This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Muto, N. F.
Right arrow Articles by Suhadolnik, R. J.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Muto, N. F.
Right arrow Articles by Suhadolnik, R. J.

 Previous Article  |  Next Article 

Journal of Virology, November 1999, p. 9021-9028, Vol. 73, No. 11
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Inhibition of Replication of Reactivated Human Immunodeficiency Virus Type 1 (HIV-1) in Latently Infected U1 Cells Transduced with an HIV-1 Long Terminal Repeat-Driven PKR cDNA Construct

Nicholas F. Muto,1 Camille Martinand-Mari,2 Martin E. Adelson,1,dagger and Robert J. Suhadolnik1,2,*

Fels Institute for Cancer Research and Molecular Biology1 and Department of Biochemistry,2 Temple University School of Medicine, Philadelphia, Pennsylvania 19140

Received 14 April 1999/Accepted 6 August 1999

Treatment of human immunodeficiency virus type 1 (HIV-1)-infected individuals with highly active antiretroviral therapy has effectively decreased viral load to undetectable levels. However, efforts to eliminate HIV-1 from these individuals have been unsuccessful, due to the presence of stable, latent viral reservoirs in resting and active CD4+ T lymphocytes and macrophages. These latent populations have become critical targets in the effort to eradicate HIV-1 from infected individuals. The mechanisms of HIV-1 latency have been studied by using the HIV-1-infected promonocytic cell line U1. The interferon-inducible double-stranded RNA-dependent p68 protein kinase (PKR), a key enzyme in the host-mediated antiviral response, is known to be down-regulated during HIV-1 infection. Therefore, in order to evaluate the role of PKR in the inhibition of replication of reactivated HIV-1 in latently infected U1 cells, we have utilized cDNA constructs containing PKR under the transcriptional control of the HIV-1 long terminal repeat. One PKR-transduced clone, U1/106-4:27, inhibited the tumor necrosis factor alpha (TNF-alpha )-induced replication of HIV-1 by 99% compared to control U1 cells as measured by syncytium formation and HIV-1 p24 antigen enzyme-linked immunosorbent assay. Western blot analysis showed an increase in PKR expression through 96 h postinduction in the U1/106-4:27 clone, concomitant with maximal increases in phosphorylation of the alpha  subunit of eukaryotic initiation factor 2 and NF-kappa B activity at 72 h postinduction. These results demonstrate that overexpression of PKR can inhibit the replication of reactivated HIV-1 in latently infected cells and confirm the involvement of PKR in the interferon-associated antiviral pathway against HIV-1 infection. Additionally, treatment of the PKR-transduced U1/106-4:27 clone with the protease inhibitor saquinavir (250 nM) completely inhibited TNF-alpha -induced HIV-1 replication.


* Corresponding author. Mailing address: Department of Biochemistry, Temple University School of Medicine, Philadelphia, PA 19140. Phone: (215) 707-4607. Fax: (215) 707-3515. E-mail: rjs{at}astro.ocis.temple.edu.

dagger Present address: Department of Molecular Genetics and Microbiology, UMDNJ-Robert Wood Johnson Medical School, Piscataway, NJ 08854.


Journal of Virology, November 1999, p. 9021-9028, Vol. 73, No. 11
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.



This article has been cited by other articles:

  • Hu, W., Hofstetter, W., Wei, X., Guo, W., Zhou, Y., Pataer, A., Li, H., Fang, B., Swisher, S. G. (2009). Double-Stranded RNA-Dependent Protein Kinase-Dependent Apoptosis Induction by a Novel Small Compound. J. Pharmacol. Exp. Ther. 328: 866-872 [Abstract] [Full Text]  
  • Goplen, N., Gorska, M. M., Stafford, S. J., Rozario, S., Guo, L., Liang, Q., Alam, R. (2008). A Phosphosite Screen Identifies Autocrine TGF-{beta}-Driven Activation of Protein Kinase R as a Survival-Limiting Factor for Eosinophils. J. Immunol. 180: 4256-4264 [Abstract] [Full Text]  
  • Ong, C. L., Thorpe, J. C., Gorry, P. R., Bannwarth, S., Jaworowski, A., Howard, J. L., Chung, S., Campbell, S., Christensen, H. S., Clerzius, G., Mouland, A. J., Gatignol, A., Purcell, D. F. J. (2005). Low TRBP Levels Support an Innate Human Immunodeficiency Virus Type 1 Resistance in Astrocytes by Enhancing the PKR Antiviral Response. J. Virol. 79: 12763-12772 [Abstract] [Full Text]  
  • Espert, L., Degols, G., Lin, Y.-L., Vincent, T., Benkirane, M., Mechti, N. (2005). Interferon-induced exonuclease ISG20 exhibits an antiviral activity against human immunodeficiency virus type 1. J. Gen. Virol. 86: 2221-2229 [Abstract] [Full Text]  
  • Rogez-Kreuz, C., Maneglier, B., Martin, M., Dereuddre-Bosquet, N., Martal, J., Dormont, D., Clayette, P. (2005). Involvement of IL-6 in the anti-human immunodeficiency virus activity of IFN-{tau} in human macrophages. Int Immunol 17: 1047-1057 [Abstract] [Full Text]  
  • Rogez, C., Martin, M., Dereuddre-Bosquet, N., Martal, J., Dormont, D., Clayette, P. (2003). Anti-Human Immunodeficiency Virus Activity of Tau Interferon in Human Macrophages: Involvement of Cellular Factors and {beta}-Chemokines. J. Virol. 77: 12914-12920 [Abstract] [Full Text]  
  • Kutsch, O., Benveniste, E. N., Shaw, G. M., Levy, D. N. (2002). Direct and Quantitative Single-Cell Analysis of Human Immunodeficiency Virus Type 1 Reactivation from Latency. J. Virol. 76: 8776-8786 [Abstract] [Full Text]  
  • Samuel, C. E. (2001). Antiviral Actions of Interferons. Clin. Microbiol. Rev. 14: 778-809 [Abstract] [Full Text]  
  • Lever, A M L (2001). Gene therapy for HIV. Sex. Transm. Infect. 77: 93-96 [Full Text]