scholarly journals The usefulness of competitive PCR: airway gene expression of IL-5, IL-4, IL-4δ2, IL-2, and IFNγ in asthma

Thorax ◽  
2001 ◽  
Vol 56 (7) ◽  
pp. 541-548
Author(s):  
E M Glare ◽  
M Divjak ◽  
M J Bailey ◽  
E H Walters

BACKGROUNDAsthma has been described as an eosinophilic bronchitis driven by interleukin(IL)-4 and IL-5. The quantification of cytokine mRNA levels in airway samples has been confounded by housekeeping gene expression which differs between and within asthmatics and controls.METHODSThe usefulness of competitive reverse transcriptase-polymerase chain reaction (RT-PCR) that is independent of housekeeping gene expression for quantitating the mRNA for interferon (IFN)γ, IL-2, IL-5, IL-4 and its receptor antagonist encoding splicing variant IL-4δ2 was determined in a cross sectional study of 45 normal control subjects and 111 with asthma.RESULTSAtopic controls and atopic asthmatic subjects expressed more IL-5 than non-atopic controls (p<0.02) in bronchoalveolar lavage (BAL) cells, but not in biopsy specimens. IL-5 mRNA expression in BAL cells from asthmatic subjects using inhaled corticosteroids (ICS) was significantly lower than those not receiving ICS (p=0.04). IL-2 mRNA levels differed with steroid use in biopsy specimens but not in BAL cells. IFNγ, IL-4, and IL-4δ2 mRNA levels did not differ between any groups and were not affected by steroid use. IL-4 and IL-4δ2 mRNA levels were positively correlated (p<0.0001), suggesting coordinated transcription.CONCLUSIONSWhile the signal differentiation of competitive PCR in asthma may rival that of in situ hybridisation and immunohistochemistry, the method is expensive and wasteful of material.

Nutrients ◽  
2019 ◽  
Vol 12 (1) ◽  
pp. 74 ◽  
Author(s):  
Isobel Stoodley ◽  
Manohar Garg ◽  
Hayley Scott ◽  
Lesley Macdonald-Wicks ◽  
Bronwyn Berthon ◽  
...  

Asthma is a chronic inflammatory airway disease, associated with systemic inflammation. Omega-3 polyunsaturated fatty acids (n-3 PUFA) have established anti-inflammatory effects, thus having potential as an adjunct therapy in asthma. This study aimed to compare erythrocyte n-3 PUFA in adults with (n = 255) and without (n = 137) asthma and determine the relationship between erythrocyte n-3 PUFA and clinical asthma outcomes. Subjects had blood collected, lung function measured and Juniper Asthma Control Questionnaire (ACQ) score calculated. Fatty acids were measured in erythrocyte membranes by gas chromatography, and the omega-3 index (O3I) was calculated (% eicosapentaenoic acid + % docosahexaenoic acid). O3I was similar in subjects with and without asthma (p = 0.089). A higher O3I was observed in subjects with controlled or partially controlled asthma (ACQ < 1.5) compared to subjects with uncontrolled asthma (ACQ ≥ 1.5) (6.0% (5.4–7.2) versus 5.6% (4.6–6.4) p = 0.033). Subjects with a high O3I (≥8%) had a lower maintenance dose of inhaled corticosteroids (ICS) compared to those with a low O3I (<8%) (1000 μg (400–1000) versus 1000 μg (500–2000) p = 0.019). This study demonstrates that a higher O3I is associated with better asthma control and with lower ICS dose, suggesting that a higher erythrocyte n-3 PUFA level may have a role in asthma management.


2013 ◽  
Vol 12 (1) ◽  
Author(s):  
Annie Bouchard-Mercier ◽  
Ann-Marie Paradis ◽  
Iwona Rudkowska ◽  
Simone Lemieux ◽  
Patrick Couture ◽  
...  

2018 ◽  
Vol 4 (3) ◽  
pp. 00015-2018 ◽  
Author(s):  
Moana Mika ◽  
Izabela Nita ◽  
Laura Morf ◽  
Weihong Qi ◽  
Seraina Beyeler ◽  
...  

Compartmentalisation of the respiratory tract microbiota in patients with different chronic obstructive pulmonary disease (COPD) severity degrees needs to be systematically investigated. In addition, it is unknown if the inflammatory and emphysematous milieux in patients with COPD are associated with changes in the respiratory tract microbiota and host macrophage gene expression.We performed a cross-sectional study to compare non-COPD controls (n=10) to COPD patients (n=32) with different disease severity degrees. Samples (n=187) were obtained from different sites of the upper and lower respiratory tract. Microbiota analyses were performed by 16S ribosomal RNA gene sequencing and host gene expression analyses by quantitative real-time PCR of distinct markers of bronchoalveolar lavage cells.Overall, the microbial communities of severe COPD (Global Initiative for Chronic Obstructive Lung Disease (GOLD) grade 3/4) patients clustered significantly differently to controls and less severe COPD (GOLD 1/2) patients (permutational multivariate ANOVA (MANOVA), p=0.001). However, we could not detect significant associations between the different sampling sites in the lower airways. In addition, the chosen set of host gene expression markers significantly separated COPD GOLD 3/4 patients, and we found correlations between the composition of the microbiota and the host data.In conclusion, this study demonstrates associations between host gene expression and microbiota profiles that may influence the course of COPD.


2015 ◽  
Vol 35 (9) ◽  
pp. 569-574
Author(s):  
Bernardino Alcázar Navarrete ◽  
Pedro José Romero Palacios ◽  
Antonio Aguilar-Salvatierra ◽  
Javier Guardia ◽  
Gerardo Gómez-Moreno

2021 ◽  
Vol 4 ◽  
pp. 13-14
Author(s):  
John C. Su ◽  
Nikolay Murashkin ◽  
Andreas Wollenberg ◽  
Åke Svensson ◽  
Pavel Chernyshov ◽  
...  

