The world-wide dissemination of the human immunodeficiency virus type 1 (HIV-1) has led to a wide number of approaches aimed at controlling and understanding viral replication. Although antiviral drugs have been important in the treatment of HIV-1-infected individuals, toxicity and resistance have mandated new efforts and strategies. / We have tried to develop a trans-dominant gene therapy approach, by showing that mutated Gag-Pol polyproteins can be incorporated into viral particles. In our studies, Gag-Pol proteins that were mutated in protease (PR) and combinations of mutations in PR and reverse transcriptase (RT) inhibited the production of fully processed and infectious viral particles when coexpressed with the infectious HIV-1 molecular clone, pBH10. Viral particles produced after cotransfection of COS-7 cells with both pBH10 and Gag-Pol, mutated in PR alone or in both RT and PR, showed abnormal processing and lower infectivity, suggesting that a trans-dominant negative effect had occurred. These effects were not seen when Gag-Pol with a deleted integrase ($ Delta$IN) and the same RT and PR mutations were co-expressed with pBH10. Complementation experiments in which pBH10 mutated in PR was co-expressed with wild-type Gag-Pol showed that the latter could be incorporated into viral particles. Finally, COS-7 cells stably transfected with Gag-Pol mutated in PR or in both PR and RT, and subsequently transfected with pBH10, showed diminished production of viral particles.
| Identifer | oai:union.ndltd.org:LACETR/oai:collectionscanada.gc.ca:QMM.27893 |
| Date | January 1997 |
| Creators | Morin, Nicolas. |
| Contributors | Wainber, Mark A. (advisor) |
| Publisher | McGill University |
| Source Sets | Library and Archives Canada ETDs Repository / Centre d'archives des thèses électroniques de Bibliothèque et Archives Canada |
| Language | English |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
| Format | application/pdf |
| Coverage | Master of Science (Department of Microbiology and Immunology.) |
| Rights | All items in eScholarship@McGill are protected by copyright with all rights reserved unless otherwise indicated. |
| Relation | alephsysno: 001601370, proquestno: MQ37152, Theses scanned by UMI/ProQuest. |
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