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An In vitro Model to Study Immune Responses of Human Peripheral Blood Mononuclear Cells to Human Respiratory Syncytial Virus Infection
Published on: December 10, 2013
Interferon response to respiratory syncytial virus by bronchial epithelium from children with asthma is inversely
Matthew C Altman1, Stephen R Reeves2, Andrew R Parker3
1Division of Allergy and Infectious Diseases, Department of Medicine, University of Washington, Seattle, Wash; Benaroya Research Institute, Seattle, Wash.
Insights
Respiratory syncytial virus (RSV) infection in children with asthma triggers distinct gene expression in airway cells. This exaggerated interferon response correlates with airway obstruction, suggesting a mechanism for lung function decline.
Area of Science:
- Pediatric Pulmonology
- Immunology
- Respiratory Medicine
Background:
- Childhood respiratory viral infections, including RSV, are linked to asthma development.
- Understanding the cellular response to RSV in asthmatic children is crucial for identifying disease mechanisms.
Purpose of the Study:
- To investigate gene expression patterns in bronchial epithelial cells (BECs) from children with asthma following RSV infection.
- To determine if these patterns correlate with lung function in the BEC donors.
Main Methods:
- Primary BECs from children with and without asthma were cultured at an air-liquid interface and infected with RSV.
- Gene expression was analyzed using RNA sequencing, and lung function was assessed via spirometry.
Main Results:
- RSV infection significantly increased the expression of 6199 genes in BECs from children with asthma.
- A subset of 195 genes, enriched for viral response and interferon pathways, showed significantly higher expression in asthmatic children with airway obstruction compared to those without obstruction or healthy controls.
Conclusions:
- BECs from asthmatic children with obstructive physiology exhibit an amplified type I and III interferon response to RSV infection.
- This exaggerated interferon response may contribute to lung function decline in a subset of pediatric asthma patients.
Background:
Respiratory viral infection in early childhood, including that from respiratory syncytial virus (RSV), has been previously associated with the development of asthma.
Objective:
We aimed to determine whether ex vivo RSV infection of bronchial epithelial cells (BECs) from children with asthma would induce specific gene expression patterns and whether such patterns were associated with lung function among BEC donors.
Methods:
Primary BECs from carefully characterized children with asthma (n = 18) and matched healthy children without asthma (n = 8) were differentiated at an air-liquid interface for 21 days. Air-liquid interface cultures were infected with RSV for 96 hours and RNA was subsequently isolated from BECs. In each case, we analyzed gene expression using RNA sequencing and assessed differences between conditions by linear modeling of the data. BEC donors completed spirometry to measure lung function.
Results:
RSV infection of BECs from subjects with asthma, compared with uninfected BECs from subjects with asthma, led to a significant increase in expression of 6199 genes. There was significantly greater expression of 195 genes in BECs from children with asthma and airway obstruction (FEV1/forced vital capacity < 0.85 and FEV1 < 100% predicted) than in BECs from children with asthma without obstruction, or in BECs from healthy children. These specific genes were found to be highly enriched for viral response genes induced in parallel with types I and III interferons.
Conclusions:
BECs from children with asthma and with obstructive physiology exhibit greater expression of types I and III interferons and interferon-stimulated genes than do cells from children with normal lung function, and expression of interferon-associated genes correlates with the degree of airway obstruction. These findings suggest that an exaggerated interferon response to viral infection by airway epithelial cells may be a mechanism leading to lung function decline in a subset of children with asthma.
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