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Analysis of Pulmonary Dendritic Cell Maturation and Migration during Allergic Airway Inflammation
Published on: July 23, 2012
Respiratory tract dendritic cells in paediatric asthma
R Brugha1, N Mushtaq, N E McCarthy
1Asthma UK Centre for Applied Research, Centre for Paediatrics, Blizard Institute, Queen Mary, University of London, London, UK.
Insights
Children with asthma have more airway dendritic cells (DC) with reduced CD86 expression, indicating altered cell function. This study characterizes DC in paediatric asthma using sputum analysis.
Area of Science:
- Immunology
- Pulmonology
- Pediatrics
Background:
- Airway dendritic cells (DC) are key players in lung inflammation during asthma.
- Characteristics of DC in healthy children versus those with asthma are not well understood.
Purpose of the Study:
- To identify changes in DC subset distribution and activation in pediatric asthma.
- To analyze induced sputum samples from healthy and asthmatic children using flow cytometry.
Main Methods:
- Spirometry and skin prick tests assessed lung function and atopy.
- Seven-color flow cytometry analyzed induced sputum for airway DC populations.
- DC were identified as lineage(-) HLA-DR(+) cells expressing CD11c (conventional) or CD123 (plasmacytoid).
Main Results:
- DC were found in all sputum samples, with a higher proportion in asthmatic children (1.29% vs. 0.67%).
- Asthmatic children showed significantly reduced CD86 expression on DC (59.7% vs. 73.4%).
- CD1c(+) cells were abundant (57% of DC), with low frequencies of CD141 or CD123 expressing cells.
Conclusions:
- Airway DC can be non-invasively characterized in pediatric sputum using flow cytometry.
- Children with asthma have increased airway DC with reduced CD86 expression.
- Findings suggest altered DC trafficking or maturation in pediatric asthma, potentially influenced by steroid therapy.
Background:
Airway dendritic cells (DC) are critical mediators of lung inflammation in asthma, but the characteristics of DC in the airways of healthy children, and children with asthma, are currently unknown.
Objective:
We sought to identify changes in DC subset distribution and activation profile in paediatric asthma using flow cytometry to analyse induced sputum samples obtained from healthy and asthmatic children.
Methods:
Lung function and atopic status were determined by spirometry and skin prick testing. Induced sputum samples were analysed using 7-colour flow cytometry to identify airway DC populations (lineage(-) HLA-DR(+) sputum cells expressing either CD11c as conventional DC or CD123 as plasmacytoid DC).
Results:
Sputum samples containing lower airway plugs were obtained from 10 healthy children and 8 children with asthma. Lineage(-) HLA-DR(+) DC were successfully identified in all samples, and DC comprised a significantly higher proportion of sputum cells in children with asthma compared with age-matched healthy controls (1.29% vs. 0.67%, P = 0.02). DC expression of the costimulatory marker CD86 was significantly reduced in asthmatic children (73.4% vs. 59.7%, P = 0.04). Sputum DC also included numerous CD1c(+) cells (mean 57% of the total DC population) and low frequencies of cells expressing the subset markers CD141 or CD123, although the proportions of these did not differ between groups.
Conclusions:
Airway DC can be identified and characterized non-invasively using flow cytometry to analyse paediatric sputum samples. Our data reveal that children with steroid-treated asthma exhibit increased frequency of airway DC with reduced expression of the costimulatory marker CD86, suggesting altered trafficking and/or maturation of these cells either due to asthma or steroid therapies.
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