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- W2623355598 abstract "Background: Respiratory Syncytial Virus (RSV) is a common cause ofbronchiolitis during infancy; virtually all children are infected with this virus bythe age of 2 years. Although the majority have a self-limiting, mild infection, 1-3% have disease severe enough to warrant hospital admission. Thesechildren have an abundance of neutrophils (>80% of bronchoalveolar lavagecells) within their airways. There is however, a lack of detailed investigationinto their role in viral infection. Our group has previously shown RSVassociated with ex vivo neutrophils from infected infants. To explore thisobservation further, I have modelled and visualised the RSV-neutrophilinteraction, hypothesising that neutrophils play a role in the innate viralresponse in the airway.Aims: The aims of this thesis were to (i) establish an in vitro model of RSVneutrophilinteraction, (ii) determine whether RSV is taken up into theneutrophil, (iii) determine if RSV productively replicates inside the neutrophil,(iv) compare neutrophil-RSV interaction in adult and infant neutrophils and (v)investigate the response of the neutrophil to RSV.Methods: An in vitro model of RSV-neutrophil interaction was establishedusing highly purified adult and cord blood neutrophils. Neutrophils wereanalysed by quantitative RT-PCR, western blot, and by confocal microscopyusing indirect immunocytochemistry. In addition, ex vivo BAL neutrophils fromRSV infected infants were examined by confocal microscopy. Followingvalidation of this model, confirming the presence of virus within the neutrophil,a human gene microarray was performed.Results: Interaction of neutrophils with RSV was modelled in vitro anddemonstrated by PCR analysis and western blot. RSV N gene expressionrevealed maximal neutrophil uptake at 4 hours. A time course over 20 hoursshowed no increase in RSV expression, implying that active RSV replicationwas not occurring. Confocal microscopy revealed internalisation of RSV, withvirus distributed throughout the cytoplasm. Imaging of ex vivo neutrophilsextracted from bronchoalveolar lavage of RSV-infected infants showed asimilar distribution. RSV induced significant change in neutrophil geneexpression with over 1000 differentially expressed genes being identified.Pathway analysis revealed an RNA virus specific transcriptional response.Results for key molecules were subsequently validated by RT-PCR.Conclusions: Neutrophils do not allow RSV to replicate within them. Incontrast, following interaction with RSV, they alter their gene expressionprofiles through the modulation of interferon responses and other pathways,thereby inhibiting viral replication. Evidence presented in this thesis supportsa novel role for neutrophils as innate cells at the frontline of antiviral immunityin RSV disease." @default.
- W2623355598 created "2017-06-15" @default.
- W2623355598 creator A5018025847 @default.
- W2623355598 date "2015-01-01" @default.
- W2623355598 modified "2023-09-23" @default.
- W2623355598 title "In vitro characterisation of the neutrophil in respiratory syncytial virus bronchiolitis" @default.
- W2623355598 doi "https://doi.org/10.17638/02000681" @default.
- W2623355598 hasPublicationYear "2015" @default.
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