610. The Effect of In Vitro Antiviral Therapy on Lentiviral Transduction
Year: 2006
Type: article
Abstract: With the advent of lentivirus vector (LV)-based gene transfer research, it is important to understand the potential effect of commonly used anti-HIV agents on lentivirus transduction efficiency. One would expect that a nucleoside reverse transcriptase inhibitor (NRTI) would inhibit LV-transduction, and one commonly used NRTI agent, tenofovir (PMPA) has a prolonged intracellular half-life. To explore the influence of NRTI drugs and protease inhibitors on lentiviral transduction, cord blood CD34 were treated with either PMPA, zidovudine (AZT), or indinavir before transduction with a recombinant lentivirus vector expressing eGFP. Cells were treated with the antiviral agents in vitro for 24hr and then the agents were removed from the media and lentivirus transductions were done at day 1, 3, and 9 post-treatment. The persistence of transduction inhibition after drug removal was determined, and these results were compared with the TCID50 of these agents against HIV strain IIIB. For the clinical HIV-1 challenge, virus was added at the time of drug removal. The TCID50 for PMPA, AZT, and indinavir vs HIV-1 by p24 ELISA were (1.4–3.2) μM, 27 nM and (5.2–58) nM respectively. The effects of the agents on transduction efficiency are shown in the table below. Both PMPA and AZT reduced the level of post-transduction gene expression 24-hr after drug removal by 69% and 86%, respectively, and this effect persisted for three days (28% and 86% inhibition, respectively). By day 9 post-treatment, the gene transfer efficiency had returned to 92% and 81%, respectively, of the untreated control. To determine if the decrease in gene transfer was due to drug toxicity, the cells were placed in methylcellulose colony forming assay. PMPA did not affect the number of CFUs but AZT did have an inhibitory effect (see ) which lasted until day 5. Indinavir had only a small effect on transduction at day 1 post-treatment. To further explore the effect of these antivirals on transduction efficiency, PMPA or indinavir were added at 2, 24, 36 72, and 96 hrs AFTER transduction. There was a 40% reduction by PMPA treatment at 2 hrs but no further reductions were observed. Tabled 1Percentage of Inhibition of lentiviral transduction by anti-HIV-1 agentsDays post RxUntreatedPMPA 5 uMAZT 100 uMIndinavir 1 uMDay 106 98 62 7Day 302 88 30Day 502 77 90Day 9081 90 Open table in a new tab Tabled 1Effect of AZR and PMPA on hematopoietic progenitor colonies in methylcelluloseDays post RxmockuntreatedPMPA 5 uMAZT 100 uMDay 11091021131 4Day 31371247 23 9Day 59 21658 4108 Open table in a new tab In conclusion, PMPA and AZT exposure pre-treatment can significantly inhibit tbransduction efficiency by lentivirus vectors, and this effect lasts for up to one week. There is little effect after 2 hours post-transduction. In gene transfer research studies involving HIV/AIDS patients, it is warranted to stop PMPA, AZT, and other NRTI therapies before blood cells are collected for the research transductions.
Source: Molecular Therapy
Institution: Beckman Research Institute
Language: English
Subfield: Genetics
Domain: Life Sciences
Open Access status: hybrid
Estimated APC paid: $4,500
OpenAlex ID: W1983307643