Novel intranasal GNRs-DNA vaccines against human papillomavirus infection

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.

Name dc:type.qualificationname

phd

Level dc:type.qualificationlevel

doctoral-pg

Grantor dc:publisher.institution

University of Strathclyde

Year dc:date.issued

2015

Author dc:creator

• Li, Wenqin

Advisors dc:contributor.advisor

• Yu, Jun
• Chen, Yu

DOI

10.48730/n72q-w388

Identifier

T14765

Author Identifier

201184117

OAI identifier oai:identifier

oai:strathclyde:9k41zd54n