Inhibition of HIV-1 replication by novel lentiviral vectors expressing transdominant Rev and HIV-1 env antisense

Mr, Mautino; Ra, Morgan

doi:10.1038/sj.gt.3301674

Cited by 19 publications

(13 citation statements)

References 54 publications

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“…Challenges to bringing this therapy to the clinic include a difficulty in reaching efficacy, and the first HIV gene therapy clinical trial using modified T-cell transfer demonstrated the safety of the approach but did not achieve efficacy (9). Expression of antisense from lentivirusderived vectors offers increased transduction and efficacy in inhibiting HIV (25). Our vector is an HIV-based lentiviral vector expressing a long antisense for increased transduction and anti-HIV efficacy.…”

Section: Discussionmentioning

confidence: 99%

Antisense-Mediated Inhibition of Human Immunodeficiency Virus (HIV) Replication by Use of an HIV Type 1-Based Vector Results in Severely Attenuated Mutants Incapable of Developing Resistance

Lü

Qiao

Binder

et al. 2004

J Virol

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We have constructed a human immunodeficiency virus type 1 (HIV-1)-based lentiviral vector expressing a 937-base antisense sequence against the HIV-1 envelope gene. Transduction of CD4 ؉ T lymphocytes with this vector results in expression of the therapeutic antisense sequence and subsequent inhibition of productive HIV-1 replication. In this report, we examined the effect of antisense-mediated suppression on the potential development of virus escape mutants using a permissive T-cell line cultured under conditions that over serial passages specifically allowed for generation and amplification of mutants selected for by antisense pressure. In the resulting virus clones, we found a significant increase in the number of deletions at the envelope target region (91% compared to 27.5% in wild-type HIV). Deletions were most often greater than 1 kb in length. These data demonstrate for the first time that during antisense-mediated suppression of HIV, mutants develop as a direct result of selective pressure on the HIV genomic RNA. Interestingly, in clones where deletions were not observed, there was a high rate of A-G transitions in mutants at the antisense target region but not outside this region, which is consistent with those mutations that are predicted as a result of antisense-mediated modification of double-stranded RNA by the enzyme double-stranded RNA-specific adenosine deaminase. These clones were not found to be escape mutants, as their replicative ability was severely attenuated, and they did not replicate in the presence of vector.

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Section: Discussionmentioning

confidence: 99%

Antisense-Mediated Inhibition of Human Immunodeficiency Virus (HIV) Replication by Use of an HIV Type 1-Based Vector Results in Severely Attenuated Mutants Incapable of Developing Resistance

Lü

Qiao

Binder

et al. 2004

J Virol

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“…High-titer vesicular stomatitis virus-G pseudotyped retroviral supernatants concentrated 50Â by ultracentrifugation (20) were used to infect the PT-67 cell line (Clontech, Palo Alto, CA). High virus titer PT-67 clones with high GFP expression evaluated using a PhosphorImager (Molecular Dynamics, Sunnyvale, CA) and supernatants from these clones were used to transduce the human melanoma cell line 888 (21) to provide a relative estimate of retroviral titers (22). One clone that produced the GFP expressing virus (MIG) with a titer of 2 Â 10 8 transduction units and one that produced the GFP and Bcl-2 expressing virus (MIG-Bcl) with a titer of 4 Â 10 8 transduction units were selected for producing viruses for T-cell transduction.…”

Section: Methodsmentioning

confidence: 99%

Bcl-2 Overexpression Enhances Tumor-Specific T-Cell Survival

et al. 2005

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Although immunotherapy based on the adoptive transfer of tumor-specific T lymphocytes has been shown to result in dramatic clinical responses in some patients, the relatively low levels of engraftment and persistence of the adoptively transferred cells may limit these responses in many patients. In an attempt to develop strategies for prolonging the survival of adoptively transferred T cells, we have carried out studies in which T cells obtained from healthy donors as well as tumor-specific T cells were transduced with a retrovirus expressing the human Bcl-2 gene. Our results indicate that these transduced T cells overexpress Bcl-2, are resistant to death, and have a survival advantage following interleukin-2 withdrawal compared with control T cells transduced with a retrovirus expressing green fluorescent protein. Tumor-specific T cells overexpressing Bcl-2 maintained their ability to specifically recognize and respond to target cells. Furthermore, we show that adoptive immunotherapy of an established B16 tumor can be significantly enhanced by overexpressing Bcl-2 in melanoma-specific T-cell receptor transgenic T cells. Our data suggest that adoptive immunotherapy approaches to the treatment of cancer patients may be enhanced using Bcl-2-modified tumorreactive T cells. (Cancer Res 2005; 65(5): 2001-8)

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“…Among the RNA-based anti-HIV genes include a ribozyme that cleaves the U5 region of HIV-1 RNA by its enzymatic activity (Dropulic et al, 1996), an antisense RNA that hybridizes the transcripts of HIV-1 env gene to inhibit translation of Env (Mautino & Morgan, 2002a, c), and small interfering RNA (siRNA) that induces sequence-specific degradation of HIV-1 RNA (Banerjea et al, 2003). In the protein-based approach, the transdominant negative mutant of Rev (TdRev) is best described as an anti-HIV gene used in the setting of lentiviral vectors (Klimatcheva et al, 2001;Mautino et al, 2001;Mautino & Morgan, 2002c;Mukhtar et al, 2000). The TdReV named RevM10 is a derivative of HIV-1 Rev in which two amino acid mutations are introduced in the C-terminus activation domain, and hampers nuclear export of HIV-1 mRNAs via the formation of inactive multimers with WT Rev (Hope et al, 1992;Malim et al, 1989).…”

Section: Application Of a Gene Regulatable Lentiviral Vector For Hiv-mentioning

confidence: 99%

“…In this regard, the lentiviral vector has the potential advantage for transduction because of its ability to infect quiescent cells including HSC (Miyoshi et al, 1999). However, incorporation of anti-HIV genes into an HIV-based lentiviral vector can create problems for production of the vector itself; expression of the anti-HIV trans gene in the producer cells can interfere with vector production (Banerjea et al, 2003;Li et al, 2003;Mautino & Morgan, 2002c). One way to avoid this difficulty is to introduce a gene regulatable system in which the target transgene is kept silent during vector production and expression is subsequently induced on following infection of the vector in target cells.…”

Section: Introductionmentioning

confidence: 99%