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Touabi, Lila (2024) Rhinovirus neutralisation and the role of TRIM21. Doctoral thesis, London Metropolitan University.
Human Rhinoviruses (RV) are small viruses that cause the majority of common colds and have a huge impact on human health. Currently there is no effective vaccine against these viruses which is largely due to the antigenic diversity amongst approximately 160 stereotypes. However, studies have shown that antibodies are able to offer some cross-serotype protection, despite significant sequence variations in the surface-exposed areas of capsid proteins VP1, VP2 and VP3. Antibodies are known to neutralise rhinovirus through four immunogenic epitopes mapping to these most variable exposed regions.
Conventional antibodies can neutralise viruses in a variety of ways, including by aggregating viral particles to reduce interactions with receptors and prevent attachment to host cells. Antibodies can also bind to viruses to prevent them from attaching to host cells by steric hindrance. Antibodies can prevent the release of RNA into cells by stabilising the capsid or cause conformational changes of the capsid, which prevents virions from attaching to host cell receptors and prematurely starts the release of RNA. Another method of protection against non-enveloped viruses such as rhinoviruses is neutralisation through the antiviral protein TRIM2 l. When viruses enter the cytosol, TRIM21 can bind to the FC region of the antibodies attached to viruses and trigger the proteasome to degrade the virions by releasing free K63 polyubiquitin chains.
The aim of this research was to generate neutralising polyclonal and monoclonal antibodies to RV and identify the role of TRIM21 in RV neutralisation by determining if monoclonal antibodies generated require TRIM21 for intracellular RV neutralisation and by analysing the expression of TRIM21 mRNA in response to RV infection. Additionally, this study intended to investigate the dominant VP2-P6 epitope that may induce protective cross-serotype antibody responses. We have generated mouse polyclonal antibodies through different mouse immunisations and human monoclonal antibodies by phage display technology panning for binding to dominant VP2-P6 epitope. However, these antibodies are serotype specific and non-neutralising. These antibodies could not be applied to investigate the role of TRIM21 in RV intracellular neutralisation as they do not neutralise RV, but this study demonstrated that TRIM21 is a virus inducible gene as TRIM21 was upregulated upon RV infection at low and high MOI in HeLa and BEAS-2B cells. However, further studies are required to investigate the role of TRIM21 in RV intracellular neutralisation in the presence of neutralising monoclonal antibodies.
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