Abstract
The choice of model used to study human respiratory syncytial virus (RSV) infection is extremely important. RSV is a human pathogen that is exquisitely adapted to infection of human hosts. Rodent models, such as mice and cotton rats, are semi-permissive to RSV infection and do not faithfully reproduce hallmarks of RSV disease in humans. Furthermore, immortalized airway-derived cell lines, such as HEp-2, BEAS-2B, and A549 cells, are poorly representative of the complexity of the respiratory epithelium. The development of a well-differentiated primary pediatric airway epithelial cell models (WD-PAECs) allows us to simulate several hallmarks of RSV infection of infant airways. They therefore represent important additions to RSV pathogenesis modeling in human-relevant tissues. The following protocols describe how to culture and differentiate both bronchial and nasal primary pediatric airway epithelial cells and how to use these cultures to study RSV cytopathogenesis.
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References
Zhang L, Peeples ME, Boucher RC et al (2002) Respiratory syncytial virus infection of human airway epithelial cells is polarized, specific to ciliated cells, and without obvious cytopathology. J Virol 76:5654–5666. doi:10.1128/JVI.76.11.5654-5666.2002
Villenave R, Shields MD, Power UF (2013) Respiratory syncytial virus interaction with human airway epithelium. Trends Microbiol 21:238–244. doi:10.1016/j.tim.2013.02.004
Gray TE, Guzman K, Davis CW et al (1996) Mucociliary differentiation of serially passaged normal human tracheobronchial epithelial cells. Am J Respir Cell Mol Biol 14:104–112. doi:10.1165/ajrcmb.14.1.8534481
Villenave R, Thavagnanam S, Sarlang S et al (2012) In vitro modeling of respiratory syncytial virus infection of pediatric bronchial epithelium, the primary target of infection in vivo. Proc Natl Acad Sci 109:5040–5045. doi:10.1073/pnas.1110203109
Pezzulo AA, Starner TD, Scheetz TE et al (2011) The air-liquid interface and use of primary cell cultures are important to recapitulate the transcriptional profile of in vivo airway epithelia. Am J Physiol Lung Cell Mol Physiol 300:L25–L31. doi:10.1152/ajplung.00256.2010
Guo-Parke H, Canning P, Douglas I et al (2013) Relative respiratory syncytial virus cytopathogenesis in upper and lower respiratory tract epithelium. Am J Respir Crit Care Med 188:842–851. doi:10.1164/rccm.201304-0750OC
Tripp RA (2013) Modeling respiratory syncytial virus cytopathogenesis in the human airway. Am J Respir Crit Care Med 188:766–767. doi:10.1164/rccm.201308-1491ED
Doherty GM, Christie SN, Skibinski G et al (2003) Non-bronchoscopic sampling and culture of bronchial epithelial cells in children. Clin Exp Allergy 33:1221–1225. doi:10.1046/j.1365-2222.2003.01752.x
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Broadbent, L., Villenave, R., Guo-Parke, H., Douglas, I., Shields, M.D., Power, U.F. (2016). In Vitro Modeling of RSV Infection and Cytopathogenesis in Well-Differentiated Human Primary Airway Epithelial Cells (WD-PAECs). In: Tripp, R., Jorquera, P. (eds) Human Respiratory Syncytial Virus. Methods in Molecular Biology, vol 1442. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3687-8_9
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DOI: https://doi.org/10.1007/978-1-4939-3687-8_9
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