Human Airway Epithelial Cell Cultures for Modeling Respiratory Syncytial Virus Infection



Respiratory syncytial virus (RSV) is an important human respiratory pathogen with narrow species tropism. Limited availability of human pathologic specimens during early RSV-induced lung disease and ethical restrictions for RSV challenge studies in the lower airways of human volunteers has slowed our understanding of how RSV causes airway disease and greatly limited the development of therapeutic strategies for reducing RSV disease burden. Our current knowledge of RSV infection and pathology is largely based on in vitro studies using nonpolarized epithelial cell-lines grown on plastic or in vivo studies using animal models semipermissive for RSV infection. Although these models have revealed important aspects of RSV infection, replication, and associated inflammatory responses, these models do not broadly recapitulate the early interactions and potential consequences of RSV infection of the human columnar airway epithelium in vivo. In this chapter, the pro et contra of in vitro models of human columnar airway epithelium and their usefulness in respiratory virus pathogenesis and vaccine development studies will be discussed. The use of such culture models to predict characteristics of RSV infection and the correlation of these findings to the human in vivo situation will likely accelerate our understanding of RSV pathogenesis potentially identifying novel strategies for limiting the severity of RSV-associated airway disease.


  1. Bartlett EJ, Cruz AM, Esker J, Castano A, Schomacker H, Surman SR, Hennessey M, Boonyaratanakornkit J, Pickles RJ, Collins PL, Murphy BR, Schmidt AC (2008a) Human parainfluenza virus type 1 C proteins are nonessential proteins that inhibit the host interferon and apoptotic responses and are required for efficient replication in nonhuman primates. J Virol 82(18):8965–8977PubMedCentralPubMedCrossRefGoogle Scholar
  2. Bartlett EJ, Hennessey M, Skiadopoulos MH, Schmidt AC, Collins PL, Murphy BR, Pickles RJ (2008b) Role of interferon in the replication of human parainfluenza virus type 1 wild type and mutant viruses in human ciliated airway epithelium. J Virol 82(16):8059–8070PubMedCentralPubMedCrossRefGoogle Scholar
  3. Bove PF, Grubb BR, Okada SF, Ribeiro CM, Rogers TD, Randell SH, O’Neal WK, Boucher RC (2011) Human alveolar type II cells secrete and absorb liquid in response to local nucleotide signaling. J Biol Chem 285(45):34939–34949Google Scholar
  4. Button B, Cai LH, Ehre C, Kesimer M, Hill DB, Sheehan JK, Boucher RC, Rubinstein M (2012) A periciliary brush promotes the lung health by separating the mucus layer from airway epithelia. Science 337(6097):937–941Google Scholar
  5. Collins PL (1990) O glycosylation of glycoprotein G of human respiratory syncytial virus is specified within the divergent ectodomain. J Virol 64(8):4007–4012PubMedCentralPubMedGoogle Scholar
  6. Fulcher ML, Gabriel S, Burns KA, Yankaskas JR, Randell SH (2005) Well-differentiated human airway epithelial cell cultures. Methods Mol Med 107:183–206PubMedGoogle Scholar
  7. Fulcher ML, Gabriel SE, Olsen JC, Tatreau JR, Gentzsch M, Livanos E, Saavedra MT, Salmon P, Randell SH (2009) Novel human bronchial epithelial cell lines for cystic fibrosis research. Am J Physiol Lung Cell Mol Physiol 296(1):L82–L91PubMedCrossRefGoogle Scholar
  8. Harkema J, Mariassy A, St. George J, Hyde DM, Plopper CG (1994) Epithelial cells of the conducting airways: a species comparison. In: Farmer SG, Hay DWP (eds) The airway epithelium, vol 55. Lung Biology in Health and Disease. Marcel Dekker, New York, pp 3–39Google Scholar
  9. Ilyushina NA, Ikizler MR, Kawaoka Y, Rudenko LG, Treanor JJ, Subbarao K, Wright PF (2012) Comparative study of influenza virus replication in MDCK cells and in primary cells derived from adenoids and airway epithelium. J VirolGoogle Scholar
  10. Johnson JE, Gonzales RA, Olson SJ, Wright PF, Graham BS (2007) The histopathology of fatal untreated human respiratory syncytial virus infection. Mod Pathol 20(1):108–119PubMedCrossRefGoogle Scholar
  11. Karron RA, Buonagurio DA, Georgiu AF, Whitehead SS, Adamus JE, Clements-Mann ML, Harris DO, Randolph VB, Udem SA, Murphy BR, Sidhu MS (1997) Respiratory syncytial virus (RSV) SH and G proteins are not essential for viral replication in vitro: clinical evaluation and molecular characterization of a cold-passaged, attenuated RSV subgroup B mutant. Proc Natl Acad Sci USA 94(25):13961–13966PubMedCrossRefGoogle Scholar
  12. Kesimer M, Ehre C, Burns KA, Davis CW, Sheehan JK, Pickles RJ (2012) Molecular organization of the mucins and glycocalyx underlying mucus transport over mucosal surfaces of the airways. Mucosal ImmunolGoogle Scholar
  13. Kesimer M, Kirkham S, Pickles RJ, Henderson AG, Alexis NE, Demaria G, Knight D, Thornton DJ, Sheehan JK (2009) Tracheobronchial air-liquid interface cell culture: a model for innate mucosal defense of the upper airways? Am J Physiol Lung Cell Mol Physiol 296(1):L92–L100PubMedCrossRefGoogle Scholar
