Thesis

Novel intranasal GNRs-DNA vaccines against human papillomavirus infection

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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 successful in preventing HPV infection and high-graded cervical intraepithelial neoplasia (CIN2, 3) in school girls in the UK and other developed countries. However, their high costs have hindered their wider use in preventing HPV infection and CIN 2/3 associated with 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 circular plasmid DNA that carries codon-optimized HPV type 16 L1 gene for expression of the viral capsid-like particles in host cells. GNRs were functionalized with either PEI or PDDAC cationic polymers for high affinity to negatively charged DNAs. The DNA is constructed in the backbone of a mammalian expression vector, plasmid pcDNA3 YFP of which YFP offered a fluorescent indicator. The HPV type -16 L1 was either expressed alone or together with E. coli heat labile toxin B subunit, a wellknown nasal adjuvant that enhances immunogenicity. In total, four GNRs-DNA were produced: PEI-GNRs-pcDNA3 YFP L1 (PE-G-L), PEI-GNRs-pcDNA3 YFP L1-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, HeLa and the murine bone marrow-derived macrophages cell. Intranasal immunization of BALB/c mice with GNRs-DNA resulted in YFP expression in nasal and lung tissues in one day. Three doses (3 × 480ng plasmid DNA per mouse) elicited neutralizing antibodies in the serum and antibody secreting cells in spleen lymphocytes specific to HPV16 L1 protein. PDG-YL appeared to be most potent and comparable to purified VLP protein in eliciting Nabs and ASCs. My work has demonstrated the feasibility of using GNRs-DNA as vaccine platforms. GNRs-DNA is cheap to produce, easy to administer and stable at room temperature. These characteristics make GNRs-DNA attractive candidates for future HPV vaccine development. The optimised concentration ratio of GNRs and DNA, Tcell immune-response, IgA secretion require further assessment before the clinical trial.
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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