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Published on: March 10, 2016
Comparison of techniques for obtaining lower airway epithelial cells from children
P S McNamara1, A Kicic, E N Sutanto
1Institute of Child Health, Royal Liverpool Children's Hospital; Eaton Rd, Liverpool, L12 2AP, UK. mcnamp@liv.ac.uk
Insights
A new bronchoscope-directed method for collecting airway epithelial cells (AECs) in children yields more cells than the traditional blind technique. This improved method is well-tolerated and valuable for childhood asthma research.
Area of Science:
- Pediatric Pulmonology
- Cell Biology
- Respiratory Medicine
Background:
- Airway epithelial cells (AECs) are crucial in asthma, acting as the initial defense against pathogens and allergens.
- Traditional methods for collecting pediatric AECs, like blind endotracheal brushing, are becoming less applicable due to increased laryngeal mask use.
- A novel portable bronchoscope-directed technique offers an alternative for AEC sampling in children.
Purpose of the Study:
- To compare the efficacy and safety of bronchoscope-directed versus blind brushing techniques for collecting AECs in children.
- To evaluate cellular yield, cell lineage, and cytokine profiles from both sampling methods.
- To assess the suitability of these techniques for research in pediatric respiratory diseases, including asthma.
Main Methods:
- 81 children undergoing elective surgery were recruited and categorized by atopic status and respiratory symptoms.
- AEC samples were collected using both blind nonbronchoscopic and portable bronchoscope-directed brushing techniques.
- Collected cells underwent immunocytochemistry for lineage confirmation, and cultures were analyzed for cytokine concentrations.
Main Results:
- Both sampling techniques were well-tolerated, with mild cough reported in 10% of subjects.
- The bronchoscope-directed technique yielded a significantly higher mean cell count (5.1±2.4 x 10^6 cells) compared to blind brushing (3.1±1.4 x 10^6 cells).
- Immunocytochemistry confirmed epithelial cell lineage, and cytokine profiles in culture supernatants were consistent across both methods and between asthmatic and nonasthmatic children.
Conclusions:
- Portable bronchoscope-directed brushing is a well-tolerated, effective method for obtaining higher yields of AECs in children compared to blind brushing.
- This technique preserves cellular characteristics and cytokine patterns, making it a valuable tool for pediatric respiratory disease research, particularly for childhood asthma.
- The bronchoscope-directed approach offers a quick and useful alternative for AEC collection when laryngeal mask use limits traditional methods.
Abstract:
Airway epithelial cells (AECs) are important in asthma as they are the first cells to encounter pathogens/allergens. In children, AECs can be obtained using a "blind" nonbronchoscopic technique through an endotracheal tube. However, due to the increasing use of laryngeal masks the number of children in whom this technique is applicable has become limited. Recently, the present authors began to use a portable "bronchoscope-directed" technique to sample AECs. The current study compares both techniques in both asthmatic and nonasthmatic children. A total of 81 children undergoing elective surgery, were grouped according to atopic status and respiratory symptoms. Cellular yield of blind and bronchoscope-directed brushings were compared and immunocytochemistry performed. AECs were cultured and cytokine analysis of culture supernatant undertaken. Both techniques were equally well-tolerated, with the only adverse effect being a cough in 10% of the subjects. The mean+/-SD cell yield was higher in bronchoscope-directed than blind brushings (5.1+/-2.4 versus 3.1+/-1.4x10(6) cells). Immunocytochemistry confirmed an epithelial cell lineage. Culture supernatant cytokine concentrations were similar regardless of sampling technique with patterns preserved between asthmatic and healthy nonatopic phenotypes. Compared with blind brushing portable bronchoscope-directed brushing is well-tolerated, yields significantly more cells and is a potentially quick and useful technique for obtaining airway epithelial cells for research into childhood respiratory disease, specifically asthma.
