Macrophages are required for dendritic cell uptake of respiratory syncytial virus from an infected epithelium

PLoS One. 2014 Mar 20;9(3):e91855. doi: 10.1371/journal.pone.0091855. eCollection 2014.

Abstract

We have previously shown that the respiratory syncytial virus [RSV] can productively infect monocyte derived dendritic cells [MoDC] and remain dormant within the same cells for prolonged periods. It is therefore possible that infected dendritic cells act as a reservoir within the airways of individuals between annual epidemics. In the present study we explored the possibility that sub-epithelial DCs can be infected with RSV from differentiated bronchial epithelium and that in turn RSV from DCs can infect the epithelium. A dual co-culture model was established in which a differentiated primary airway epithelium on an Air Liquid Interface (ALI) was cultured on a transwell insert and MoDCs were subsequently added to the basolateral membrane of the insert. Further experiments were undertaken using a triple co-culture model in which in which macrophages were added to the apical surface of the differentiated epithelium. A modified RSV [rr-RSV] expressing a red fluorescent protein marker of replication was used to infect either the MoDCs or the differentiated epithelium and infection of the reciprocal cell type was assessed using confocal microscopy. Our data shows that primary epithelium became infected when rr-RSV infected MoDCs were introduced onto the basal surface of the transwell insert. MoDCs located beneath the epithelium did not become infected with virus from infected epithelial cells in the dual co-culture model. However when macrophages were present on the apical surface of the primary epithelium infection of the basal MoDCs occurred. Our data suggests that RSV infected dendritic cells readily transmit infection to epithelial cells even when they are located beneath the basal layer. However macrophages appear to be necessary for the transmission of infection from epithelial cells to basal dendritic cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Cell Differentiation
  • Coculture Techniques
  • Dendritic Cells / pathology
  • Dendritic Cells / virology*
  • Epithelial Cells / pathology
  • Epithelial Cells / virology
  • Epithelium / pathology
  • Epithelium / virology*
  • Fluorescence
  • Humans
  • Macrophages / pathology
  • Macrophages / virology*
  • Monocytes / pathology
  • Reproducibility of Results
  • Respiratory Syncytial Virus Infections / pathology
  • Respiratory Syncytial Virus Infections / virology*
  • Respiratory Syncytial Viruses / physiology*
  • Virus Replication

Grants and funding

The study was funded by the Sheffield Children’s Hospital Research Committee. The funders had no role in study design, data collection, analysis, decision to publish or preparation of the manuscript.