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T20-insensitive HIV-1 from naive patients exhibits high viral fitness in a novel dual-color competition assay on primary cells.
- Source :
-
Virology [Virology] 2005 Mar 15; Vol. 333 (2), pp. 251-62. - Publication Year :
- 2005
-
Abstract
- The relationship between sensitivity to antiviral drugs and viral fitness is of paramount importance in understanding the long-term implications of clinical resistance. Here we report the development of a novel recombinant virus assay to study entry inhibitor-resistant HIV variants using a biologically relevant cell type, primary CD4 T-cells. We have modified the replication-competent molecular clone HIV(NL4-3) to express a reporter protein (Renilla luciferase), Green Fluorescent Protein (EGFP), or Red Fluorescent Protein (DsRed2) upon infection, thus allowing quantification of replication. Luciferase-expressing virus was used to evaluate drug sensitivity, while co-infection with viruses carrying the green and red fluorescent proteins was employed in the competitive fitness assay. Using envelope proteins from three T20 insensitive variants, lower levels of resistance were observed in primary CD4 T-cells than had been previously reported for cell lines. Importantly, dual-color competition assays demonstrated comparable or higher fitness for these variants despite their reduced T20 sensitivity. We conclude that reduced sensitivity to T20 is compatible with high viral fitness in the absence of selection pressure. Thus, simultaneously measuring both resistance and viral fitness using this newly described dual-color competition assay will likely provide important information about resistant viral variants that emerge during therapy with entry inhibitors.
- Subjects :
- Amino Acid Sequence
CD4-Positive T-Lymphocytes drug effects
CD4-Positive T-Lymphocytes virology
Cell Line
Drug Resistance, Viral
Enfuvirtide
HIV Envelope Protein gp41 genetics
HIV Infections drug therapy
HIV Infections virology
HIV-1 isolation & purification
HIV-1 pathogenicity
HIV-1 physiology
Humans
Kinetics
Molecular Sequence Data
Sequence Homology, Amino Acid
Virus Replication drug effects
HIV Envelope Protein gp41 pharmacology
HIV Fusion Inhibitors pharmacology
HIV-1 drug effects
Microbial Sensitivity Tests methods
Peptide Fragments pharmacology
Subjects
Details
- Language :
- English
- ISSN :
- 0042-6822
- Volume :
- 333
- Issue :
- 2
- Database :
- MEDLINE
- Journal :
- Virology
- Publication Type :
- Academic Journal
- Accession number :
- 15721359
- Full Text :
- https://doi.org/10.1016/j.virol.2004.12.035