Saudi Cultural Missions Theses & Dissertations
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Item Restricted Optimisation of Respiratory Syncytial Virus (RSV) whole genome sequencing of subtypes A and B(University of Nottingham, 2024-07) Alamoudi, Nouran Abubaker; King, Barnabas; Mcclure, PatrickRespiratory Syncytial Virus (RSV) is a major cause of lower respiratory tract infections, causing significant morbidity and mortality rates worldwide. RSV mainly affects infants, children under five years old, and older or immunocompromised adults. Establishing a cohesive method for the detection and identification of RSV could significantly improve monitoring and surveillance of RSV cases globally. The present study focused on optimising previously developed 400 base pair tiled-amplicon schemes for the whole genome sequencing of RSV subtypes A and B. We have developed novel ~1200 base pair primer schemes to increase sensitivity and produce a cost-efficient method for RSV genome sequencing. Samples were chosen between 2014 to 2024 and sequenced using Oxford Nanopore Technology’s PromethION device to acquire high- throughput genome data of both RSV subtypes. Our results revealed high potential for the optimised RSV A scheme, with all but two amplicons achieving genomic coverage depth of > 1000 reads per amplicon. However, the developed RSV B schemes generated insufficient results for most of RSV B samples. Moreover, the samples tested for RSV A displayed greater strain variability after phylogenetic analysis. In contrast, RSV B samples were of closely related strains and showed less genomic diversity. Acquiring high-throughput genomic data for RSV can hugely influence our understanding of the diversity of RSV, in addition to its evolutionary and seasonality patterns, which subsequently assists in the management of this virus on a global range. Furthermore, constituting a unified method for RSV sequencing largely impacts the implementation of successful surveillance systems for RSV infections.4 0Item Restricted Development of flexible hepatitis B virus core protein virus-like particles as a universal vaccine scaffold using a range of antigen-capturing systems(University of Leeds, 2024-07) Alzahrani, Jehad Abdulaziz; Stonehouse, Nicola; Stacey, MartinThe development of safe and flexible vaccines is an essential tool to fight emerging and re-emerging pathogens. Virus-like particles (VLPs) offer promising and adaptable options in vaccine development due to their safety and high immunogenicity. VLPs are non-infectious viral structures mimicking native viruses and can stimulate cellular and humoral immune responses. Additionally, VLPs can be modified as generic vaccine platforms to present various antigens. In this project, hepatitis B core antigen (HBcAg) VLPs were investigated as a presentation platform based on a fused dimer of two monomeric subunits. HBcAg VLPs were modified to incorporate antigen-capturing systems (ACSs) to create a new platform. The ACSs used here are small artificial non-antibody binding proteins that recognise and bind specific protein targets. The ACSs selected were an Affimer against small ubiquitin-like modifier (SUMO) tag, as well as the SpyTag/SpyCatcher systems. In model one, VLPs presented a single ACS: Affimer, SpyTag, or SpyCatcher at the N-terminus or the major immunodominant region (MIR) of HBcAg. In model two, dual antigen-capturing systems (dACSs) were used: Affimer was engineered into the MIR and SpyTag into the N-terminus. The dACS VLPs allow the decoration with dual antigens to enhance immunogenicity against a specific pathogen. These VLPs were successfully produced in a yeast expression system (Pichia pastoris, which is also known as Komagataella phaffii). In addition to the production of different VLPs, a fragment of glycoprotein G from respiratory syncytial virus (RSV-G) was also generated. The RSV-G protein was tagged with either SUMO tag or SpyCatcher to allow interaction with the corresponding VLP. RSV-G proteins were then successfully expressed in mammalian expression systems. The VLPs described above were decorated with a range of proteins, including glycoprotein 1 (GP1) of the Junín virus and glycoprotein G of the RSV-G. One of the VLPs was tested in an immunisation study in mice (with Junín virus GP1 as an antigen). The results showed superior responses for the decorated VLPs compared to the noncomplexed GP1. The results described in this thesis have demonstrated the development of a novel vaccine platform that could be employed with a number of different antigens in the future.24 0Item Restricted IL36 and IL37Cytokines, Mediators or Potential Modulators of Airway Infection and Inflammation?(Saudi Digital Library, 2022-10-01) Alshammari, Jamilah; Flannagan, BrianRespiratory tract infections caused by viruses or bacteria are among the most common human diseases worldwide. More specifically, respiratory syncytial virus (RSV) is a major cause of acute lower respiratory infection (ALRI), and this frequently results in hospitalisation in young children, particularly among those under the age of 5, and the elderly. Infection with RSV is also correlated with various airway respiratory diseases, such as bronchiolitis and asthma. In vivo testing suggests that levels of IL-36 alpha, IL-36 gamma, and IL-37 protein can be very high in nasopharyngeal aspirate (NPA) during RSV and rhinovirus (RV) infections. This work thus hypothesises that, during RSV and RV infections, the epithelial cells in the airways may express IL-36α, IL-36γ, and their respective receptors in a manner that plays a crucial pro-inflammatory role in RSV and RV infections. In addition, this work hypothesises the possibility of the further expression of IL-36Rα and IL-37 cytokines by airway epithelial cells (AECs) in response to RSV and RV infections. The main aim of this work, as described in this thesis, is thus to investigate whether RSV infection can trigger AECs to express IL-36 and IL-37 cytokines and their receptors.2 0