Objectives: The aim of this study was to investigate the development and progression of phenotypic resistance to the HIV-1-reverse transcriptase (RT) inhibitor lamivudine, and genotypic variations of HIV-1-RT occurring under lamivudine treatment in HIV-1-infected human primary monocytes-macrophages (M/M). Methods: Cellular passages in the presence of lamivudine were performed every 2 weeks by transferring supernatants of infected M/M to fresh M/M. A fitness assay using wild-type virus and a lamivudine-resistant HIV-1 virus (harbouring the M184V RT mutation) was performed in peripheral blood mononuclear cells. Culture supernatants were tested for p24 antigen production and RT activity. The M184V RT mutant virus was obtained by site-directed mutagenesis on a CCR5-using HIV-1 backbone. Results: The mutagenized M184V RT virus showed full resistance to lamivudine in M/M. However, no detectable phenotypic and genotypic resistance (neither virus breakthrough, nor RT resistance-related mutations) developed in M/M infected by HIV-1 and cultured for up to seven passages in vitro (i.e. 105 days). This inefficiency of M/M to develop M184V RT mutated virus is tightly related to the low 2'-deoxynucleotide (dNTP) pool in such cells, which in turn decreases the kinetics of HIV-1-RT. Despite this, the M184V RT mutant virus replicates in M/M, although with a 30% decreased efficiency compared with the wild-type. Conclusions: Our results show that the chances of development of resistance are far lower in M/M than in lymphocytes. This underlines the importance and the peculiar role of M/M as reservoirs of either wild-type or resistant strains in human organs.

Aquaro, S., Svicher, V., CECCHERINI SILBERSTEIN, F., Cenci, A., Marcuccilli, F., Giannella, S., et al. (2005). Limited development and progression of resistance of HIV-1 to the nucleoside analogue reverse transcriptase inhibitor lamivudine in human primary macrophages. JOURNAL OF ANTIMICROBIAL CHEMOTHERAPY, 55(6), 872-878 [10.1093/jac/dki104].

Limited development and progression of resistance of HIV-1 to the nucleoside analogue reverse transcriptase inhibitor lamivudine in human primary macrophages

Svicher, V;CECCHERINI SILBERSTEIN, FRANCESCA;PERNO, CARLO FEDERICO
2005-01-01

Abstract

Objectives: The aim of this study was to investigate the development and progression of phenotypic resistance to the HIV-1-reverse transcriptase (RT) inhibitor lamivudine, and genotypic variations of HIV-1-RT occurring under lamivudine treatment in HIV-1-infected human primary monocytes-macrophages (M/M). Methods: Cellular passages in the presence of lamivudine were performed every 2 weeks by transferring supernatants of infected M/M to fresh M/M. A fitness assay using wild-type virus and a lamivudine-resistant HIV-1 virus (harbouring the M184V RT mutation) was performed in peripheral blood mononuclear cells. Culture supernatants were tested for p24 antigen production and RT activity. The M184V RT mutant virus was obtained by site-directed mutagenesis on a CCR5-using HIV-1 backbone. Results: The mutagenized M184V RT virus showed full resistance to lamivudine in M/M. However, no detectable phenotypic and genotypic resistance (neither virus breakthrough, nor RT resistance-related mutations) developed in M/M infected by HIV-1 and cultured for up to seven passages in vitro (i.e. 105 days). This inefficiency of M/M to develop M184V RT mutated virus is tightly related to the low 2'-deoxynucleotide (dNTP) pool in such cells, which in turn decreases the kinetics of HIV-1-RT. Despite this, the M184V RT mutant virus replicates in M/M, although with a 30% decreased efficiency compared with the wild-type. Conclusions: Our results show that the chances of development of resistance are far lower in M/M than in lymphocytes. This underlines the importance and the peculiar role of M/M as reservoirs of either wild-type or resistant strains in human organs.
2005
Pubblicato
Rilevanza internazionale
Articolo
Sì, ma tipo non specificato
Settore MED/07 - MICROBIOLOGIA E MICROBIOLOGIA CLINICA
English
Con Impact Factor ISI
deoxyribonucleotide; lamivudine; nucleoside analog; RNA directed DNA polymerase; RNA directed DNA polymerase inhibitor; antibiotic resistance; article; controlled study; enzyme activity; genetic resistance; genetic variability; genotype; human; human cell; Human immunodeficiency virus 1; Human immunodeficiency virus infection; in vitro study; macrophage; nonhuman; peripheral blood mononuclear cell; phenotype; site directed mutagenesis; supernatant; virus mutant; virus mutation; virus replication; wild type; Anti-HIV Agents; Cells, Cultured; Drug Resistance, Viral; HIV-1; HIV-1 Reverse Transcriptase; Humans; Lamivudine; Macrophages; Mutation; Reverse Transcriptase Inhibitors
Aquaro, S., Svicher, V., CECCHERINI SILBERSTEIN, F., Cenci, A., Marcuccilli, F., Giannella, S., et al. (2005). Limited development and progression of resistance of HIV-1 to the nucleoside analogue reverse transcriptase inhibitor lamivudine in human primary macrophages. JOURNAL OF ANTIMICROBIAL CHEMOTHERAPY, 55(6), 872-878 [10.1093/jac/dki104].
Aquaro, S; Svicher, V; CECCHERINI SILBERSTEIN, F; Cenci, A; Marcuccilli, F; Giannella, S; Marcon, L; Calio, R; Balzarini, J; Perno, Cf
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2108/42095
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