Cell-type-specific gene delivery into neuronal cells in vitro and in vivo

Parveen, Zahida; Mukhtār, Muḥammad; Rafi, M A; Wenger, David A.; Siddiqui, Khwaja M.; Siler, Catherine A.; Dietzschold, Bernhard; Pomerantz, Roger J.; Schnell, Matthias J.; Dornburg, Alex

doi:10.1016/s0042-6822(03)00402-1

Cited by 16 publications

(17 citation statements)

References 36 publications

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“…The SNV-Nef vector was generated by cloning the nef open reading frame into the SNV transfer vector pZP35 via blunt-ended ligation (78,79). The Nef fragment was generated as indicated above, and the pZP35 was linearized with SmaI restriction endonuclease.…”

Section: Methodsmentioning

confidence: 99%

Human Immunodeficiency Virus Type 1 Nef Potently Induces Apoptosis in Primary Human Brain Microvascular Endothelial Cells via the Activation of Caspases

Acheampong

Parveen

Muthoga

et al. 2005

J Virol

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The lentiviral protein Nef plays a major role in the pathogenesis of human immunodeficiency virus type I (HIV-1) infection. Although the exact mechanisms of its actions are not fully understood, Nef has been shown to be essential for the maintenance of high-titer viral replication and disease pathogenesis in in vivo models of simian immunodeficiency virus infection of monkeys. Nef has also been suggested to play a pivotal role in the depletion of T cells by promoting apoptosis in bystander cells. In this context, we investigated the ability of extracellular and endogenously expressed HIV-1 Nef to induce apoptosis in primary human brain microvascular endothelial cells (MVECs). Human brain MVECs were exposed to baculovirus-expressed HIV-1 Nef protein, an HIV-1-based vector expressing Nef, spleen necrosis virus (SNV)-Nef virus (i.e., SNV vector expressing HIV-1 Nef as a transgene), and the HIV-1 strain ADA and its Nef deletion mutant, ADA⌬Nef. We observed that ADA Nef, the HIV-1 vector expressing Nef, and SNV-Nef were able to induce apoptosis in a dose-dependent manner. The mutant virus with a deletion in Nef was able to induce apoptosis in MVECs to modest levels, but the effects were not as pronounced as with the wild-type HIV-1 strain, ADA, the HIV-1-based vector expressing Nef, or SNV-Nef viruses. We also demonstrated that relatively high concentrations of exogenous HIV-1 Nef protein were able to induce apoptosis in MVECs. Gene microarray analyses showed increases in the expression of several specific proapoptotic genes. Western blot analyses revealed that the various caspases involved with Nef-induced apoptosis are processed into cleavage products, which occur only during programmed cell death. The results of this study demonstrate that Nef likely contributes to the neuroinvasion and neuropathogenesis of HIV-1, through its effects on select cellular processes, including various apoptotic cascades.

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Section: Methodsmentioning

confidence: 99%

Human Immunodeficiency Virus Type 1 Nef Potently Induces Apoptosis in Primary Human Brain Microvascular Endothelial Cells via the Activation of Caspases

Acheampong

Parveen

Muthoga

et al. 2005

J Virol

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“…The predominant hypothesis is that, unlike HIV, MLV lacks a NLS required for active nuclear import (34). The presence of a NLS has also been implicated in the in vitro infection of nondividing cells by alpharetroviruses, but this has not been confirmed in vivo (24,26,30,31,44). However, previous MLV variants with NLSs engineered at fixed locations in IN or proviral DNA were unable to infect growtharrested cells (35,48).…”

Section: Discussionmentioning

confidence: 99%

High-Throughput, Library-Based Selection of a Murine Leukemia Virus Variant To Infect Nondividing Cells

Schaffer

2006

J Virol

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“…Retroviral vector pZP 32 is an SNV-based REV A (Gag) packaging construct. The SNV gag (334 bp) was replaced by the REV A gag (342 bp) with nuclear localization signal sequences within the SNV pol, as described previously (38,39) (Fig. 1A).…”

Section: Methodsmentioning

confidence: 99%

“…1A) was derived from pAK3 (derived from pO11) by the addition of the CMV promoter at the XhoI site in the U3 region of the 3Ј LTR through a XhoI linker (39). The ␤-galactosidase (␤-Gal) gene was cloned as a marker gene by using XbaI and HindIII sites (Fig.…”

Section: Methodsmentioning

confidence: 99%

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Cross-Packaging of Human Immunodeficiency Virus Type 1 Vector RNA by Spleen Necrosis Virus Proteins: Construction of a New Generation of Spleen Necrosis Virus-Derived Retroviral Vectors

Parveen¹,

Mukhtār²,

Goodrich

et al. 2004

J Virol

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The ability of the nonlentiviral retrovirus spleen necrosis virus (SNV) to cross-package the genomic RNA of the distantly related human immunodeficiency virus type 1 (HIV-1) and vice versa was analyzed. Such a model may allow us to further study HIV-1 replication and pathogenesis, as well as to develop safe gene therapy vectors. Our results suggest that SNV can cross-package HIV-1 genomic RNA but with lower efficiency than HIV-1 proteins. However, HIV-1-specific proteins were unable to cross-package SNV RNA. We also constructed SNV-based gag-pol chimeric variants by replacing the SNV integrase with the HIV-1 integrase, based on multiple sequence alignments and domain analyses. These analyses revealed that there are conserved domains in all retroviral integrase open reading frames (orf), despite the divergence in the primary sequences. The transcomplementation assays suggested that SNV proteins recognized one of the chimeric variants. This demonstrated that HIV-1 integrase is functional in the SNV gag-pol orf with a lower transduction efficiency, utilizing homologous (SNV) RNA, as well as the heterologous vector RNA of HIV-1. These findings suggest that homology in the conserved sequences of the integrase protein may not be fully competent in the replacement of protein(s) from one retrovirus to another, and there are likely several other factors involved in each of the steps related to replication, integration, and infection. However, further studies to dissect the gag-pol region will be critical for understanding the mechanisms involved in the cleavage of reverse transcriptase, RNase H, and integrase. These studies should provide further insight into the design and development of novel molecular approaches to block HIV-1 replication and to construct a new generation of SNV-based vectors.

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Cell-type-specific gene delivery into neuronal cells in vitro and in vivo

Cited by 16 publications

References 36 publications

Human Immunodeficiency Virus Type 1 Nef Potently Induces Apoptosis in Primary Human Brain Microvascular Endothelial Cells via the Activation of Caspases

Human Immunodeficiency Virus Type 1 Nef Potently Induces Apoptosis in Primary Human Brain Microvascular Endothelial Cells via the Activation of Caspases

High-Throughput, Library-Based Selection of a Murine Leukemia Virus Variant To Infect Nondividing Cells

Cross-Packaging of Human Immunodeficiency Virus Type 1 Vector RNA by Spleen Necrosis Virus Proteins: Construction of a New Generation of Spleen Necrosis Virus-Derived Retroviral Vectors

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