap Formation in Severe Asthma.

PurposeThere have been autoimmune mechanisms for the pathogenesis of severe asthma (SA) involving epithelial autoantigen-specific antibodies. This study aimed to find the function of these antibodies in the formation of eosinophil extracellular traps (EETs), contributing to the development of SA.MethodsPatients with SA (n = 11), those with patients with nonsevere asthma (NSA, n = 41), and healthy controls (HCs, n = 26) were recruited to evaluate levels of epithelial antigens and autoantigen-specific antibodies. Moreover, the significance of epithelial autoantigen-specific antibodies in association with EET production was investigated ex vivo and in vivo.ResultsSignificantly higher levels of serum cytokeratin (CK) 18 and CK18-specific IgG were observed in patients with SA than in those with NSA (P = 0.001 and P = 0.031, respectively), while no differences were found in serum CK19 or CK19-specific immunoglobulin G (IgG). Moreover, levels of serum CK18 were positively correlated with total eosinophil counts (r = 0.276, P = 0.048) in asthmatics, while a negative correlation was noted between levels of serum CK18 and forced expiratory volume in 1 second (FEV1) %. In the presence of CK18-specific IgG, peripheral eosinophils from asthmatics released EETs, which further increased CK18 production from airway epithelial cells. In severe asthmatic mice, CK18 expression and CK18-specific IgG production were enhanced in the lungs, where EET treatment enhanced CK18 expression and CK18-specific IgG production, either of which was not suppressed by dexamethasone.ConclusionsThese suggest that EETs could enhance epithelial autoantigen (CK18)-induced autoimmune responses, further stimulating EET production and type 2 airway responses, which is a new therapeutic target for SA.

Peripheral blood transcriptomic clusters uncovered immune phenotypes of asthma

BackgroundTranscriptomic analysis has been used to elucidate the complex pathogenesis of heterogeneous disease and may also contribute to identify potential therapeutic targets by delineating the hub genes. This study aimed to investigate whether blood transcriptomic clustering can distinguish clinical and immune phenotypes of asthmatics, and microbiome in asthmatics.MethodsTranscriptomic expression of peripheral blood mononuclear cells (PBMCs) from 47 asthmatics and 21 non-asthmatics was measured using RNA sequencing. A hierarchical clustering algorithm was used to classify asthmatics. Differentially expressed genes, clinical phenotypes, immune phenotypes, and microbiome of each transcriptomic cluster were assessed.ResultsIn asthmatics, three distinct transcriptomic clusters with numerously different transcriptomic expressions were identified. The proportion of severe asthmatics was highest in cluster 3 as 73.3%, followed by cluster 2 (45.5%) and cluster 1 (28.6%). While cluster 1 represented clinically non-severe T2 asthma, cluster 3 tended to include severe non-T2 asthma. Cluster 2 had features of both T2 and non-T2 asthmatics characterized by the highest serum IgE level and neutrophil-dominant sputum cell population. Compared to non-asthmatics, cluster 1 showed higher CCL23 and IL1RL1 expression while the expression of TREML4 was suppressed in cluster 3. CTSD and ALDH2 showed a significant positive linear relationship across three clusters in the order of cluster 1 to 3. No significant differences in the diversities of lung and gut microbiomes were observed among transcriptomic clusters of asthmatics and non-asthmatics. However, our study has limitations in that small sample size data were analyzed with unmeasured confounding factors and causal relationships or function pathways were not verified.ConclusionsGenetic clustering based on the blood transcriptome may provide novel immunological insight, which can be biomarkers of asthma immune phenotypes.Trial registration Retrospectively registered