  14. Kwilas S, Liesman RM, Zhang L, Walsh E, Pickles RJ, Peeples ME (2009) Respiratory syncytial virus grown in Vero cells contains a truncated attachment protein that alters its infectivity and dependence on glycosaminoglycans. J Virol 83(20):10710–10718PubMedCentralPubMedCrossRefGoogle Scholar
  15. Matrosovich MN, Matrosovich TY, Gray T, Roberts NA, Klenk HD (2004) Human and avian influenza viruses target different cell types in cultures of human airway epithelium. Proc Natl Acad Sci USA 101(13):4620–4624PubMedCrossRefGoogle Scholar
  16. Neilson KA, Yunis EJ (1990) Demonstration of respiratory syncytial virus in an autopsy series. Pediatr Pathol 10(4):491–502PubMedCrossRefGoogle Scholar
  17. Pickles RJ, McCarty D, Matsui H, Hart PJ, Randell SH, Boucher RC (1998) Limited entry of adenovirus vectors into well-differentiated airway epithelium is responsible for inefficient gene transfer. J Virol 72(7):6014–6023PubMedCentralPubMedGoogle Scholar
  18. Schaap-Nutt A, Scull MA, Schmidt AC, Murphy BR, Pickles RJ (2010) Growth restriction of an experimental live attenuated human parainfluenza virus type 2 vaccine in human ciliated airway epithelium in vitro parallels attenuation in African green monkeys. Vaccine 28(15):2788–2798Google Scholar
  19. Schaap-Nutt A, Liesman R, Bartlett EJ, Scull MA, Collins PL, Pickles RJ, Schmidt AC (2012) Human parainfluenza virus serotypes differ in their kinetics of replication and cytokine secretion in human tracheobronchial airway epithelium. Virology 433(2):320–328Google Scholar
  20. Scull MA, Gillim-Ross L, Santos C, Roberts KL, Bordonali E, Subbarao K, Barclay WS, Pickles RJ (2009) Avian Influenza virus glycoproteins restrict virus replication and spread through human airway epithelium at temperatures of the proximal airways. PLoS Pathog 5(5):e1000424PubMedCentralPubMedCrossRefGoogle Scholar
  21. Sims AC, Baric RS, Yount B, Burkett SE, Collins PL, Pickles RJ (2005) Severe acute respiratory syndrome coronavirus infection of human ciliated airway epithelia: role of ciliated cells in viral spread in the conducting airways of the lungs. J Virol 79(24):15511–15524PubMedCentralPubMedCrossRefGoogle Scholar
  22. Sims AC, Yount B, Burkett SE, Baric RS, Pickles RJ (2006) SARS CoV replication and pathogenesis in human airway epithelial cultures. Adv Exp Med Biol 581:535–538PubMedCrossRefGoogle Scholar
  23. Stonebraker JR, Wagner D, Lefensty RW, Burns K, Gendler SJ, Bergelson JM, Boucher RC, O’Neal WK, Pickles RJ (2004) Glycocalyx restricts adenoviral vector access to apical receptors expressed on respiratory epithelium in vitro and in vivo: role for tethered mucins as barriers to lumenal infection. J Virol 78(24):13755–13768PubMedCentralPubMedCrossRefGoogle Scholar
  24. Villenave R, Thavagnanam S, Sarlang S, Parker J, Douglas I, Skibinski G, Heaney LG, McKaigue JP, Coyle PV, Shields MD, Power UF (2012) In vitro modeling of respiratory syncytial virus infection of pediatric bronchial epithelium, the primary target of infection in vivo. Proc Natl Acad Sci USA 109(13):5040–5045Google Scholar
  25. Welliver TP, Reed JL, Welliver RC Sr (2008) Respiratory syncytial virus and influenza virus infections: observations from tissues of fatal infant cases. Pediatr Infect Dis J 27(10 Suppl):S92–S96PubMedCrossRefGoogle Scholar
  26. Wright PF, Ikizler MR, Gonzales RA, Carroll KN, Johnson JE, Werkhaven JA (2005) Growth of respiratory syncytial virus in primary epithelial cells from the human respiratory tract. J Virol 79(13):8651–8654PubMedCentralPubMedCrossRefGoogle Scholar
  27. Zabner J, Seiler M, Walters R, Kotin RM, Fulgeras W, Davidson BL, Chiorini JA (2000) Adeno-associated virus type 5 (AAV5) but not AAV2 binds to the apical surfaces of airway epithelia and facilitates gene transfer. J Virol 74(8):3852–3858PubMedCentralPubMedCrossRefGoogle Scholar
  28. Zhang L, Bukreyev A, Thompson CI, Watson B, Peeples ME, Collins PL, Pickles RJ (2005) Infection of ciliated cells by human parainfluenza virus type 3 in an in vitro model of human airway epithelium. J Virol 79(2):1113–1124PubMedCentralPubMedCrossRefGoogle Scholar
  29. Zhang L, Button B, Gabriel SE, Burkett S, Yan Y, Skiadopoulos MH, Dang YL, Vogel LN, McKay T, Mengos A, Boucher RC, Collins PL, Pickles RJ (2009) CFTR delivery to 25 % of surface epithelial cells restores normal rates of mucus transport to human cystic fibrosis airway epithelium. PLoS Biol 7(7):e1000155PubMedCentralPubMedCrossRefGoogle Scholar
  30. Zhang L, Collins PL, Lamb RA, Pickles RJ (2012) Comparison of differing cytopathic effects in human airway epithelium of parainfluenza virus 5 (W3A), parainfluenza virus type 3, and respiratory syncytial virus. Virology 421(1):67–77Google Scholar
  31. Zhang L, Peeples ME, Boucher RC, Collins PL, Pickles RJ (2002) Respiratory syncytial virus infection of human airway epithelial cells is polarized, specific to ciliated cells, and without obvious cytopathology. J Virol 76(11):5654–5666PubMedCentralPubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  1. 1.Department of Microbiology and ImmunologyUNC School of MedicineChapel HillUSA

Personalised recommendations