Use this URL to cite or link to this record in EThOS: http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.695259
Title: Exploitation of well-differentiated primary paediatric airway epithelial cell cultures (WD-PAECs) to study RSV/human host interactions
Author: Broadbent , Lindsay Jane
ISNI:       0000 0004 5994 9181
Awarding Body: Queen's University Belfast
Current Institution: Queen's University Belfast
Date of Award: 2016
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Abstract:
Respiratory syncytial virus (RSV) is the primary cause of severe lower respiratory tract infections in children. We exploited well-differentiated primary paediatric airway epithelial cells (WD-PAECs) to investigate innate immune responses to RSV. We demonstrated that RSV-conditioned medium (CMRSV) treated WD-PAECs were highly resistant to subsequent RSV or Sendai virus infection. IL-29 significantly contributes to the antiviral activity of CMRSV. Apical pre-treatment, but not basolateral pre-treatment, of WD-PBECs with IL-29 decreased RSV growth kinetics. We found that IL28RA, a component of the type III IFN receptor, was located exclusively on the apical surface of WD-PBECs. We demonstrated that type IIllFNs modulate the innate immune responses to RSV infection. Pre-treatment of WD-PBECs with a TLR4 signalling inhibitor resulted in massive reductions in RSV growth kinetics, and IL-29 and pro-inflammatory cytokine secretion. To elucidate the role of TLR4 in RSV infection we used inhibitors of TLR4 pathway intermediates. Our results showed it is likely that RSV-induced IL-29 is mediated, in part, by TLR4 signalling through NF-KB and/or p38 MAPK. We identified ten genes that were significantly upregulated in RSV-infected WD-PAEC cultures. Furthermore we identified nine genes that are differentially expressed between two patient cohorts: history of mild or severe RSV disease. The proteins encoded by isg15, rsad2 and tnfsf13b, were assessed for direct antiviral properties. However, they failed to reduce viral replication. The gene ptn was found to be differentially expressed in WD-PNECs from the two cohorts irrespective of RSV infection. Pre-treatment of WD-PBECs at 40C with PTN significantly reduced RSV replication. In summary, we demonstrated that RSV induced antiviral responses are mediated, in part, by IL-29. TLR4 signalling plays a significant role in RSV replication and induction of IL-29 and pro-inflammatory chemokines. We identified three potential anti-RSV molecules: IL-29, CLI-095 and PTN, which may be beneficial in the battle against RSV.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID: uk.bl.ethos.695259  DOI: Not available
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