2021 ◽  
Author(s):  
Renan Paulo Martin ◽  
Patricia Varela ◽  
Caio Peres Gomes ◽  
Maryana Mara Marins ◽  
Rafael Filippelli-Silva ◽  
...  

Abstract Skin aging involves genetic, environmental and hormonal factors. Facial wrinkles also depend on muscular activity. Gene expression investigation may be useful for new anti-aging products. To evaluate structure and gene expression differences among exposed and unexposed skin in menopausal women. Cross-sectional study, including 15 menopausal women, 55-65yo, phototype III; photo-exposed, periorbital wrinkles (A1), preauricular, not wrinkled (A2), and unexposed gluteal (A3) areas were described and compared by non-invasive measures, histology, immunohistochemistry and gene expression (RNASeq); participants mean age was 61yo, presenting moderate periorbital wrinkles and light facial photodamage. Higher roughness, wrinkles number and echogenicity were observed in A1 and A2 versus A3. Decreased epidermal thickness and dermal collagen IV were demonstrated in A1 versus A2 and A3. Exposed areas impacted different pathways compared to unexposed. Exposed wrinkled skin (A1) showed impact on cell movement with decreased inflammatory activation state. Pathways related to lipid and aminoacids metabolism were modulated in non-wrinkled exposed (A2) compared to unexposed (A3) skin. Expected histological findings and gene expression differences among areas were observed. Photoaging in menopausal women may modulate lipid and aminoacids metabolism and decrease inflammatory and keratinization pathways, cellular homeostasis, immune response, fibrogenesis and filament formation. These findings may help development of new therapies for skin health and aging control.


2020 ◽  
Author(s):  
Monica Zavala-Solares ◽  
Gabriela Fonseca-Camarillo ◽  
Miguel Valdovinos ◽  
Julio Granados ◽  
Guido Grajales-Figueroa ◽  
...  

Abstract Background: Patients clinical endoscopic phenotypes in gastroesophageal reflux disease (GERD) are classified as: Barrett's esophagus (BE), erosive esophagitis (EE) and non-erosive gastroesophageal reflux disease (NERD). NERD are subclassified in Abnormal acid exposure (AAE) and Normal acid exposure (NAE) according to pH monitoring study. The aim of this study was to characterize genes involved in the pathophysiology and immune response of GERD.Methods: This is an observational and cross-sectional study. All patients with BE, EE, AAE, NAE and control group were subjected to a superior endoscopy (with biopsies of esophageal mucosa). The cytokine mRNA relative quantification of target genes was conducted by RT-PCR. Changes in gene expression were assessed of the genes associated with inflammation in each disease phenotype. Statistical analysis of differential gene expression was performed by using Dunn's Multiple Comparison non-parametric test. A p value < 0.05 was considered as significant. Results: A total of 82 patients were included and they were divided into the following groups: Group BE 16 (19.51%), Group EE 23 (28.04%), Group AAE 13 (15.86%), NAE (15.86%) and Control Group 17 (20.73%). When comparing with control group we found: patients with BE showed an increased expression of IL-8 (P<0.005) and higher levels of: IL-10 and MMP-3, MMP-9 as well; patients with EE had higher levels of IL-1B, IL-6 and IL-10 (P<0.005), patients with AAE showed an increased expression of Il-1B, Il-6, IFN-γ and TNF-α (P<0.005). AAE had a higher expression of Il-1B and TNF-α than NAE (P<0.005). Conclusions: This study demonstrates the differential expression of mediators of inflammation in the esophageal mucosa of patients in GERD endoscopic phenotypes. MMP3 could be implicated in damage to esophageal mucosa. IL-1B and TNF-α could be a differential diagnosis between AAE and NAE in the non-erosive phenotype from endoscopic biopsies.


Thorax ◽  
2020 ◽  
Vol 75 (10) ◽  
pp. 835-841
Author(s):  
Luke Evan Barry ◽  
Ciaran O’Neill ◽  
Liam G Heaney

BackgroundAllostatic load, a measure of early ageing or ‘wear and tear’ from adapting to environmental challenges, has been suggested as a framework with which to understand the stress-related disruption of multiple biological systems which may be linked to asthma. Considering the socioeconomic context is also critical given asthma and allostatic overload are more common in lower socioeconomic groups.AimsEstimate the relationship between allostatic load and its constituent biomarkers, asthma and corticosteroid prescribing while controlling for socioeconomic status.MethodsData from Understanding Society (a nationally representative survey of UK community-dwelling adults) waves 1–3 (2009–2012) allowed the identification of a sex-specific risk profile across 12 biomarkers used to construct an Allostatic Load Index for a sample of 9816 adults. Regression analyses were used to examine the association of asthma status and corticosteroid prescriptions with allostatic load and its constituent biomarkers while controlling for socioeconomic status (n=9805).ResultsSubjects with currently treated asthma and no corticosteroid prescription have an allostatic load 1.21 times higher than those without asthma (p<0.001). Asthmatic subjects in receipt of inhaled corticosteroids had an allostatic load, approximately 1.12 times higher than those without asthma (p<0.001). This association persisted in sensitivity analyses and appeared to be driven by an association with specific biomarkers (dehydroepiandrosterone-sulfate, waist-to-height ratio and C-reactive protein).ConclusionEarly ageing, in the form of a higher allostatic load, was present even in the mildest asthma group not receiving inhaled corticosteroids. Allostatic load is helpful in understanding the increased all-cause mortality and multimorbidity observed in asthma.


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