Inhibition of HIV‐1 replication in primary human T cells transduced with an intracellular anti‐HIV‐1 p17 antibody gene

Tewari, Deepanker; Notkins, Abner Louis; Zhou, Paul

doi:10.1002/jgm.336

Cited by 14 publications

(9 citation statements)

References 41 publications

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“…In this study, we demonstrated that the expression of 2LTRZFP-GFP, in the nucleus of human T-lymphocytic cells, markedly inhibited viral replication and integration as measured by the p24 antigen assay and Alu-gag qPCR, respectively. The HIV-1 integrase recognition sequence is highly conserved and is processed by HIV-1 IN in the early phase of viral replication (LaFemina et al, 1991;Vink et al, 1991;van den Ent et al, 1994;Yoshinaga and Fujiwara, 1995;Katzman and Sudol, 1996;Balakrishnan and Jonsson, 1997). Although certain integrase inhibitors are currently being approved and used for the treatment of HIV-1-infected FIG.…”

Section: Discussionmentioning

confidence: 99%

“…However, RNA approaches may be limited by escape mutations in the site-targeting regions, even without the changing of encoded protein. Many protein interference strategies involve trans-dominant negative mutants (Liem et al, 1993) and single-chain variable fragments (scFvs) to inhibit the function of intracellular target proteins in specific cellular compartments, such as IN (Levy-Mintz et al, 1996), reverse transcriptase (RT) (Shaheen et al, 1996), Rev (Vercruysse et al, 2010), gp120 (Marasco et al, 1993), and Gag p17 (Tewari et al, 2003). However, the reducing environment in the cytosol is an obstacle to the formation of disulfide bonds required for scFv function.…”

Section: Introductionmentioning

confidence: 99%

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Zinc Finger Protein Designed to Target 2-Long Terminal Repeat Junctions Interferes with Human Immunodeficiency Virus Integration

et al. 2012

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Integration of the human immunodeficiency virus type 1 (HIV-1) genome into the host chromosome is a vital step in the HIV life cycle. The highly conserved cytosine-adenine (CA) dinucleotide sequence immediately upstream of the cleavage site is crucial for integrase (IN) activity. As this viral enzyme has an important role early in the HIV-1 replication cycle, interference with the IN substrate has become an attractive strategy for therapeutic intervention. We demonstrated that a designed zinc finger protein (ZFP) fused to green fluorescent protein (GFP) targets the 2-long terminal repeat (2-LTR) circle junctions of HIV-1 DNA with nanomolar affinity. We report now that 2LTRZFP-GFP stably transduced into 293T cells interfered with the expression of vesicular stomatitis virus glycoprotein (VSV-G)-pseudotyped lentiviral red fluorescent protein (RFP), as shown by the suppression of RFP expression. We also used a third-generation lentiviral vector and pCEP4 expression vector to deliver the 2LTRZFP-GFP transgene into human T-lymphocytic cells, and a stable cell line for long-term expression studies was selected for HIV-1 challenge. HIV-1 integration and replication were inhibited as measured by Alu-gag real-time PCR and p24 antigen assay. In addition, the molecular activity of 2LTRZFP-GFP was evaluated in peripheral blood mononuclear cells. The results were confirmed by Alu-gag real-time PCR for integration interference. We suggest that the expression of 2LTRZFP-GFP limited viral integration on intracellular immunization, and that it has potential for use in HIV gene therapy in the future.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Zinc Finger Protein Designed to Target 2-Long Terminal Repeat Junctions Interferes with Human Immunodeficiency Virus Integration

et al. 2012

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“…24 In another example, retroviral plasmid-based delivery was used to express intrabodies to the a-HIV-1 Gag p17 protein in the cytoplasm of primary human T cells and in Jurkat cells (a human CD4(+) T cell line). 25 When these cells were challenged with several strains of HIV-1, replication was strongly inhibited, although the degree of resistance varied and was not efficient in cells that sustained high viral replication. The inhibition was at post-entry and reverse transcription levels, as only p24 and not proviral DNA differed between the scFvtransduced human primary T cells and control.…”

Section: Intrabodies Against Viral or Viral-related Host Proteins Canmentioning

confidence: 99%

Gene therapy progress and prospects: Novel gene therapy approaches for AIDS

Wolkowicz

Nolan

2005

Gene Ther

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“…To date, there are no available data about the effect of nuclear scFv fragments in viral potyvirus infections, although the potential of nuclear scFvs to interfere with HIV replication function has been reported (Sibler et al. , 2003; Tewari et al. , 2003).…”

Section: Discussionmentioning

confidence: 99%

Resistance to Plum pox virus in plants expressing cytosolic and nuclear single‐chain antibodies against the viral RNA NIb replicase

Gil¹,

Esteban²,

Garcı́a³

et al. 2011

Plant Pathology

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The expression of engineered single-chain variable fragments specific to the NIb RNA replicase of Plum pox virus (PPV) (scFv2A) in transgenic plants was successfully used as a strategy to interfere with viral infection. Different scFv2A fusion proteins were constructed to target those subcellular compartments, such as the cytosol, endoplasmic reticulum (ER) membrane structures and the nucleus, where NIb protein presumably accumulates. Several transgenic lines of Nicotiana benthamiana plants expressing the scFv2A targeted to the cytosol (2A lines), ER (6K2 lines) and nucleus (NLS lines) were obtained. The protective effect of scFv expression was determined by mechanical virus inoculation in five 2A, three 6K2 and four NLS transgenic lines. The strongest resistance was afforded with the 2A-3 (six non-infected plants out of 10), 6K2-1 (17 out of 33) and NLS-11 (16 out of 19) transgenic lines. The success of this interference with PPV infection opens new possibilities for the control of this RNA virus and could be exploited not only to confer resistance in transgenic plants, but also to elucidate the role of the non-structural NIb protein in different cell compartments during viral infection.

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Inhibition of HIV‐1 replication in primary human T cells transduced with an intracellular anti‐HIV‐1 p17 antibody gene

Abstract: The expression of the anti-HIV-1 gag p17 scFv/Ckappa gene construct in primary human T cells renders these cells resistant to HIV-1 and points to the potential clinical usefulness of this gene construct for anti-HIV-1 gene therapy.

Cited by 14 publications

References 41 publications

Zinc Finger Protein Designed to Target 2-Long Terminal Repeat Junctions Interferes with Human Immunodeficiency Virus Integration

Zinc Finger Protein Designed to Target 2-Long Terminal Repeat Junctions Interferes with Human Immunodeficiency Virus Integration

Gene therapy progress and prospects: Novel gene therapy approaches for AIDS

Resistance to Plum pox virus in plants expressing cytosolic and nuclear single‐chain antibodies against the viral RNA NIb replicase

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