Characterization and Development of a Novel Cytomegalovirus for Evaluation as an HIV Vaccine Viral Vector in a Non-Human Primate Model

Within the field of HIV vaccine design, it has been recognized that there is a real need to diversify the pool of vectors undergoing testing and to utilize vectors that can induce durable immunity. Herpesviruses, such as cytomegalovirus (CMV), show much promise as viral vectors, as they establish lifelong latency with periodic reactivation, enabling persistent expression of vaccine antigens. We propose that using CMV as a lifelong reactivating HIV viral vector will generate a robust and persistent effector memory T cell response at the mucosa and thus, may impair HIV replication at its earliest stage. In an effort to evaluate CMV-based HIV/SIV vectors in a non-human primate (NHP) model, I isolated CMV from cynomolgus macaques (CyCMV), and performed preliminary phenotypic and genomic classification of the novel virus. To characterize the CyCMV genome, facilitate targeted recombination for vaccine construction, and inform possible attenuation strategies, I sequenced and annotated the complete viral genome using next-generation sequencing. Furthermore, I compared and contrasted the structural and functional genes of CyCMV to human and rhesus macaque CMV with respect to genes involved in pathogenesis, immune evasion, species-specificity, and more. In order to maximize the ability to manipulate the virus for vaccine purposes, I cloned CyCMV as a Bacterial Artificial Chromosome (BAC). The cloning of CyCMV-BAC enabled the virus to be transformed into a bacterial system for downstream insertion of vaccine antigens, as well as future viral manipulations, including attenuations. This newly characterized CMV provides a novel model in which to study HIV/SIV CMV-based vaccines in hopes of informing forthcoming human clinical trials. Additionally, CyCMV-BAC will provide an invaluable resource for CMV biologists allowing for virus manipulation studies, functional characterization of CMV genes, as well as HCMV pathogenesis and vaccine development research.

Department dc:contributor.department

Immunology

Year dc:date.issued

2014

Author dc:creator

• Marsh, Angie

Advisor dc:contributor.advisor

• MacDonald, Kelly

• HIV
• Vaccine Viral Vector
• Cytomegalovirus
• Non-Human Primate
• Herpesvirus
• Cynomolgus Macaque

Statement dc:rights

• Attribution 2.5 Canada

Licence dc:rights.uri

• http://creativecommons.org/licenses/by/2.5/ca/

Language dc:language.iso

en_ca

Handle dc:identifier.uri

http://hdl.handle.net/1807/69060

OAI identifier oai:identifier

oai:utoronto.scholaris.ca:1807/69060