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Hinds, Joseph E (2025) Investigating lyssavirus glycoprotein interactions. PhD thesis, University of Nottingham.

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Abstract

The Lyssavirus genus contains the viruses responsible for the disease rabies, an acute progressive encephalomyelitis which is fatal in over 99% of human cases without treatment prior to the onset of symptoms. The genus is made up of three distinct phylogroups representing genetic distance and antigenic properties. The primary causative virus of rabies is the rabies virus which is involved in over 99% of human infections. Despite its highly pathogenic nature and the fact it causes over 59,000 deaths annually, particularly in developing nations, it is considered to be a neglected pathogen. While the human burden of lyssavirus infection is great, there are available treatments both pre and post infection. However, current treatments suffer from high costs, inconvenient and extensive treatment regimens, cold chain necessity and lack of inter-phylogroup neutralisation. These issues have generated interest in a novel vaccine.

For lyssaviruses, there is only one surface protein, the trimeric spike glycoprotein. This protein is involved in many important viral processes such as host receptor attachment and cell entry. It is because of this importance and the many remaining questions surrounding lyssaviruses that this study has focused on the glycoprotein and its interactions.

In order to study the glycoprotein in isolation a three plasmid pseudo-virus system was used. This system can be used to express a viral glycoprotein of interest on a vector such as a lentivirus in order to produce a non-replicative and safe pseudo-virus. This was used to infect immortalised cell lines such as BHK-21 to examine changes in outcome from both modifications of the glycoprotein and neutralisation using non-human primate sera or monoclonal antibodies. Additionally, this system was used to infect primary NK cells to determine their susceptibility to rabies virus infection.

Results of neutralisation assays performed by this study demonstrate that a novel medoid vaccine presented cross-phylogroup neutralising properties beyond any currently available vaccine, indicating a potential solution for divergent lyssavirus infection concerns. The vaccine was able to elicit significant neutralisation against MOKV and WCBV by week 5, and IKOV following a boost at week 50.

Glycosylation of the lyssavirus glycoprotein was also investigated and shown to be important in the infection capability and neutralisation susceptibility of the protein. The removal of fixed virus glycan Asn204 was found to significantly reduce infectivity and increase neutralisation susceptibility whereas removal of Asn37 increased infection though had no impact on neutralisation. The strongest change was in their combined removal. Asn319 removal also completely removed any infectivity. Common amino acids used in substitutions to knock out glycans aspartic acid (D), glutamine (Q) and alanine (A) were also examined for their impact and it was found that differing amino acids produced significantly different results. Finally, NK cell infection was found to not occur, however further work needs to be done to confirm this finding.

Research into rabies is both important and inadequate with many important questions remaining unanswered. The lyssavirus glycoprotein and its interactions are essential in understanding lyssaviruses and developing novel therapies for them. This study aimed to further elucidate some of the questions surrounding the glycoprotein. It also provides a foundation for more exciting discoveries and findings.

Item Type:	Thesis (University of Nottingham only) (PhD)
Supervisors:	Ball, Jonathan McClure, Patrick Urbanowicz, Richard
Keywords:	Rabies, Lyssavirus genus, Progressive encephalomyelitis
Subjects:	QS-QZ Preclinical sciences (NLM Classification) > QY Clinicial pathology
Faculties/Schools:	UK Campuses > Faculty of Medicine and Health Sciences > School of Life Sciences
Item ID:	82436
Depositing User:	Hinds, Joseph
Date Deposited:	31 Dec 2025 04:40
Last Modified:	31 Dec 2025 04:40
URI:	https://eprints.nottingham.ac.uk/id/eprint/82436

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