The collectins surfactant protein A (SP-A) and SP-D are found in the lining fluid of the lung and form an important component of innate pulmonary defenses; they consist of a N-terminal region, a collagenous region, a neck region and a carbohydrate recognition domain (CRD). SP-A and SP-D bind to and have antiviral properties in vivo against the enveloped viruses respiratory syncytial virus (RSV) and influenza A virus (IAV). A recombinant fragment of SP-D (rfhSP-D) has also been developed and is composed of the neck, CRD and a short collagenous region of SP-D. The rfhSP-D retains the ability to bind to RSV and IAV. Human rhinovirus (HRV) is a non-enveloped virus and is the predominant cause of the common cold. HRV can also cause exacerbations in patients with chronic airway disease. Based on the known antiviral effects of SP-A and SP-D, and their localisation in the upper and lower airways, the present thesis sought to investigate the research question 'Can SP-A, SP-D and rfhSP-D bind to and inhibit infectivity of HRV?' Methods to purify and detect HRV were established and used in experiments with SP-A and SP-D purified from bronchoalveolar lavage and rfhSP-D expressed in Escherichia coli. A novel interaction between SP-D and rfhSP-D with HRV was identified using co-irnmunoprecipitation and surface plasmon resonance; no such interaction was detected with SP-A. Binding was calcium dependent and inhibited by the presence of sugars, indicating that binding is via the CRD. Monolayers of HeLa cells, 16HBE cells or undifferentiated primary nasal epithelial cells were infected with HRV coincubated with collectins and the infection level was assessed using flow cytometry. The presence of collectins did not significantly alter the infection level. This is the first report of a component of the innate immune system binding to the surface of HRV.
| Identifer | oai:union.ndltd.org:bl.uk/oai:ethos.bl.uk:588802 |
| Date | January 2012 |
| Creators | Pugh, Jacqueline |
| Contributors | Clark, Howard ; Madsen, Jens |
| Publisher | University of Southampton |
| Source Sets | Ethos UK |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
| Source | https://eprints.soton.ac.uk/360439/ |
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