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

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, and Robert J. Suhadolnik^1,2,*

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-)-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  subunit of eukaryotic initiation factor 2 and NF-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--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.

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:

• 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]  
• MOSOIAN, A., TEIXEIRA, A., BURNS, C. S., KHITROV, G., ZHANG, W., GUSELLA, L., KLOTMAN, P., KLOTMAN, M. (2007). Influence of Prothymosin-{alpha} on HIV-1 Target Cells. Ann. N. Y. Acad. Sci. 1112: 269-285 [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]