Abstract
Feline coronavirus (FCoV) leads to the fatal disease known as feline infectious peritonitis in a small proportion of infected cats. Research into FCoV has so far been hindered by our inability to culture the most common serotype, Type 1, in vitro.This project aimed to establish a reverse genetic system for Type 1 FCoV. Such a system would enable a deeper understanding of the role of viral mutations in the pathogenesis of disease and could serve as a platform for rational vaccine design. To this end, cDNA infectious clone and replicon constructs based on Type 1 FCoV were developed and transcribed. During this project, infectious virus or replicon-expressing cells were not recovered following transfection of the constructs into mammalian cells, but a foundation was laid for this to be achieved in future.
Recovery of recombinant Type 1 FCoV would necessitate a cell line capable of supporting this serotype’s growth in vitro, so an aim of this project was to identify a cell entry receptor (CER) for Type 1 FCoV in order to develop a cell line permissive to infection with the virus. To this end, ‘bait proteins’ bearing the spike proteins of Type 1 and 2 FCoV were produced, and Type 2 bait protein was able to recognise its CER. Feline intestinal organoid cultures were established, and an interaction identified between Type 1 bait protein and heat shock 70 kDa protein 1a (HSPA1A) suggested that HSPA1A is a receptor for Type 1 FCoV. Finally, a feline IFN-gamma ELISpot assay was used to measure the cellular immune response to peptides representing a Type 1 FCoV epitope with and without a substitution. No significant difference in the ability of the peptide variants to stimulate a response was identified, but the assay could be used in the future to guide rational vaccine design.
| Date of Award | 23 Jan 2020 |
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| Original language | English |
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| Supervisor | Andrew D Davidson (Supervisor) & Severine Tasker (Supervisor) |