title: Primary airway epithelial cultures from children are highly permissive to respiratory syncytial virus infection.
abstract: BACKGROUND
  Respiratory syncytial virus (RSV) infection of airway epithelial cells (AECs) is an important initial event in RSV bronchiolitis. AEC immunological responses are thought to be critical in driving the subsequent inflammation in the airway. This study examined viral replication, cytotoxicity and cytokine production in cultures of primary AECs from children compared with responses to RSV infection in an immortalised epithelial cell line and to those from infants with RSV bronchiolitis.
  
  
  METHODS
  RSV replication, proinflammatory cytokine responses and cytotoxicity in RSV-infected primary AEC cultures derived from bronchial brushings from the lungs of children were compared with those seen in BEAS-2B cultures, as well as AECs and bronchoalveolar lavage fluid collected from children with and without RSV bronchiolitis.
  
  
  RESULTS
  Viral replication, cytotoxicity and inflammatory cytokine production were greater in primary AEC cultures than in BEAS-2B cells. Different response patterns were observed, with RSV infection of primary AEC cultures causing distinct peaks of viral replication and matched cytotoxic responses. Some primary AEC culture immunological responses, such as interleukin 8, were similar in magnitude to those seen in clinical samples from the lungs of children with RSV bronchiolitis. Although variable amounts of RSV were detected by PCR in freshly isolated primary AECs, RSV was not detected by immunocytochemistry.
  
  
  CONCLUSIONS
  This is one of the first studies to examine comprehensively the responses to RSV infection in primary AEC cultures from children and shows marked differences from those of a commercially available immortalised human cell line but reassuring similarities to results found in vivo. This suggests that future work investigating responses of AECs to RSV infection should use primary AEC cultures.
authors.affiliation: Institute of Translational Medicine (Child Health), University of Liverpool, Alder Hey Children's Hospital, Eaton Road, Liverpool L12 2AP, UK. a.fonceca@liv.ac.uk
authors.forename: A M
authors.initials: AM
authors.lastname: Fonceca
authors.forename: B F
authors.initials: BF
authors.lastname: Flanagan
authors.forename: R
authors.initials: R
authors.lastname: Trinick
authors.forename: R L
authors.initials: RL
authors.lastname: Smyth
authors.forename: P S
authors.initials: PS
authors.lastname: McNamara
mesh_terms: D000914:Antibodies, Viral / Q000032:analysis; D001980:Bronchi / Q000473:pathology* / Q000821:virology; D001992:Bronchoalveolar Lavage Fluid / Q000821:virology; D002460:Cell Line; D002648:Child; D002675:Child, Preschool; D016207:Cytokines / Q000096:biosynthesis; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D007223:Infant; D007231:Infant, Newborn; D008297:Male; D011379:Prognosis; D020545:Respiratory Mucosa / Q000378:metabolism / Q000473:pathology* / Q000821:virology; D018357:Respiratory Syncytial Virus Infections / Q000276:immunology / Q000473:pathology* / Q000821:virology; D012136:Respiratory Syncytial Viruses / Q000302:isolation & purification / Q000502:physiology*; D014779:Virus Replication
chemical_list: D000914:Antibodies, Viral; D016207:Cytokines