Abstract

This application proposes to do a detailed study of the phenotypic, genotypic, virologic, and biochemical characteristics of HIV-1 isolates that are dually resistant to AZT and 3TC. Susceptibility of HIV-1 to AZT and 3TC will be tested with a PBMC assay as well as a recombinant virus assay using low-passage isolates and biologically cloned virus. The nucleotide sequence of full-length molecular clones of RT from dually resistant strains will be determined and compared to AZT-sensitive strains. The association of specific mutations with AZT phenotype will be examined by a linear mixed-effects model. Candidate point mutations will be confirmed by in vitro mutagenesis and phenotypic analysis of the resulting recombinant viruses. The growth kinetics of dually resistant viruses will also be compared to wild-type and singlely resistant isolates. To characterize the biochemical properties of dually resistant RTs, the genes encoding the p66 and p51 RT subunits will be cloned separately into E. coli expression vectors and purified to near homogeneity. Mechanistic studies with reconstituted p66/p51 heterodimers will be performed using synthetic homopolymeric, as well as native heteromeric, RNA templates.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI042567-03
Application #
6137253
Study Section
AIDS and Related Research Study Section 4 (ARRD)
Program Officer
Bridges, Sandra H
Project Start
1998-01-01
Project End
2001-08-14
Budget Start
2000-01-01
Budget End
2001-08-14
Support Year
3
Fiscal Year
2000
Total Cost
$273,688
Indirect Cost

  Institution

Name
University of Colorado Denver
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
065391526
City
Aurora
State
CO
Country
United States
Zip Code
80045

  Publications

Paredes, Roger; Sagar, Manish; Marconi, Vincent C et al. (2009) In vivo fitness cost of the M184V mutation in multidrug-resistant human immunodeficiency virus type 1 in the absence of lamivudine. J Virol 83:2038-43
Paredes, Roger; Cheng, Irene; Kuritzkes, Daniel R et al. (2007) High prevalence of primary lamivudine and nelfinavir resistance in HIV-1-infected pregnant women in the United States, 1998-2004. AIDS 21:2103-6
Hu, Zixin; Hatano, Hiroyu; Hammond, Sarah P et al. (2007) Virologic characterization of HIV type 1 with a codon 70 deletion in reverse transcriptase. J Acquir Immune Defic Syndr 45:494-500
Hu, Zixin; Giguel, Francoise; Hatano, Hiroyu et al. (2006) Fitness comparison of thymidine analog resistance pathways in human immunodeficiency virus type 1. J Virol 80:7020-7
Campbell, Thomas B; Shulman, Nancy S; Johnson, Steven C et al. (2005) Antiviral activity of lamivudine in salvage therapy for multidrug-resistant HIV-1 infection. Clin Infect Dis 41:236-42
Lu, Jing; Whitcomb, Jeannette; Kuritzkes, Daniel R (2005) Effect of the Q207D mutation in HIV type 1 reverse transcriptase on zidovudine susceptibility and replicative fitness. J Acquir Immune Defic Syndr 40:20-3
Miranda, Luis R; Gotte, Matthias; Liang, Fei et al. (2005) The L74V mutation in human immunodeficiency virus type 1 reverse transcriptase counteracts enhanced excision of zidovudine monophosphate associated with thymidine analog resistance mutations. Antimicrob Agents Chemother 49:2648-56
Kuritzkes, Daniel R; Bassett, Roland L; Hazelwood, J Darren et al. (2004) Rate of thymidine analogue resistance mutation accumulation with zidovudine- or stavudine-based regimens. J Acquir Immune Defic Syndr 36:600-3
Stoeckli, Thomas C; MaWhinney, Samantha; Uy, Jonathan et al. (2002) Phenotypic and genotypic analysis of biologically cloned human immunodeficiency virus type 1 isolates from patients treated with zidovudine and lamivudine. Antimicrob Agents Chemother 46:4000-3
Brown, E R; MaWhinney, S; Jones, R H et al. (2001) Improving the fit of bivariate smoothing splines when estimating longitudinal immunological and virological markers in HIV patients with individual antiretroviral treatment strategies. Stat Med 20:2489-504

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