Vesicular stomatitis virus Indiana strain is the prototype envelope glycoprotein for the genus Vesiculovirus. While the wild-type virus, VSVind, has developed into a potent and versatile oncolytic virotherapy and vaccine vector delivery platform, the G protein (VSVind.G) is ubiquitously used to pseudotype lentiviral vectors (LVs) for experimental and clinical applications. Recently, G proteins derived from other vesiculoviruses (VesG), for example, Cocal, Piry, and Chandipura viruses have been proposed as alternative LV envelopes with possible advantages compared to VSVind.G. However, vesiculovirus research has not developed extensively, and there is a gap in knowledge regarding the antigenic and immunogenic characteristic of VesG. In this work, I investigated two anti-VSVind.G monoclonal antibodies for their ability to cross-react with other VesG, identified the epitopes they recognise, and explored the mechanisms behind their neutralisation activity. Furthermore, these G proteins were characterised for their sensitivity to inactivation by fresh mammalian sera. Using some mix-and-match constructs, I identified that the hypervariable PH domain of VSVind.G confers sensitivity to otherwise serum resistant Cocal G. I further examined VesG regarding their immunogenicity, explored the humoral immune response triggered by systemic administration of LVs and investigated the inhibitory effects of the induced anti-G neutralising response on subsequent LV administrations. However, this could be alleviated using a heterogeneous panel of envelopes sequentially. Taken together, this work will broaden the use of VesG pseudotyped lentiviral vectors in clinical gene therapy by providing the proof-of-concept to circumvent anti-envelope immunity where repeated systemic vector administration is necessary and the opportunity to modify the VesG and improve G protein-containing advanced therapy medicinal products and vaccine vectors.
| Identifer | oai:union.ndltd.org:bl.uk/oai:ethos.bl.uk:763292 |
| Date | January 2018 |
| Creators | Munis, Altar Mert |
| Publisher | University College London (University of London) |
| Source Sets | Ethos UK |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
| Source | http://discovery.ucl.ac.uk/10060753/ |
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