Thesis

Novel intranasal GNRs-DNA vaccines against human papillomavirus infection

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Creator
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Awarding institution
  • University of Strathclyde
Date of award
  • 2015
Thesis identifier
  • T14765
Person Identifier (Local)
  • 201184117
Qualification Level
Qualification Name
Department, School or Faculty
Abstract
  • Currently licensed HPV vaccines, Cervarix and Gardasil, have been successfulin preventing HPV infection and high-graded cervical intraepithelial neoplasia (CIN2,3) in school girls in the UK and other developed countries. However, their high costshave hindered their wider use in preventing HPV infection and CIN 2/3 associatedwith HPV type -16 and -18. This project aims to produce cheap and effective GNRsDNA vaccines. The vaccines were composed of two parts, gold nanorods and circularplasmid DNA that carries codon-optimized HPV type 16 L1 gene for expression ofthe viral capsid-like particles in host cells. GNRs were functionalized with either PEIor PDDAC cationic polymers for high affinity to negatively charged DNAs. TheDNA is constructed in the backbone of a mammalian expression vector, plasmidpcDNA3 YFP of which YFP offered a fluorescent indicator. The HPV type -16 L1was either expressed alone or together with E. coli heat labile toxin B subunit, a wellknown nasal adjuvant that enhances immunogenicity. In total, four GNRs-DNAwere produced: PEI-GNRs-pcDNA3 YFP L1 (PE-G-L), PEI-GNRs-pcDNA3 YFPL1-ELT (PE-G-EL), PDDAC-GNRs-pcDNA3 YFP L1 (PD-G-L), and PDDACGNRs-pcDNA3 YFP L1-ELT (PD-G-EL).The GNRs-DNA internalization led to expression of VLP in HEK293, HeLaand the murine bone marrow-derived macrophages cell. Intranasal immunization ofBALB/c mice with GNRs-DNA resulted in YFP expression in nasal and lung tissuesin one day. Three doses (3 × 480ng plasmid DNA per mouse) elicited neutralizingantibodies in the serum and antibody secreting cells in spleen lymphocytes specificto HPV16 L1 protein. PDG-YL appeared to be most potent and comparable topurified VLP protein in eliciting Nabs and ASCs.My work has demonstrated the feasibility of using GNRs-DNA as vaccineplatforms. GNRs-DNA is cheap to produce, easy to administer and stable at roomtemperature. These characteristics make GNRs-DNA attractive candidates for futureHPV vaccine development. The optimised concentration ratio of GNRs and DNA, Tcell immune-response, IgA secretion require further assessment before the clinicaltrial.
Advisor / supervisor
  • Yu, Jun
  • Chen, Yu
Resource Type
Note
  • Previously held under moratorium from 21st November 2017 until 21st November 2022.
DOI
Date Created
  • 2015
Former identifier
  • 9912573192602996

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