Toon, Kamilla;
(2022)
Production and evaluation of pseudotyped viruses for Hepacivirus and Flavivirus studies.
Doctoral thesis (Ph.D), UCL (University College London).
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Abstract
Flaviviridae is a diverse family of RNA viruses that includes many important human pathogens such as dengue virus, West Nile virus, Japanese encephalitis virus (JEV), and hepatitis C virus (HCV). Handling many of these pathogens requires high containment level biosafety laboratories and some cannot replicate in cell culture. The use of replication deficient pseudotyped viruses (PV) circumvents these issues. This thesis shows the development of two Flaviviridae PVs, GB Virus B (GBV-B) and JEV, for serological assay development and virus-host interaction studies. HCV is a major global concern with over 70 million people chronically infected. Until the recent discovery of related animal hepaciviruses, GBV-B was the only known homolog of HCV and is used in new world monkeys as an animal model. However, there is still little known about the viral-host interactions. This thesis first describes the production and optimization of GBV-B PV to develop a neutralization assay for screening sera from experimentally infected tamarins. Viral entry mechanisms were investigated using the GBV-B PV. Cells without Claudin 1 (CLDN1), a known HCV receptor, had no detectible GBV-B infection. CLDN1 was subsequently confirmed to be an entry factor when restoration of expression conferred susceptibility to GBV-B PV. Chimeric claudin proteins consisting of the permissive CLDN1 and non-permissive CLDN9 were created to investigate the region of importance for GBV-B and CLDN1 interaction. The important region was identified as extracellular loop 2, or downstream, which is different from the dependence of CLDN1-restricted HCV strains on regions in extracellular loop 1. Research with JEV requires high containment level biosafety labs which are often unavailable in resource limited areas. Described here is the production of JEV PV using two alternative pseudotyping methods, in vitro assembly of subviral particles with envelope-less lentiviral particles and dengue virus core, after failed attempts at using traditional systems.
| Type: | Thesis (Doctoral) |
|---|---|
| Qualification: | Ph.D |
| Title: | Production and evaluation of pseudotyped viruses for Hepacivirus and Flavivirus studies |
| Open access status: | An open access version is available from UCL Discovery |
| Language: | English |
| Additional information: | Copyright © The Author 2022. Original content in this thesis is licensed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International (CC BY-NC 4.0) Licence (https://creativecommons.org/licenses/by-nc/4.0/). Any third-party copyright material present remains the property of its respective owner(s) and is licensed under its existing terms. Access may initially be restricted at the author’s request. |
| UCL classification: | UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences > Div of Infection and Immunity UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences UCL |
| URI: | https://discovery.ucl.ac.uk/id/eprint/10153841 |
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