Accumulation and Intranuclear Distribution of Unintegrated Human Immunodeficiency Virus Type 1 DNA

Bell, P. R.; Montaner, Luis J.; Maul, Gerd G.

doi:10.1128/jvi.75.16.7683-7691.2001

Cited by 61 publications

(54 citation statements)

References 62 publications

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“…However, while attachment to the primary target (i.e., carrier) cell is not pseudotype restricted, the susceptibility of secondary tissue targets remains subject to the specific interaction of vector envelope protein with its cognate cell surface receptor. Thus, for in vivo applications of vectorexposed cells, the combination of extended VSV-G tissue tropism and the ability of lentivirus vectors to transduce nondividing cells may lead to off-target transduction (3,22,37). Recent promising Env-based targeting approaches conferring tissue-restricted entry may be suitable in reducing off-target transduction in secondary tissues (41).…”

Section: Discussionmentioning

confidence: 99%

Prolonged Adherence of Human Immunodeficiency Virus-Derived Vector Particles to Hematopoietic Target Cells Leads to Secondary Transduction In Vitro and In Vivo

Pan¹,

Scarlett²,

Luoh³

et al. 2007

J Virol

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Human immunodeficiency virus type 1-derived lentivirus vectors bearing the vesicular stomatitis virus G (VSV-G) envelope glycoprotein demonstrate a wide host range and can stably transduce quiescent hematopoietic stem cells. In light of concerns about biosafety and potential germ line transmission, they have been used predominantly for ex vivo strategies, thought to ensure the removal of excess surface-bound particles and prevent in vivo dissemination. Studies presented here instead reveal prolonged particle adherence after ex vivo exposure, despite serial wash procedures, with subsequent transduction of secondary target cells in direct and transwell cocultures. We explored the critical parameters affecting particle retention and transfer and show that attachment to the cell surface selectively protects virus particles from serum complement-mediated inactivation. Moreover, studies with nonmyeloablated murine recipients show that transplantation of vectorexposed, washed hematopoietic cells results in systemic dissemination of functional VSV-G/lentivector particles. We demonstrate genetic marking by inadvertent transfer of vector particles and prolonged expression of transgene product in recipient tissues. Our findings have implications for biosafety, vector design, and cell biology research.

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Section: Discussionmentioning

confidence: 99%

Prolonged Adherence of Human Immunodeficiency Virus-Derived Vector Particles to Hematopoietic Target Cells Leads to Secondary Transduction In Vitro and In Vivo

Pan¹,

Scarlett²,

Luoh³

et al. 2007

J Virol

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Section: Rna Virusesmentioning

confidence: 99%

“…Bell et al [125] reported that no significant relationship was observed between ND10 or any of the following: HIV-1 DNA, transcription foci, and integrated DNA. Their results showed that HIV-1 did not modify ND10 at early or late times of infection [125] . However, Turelli et al [126] reported that incoming retroviral preintegration complexes trigger the exportingmediated cytoplasmic export of PML.…”

Section: Rna Virusesmentioning

confidence: 99%

Nuclear domain 10 of the viral aspect

Rivera-Molina

Martínez²,

Tang³

2013

WJV

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Nuclear domain 10 (ND10) are spherical bodies distributed throughout the nucleoplasm and measuring around 0.2-1.0 μm. First observed under an electron microscope, they were originally described as dense bodies found in the nucleus. They are known by a number of other names, including Promyelocytic Leukemia bodies (PML bodies), Kremer bodies, and PML oncogenic domains. ND10 are frequently associated with Cajal bodies and cleavage bodies. It has been suggested that they play a role in regulating gene transcription. ND10 were originally characterized using human autoantisera, which recognizes Speckled Protein of 100 kDa, from patients with primary biliary cirrhosis. At the immunohistochemical level, ND10 appear as nuclear punctate structures, with 10 indicating the approximate number of dots per nucleus observed. ND10 do not colocalize with kinetochores, centromeres, sites of mRNA processing, or chromosomes. Resistance of ND10 antigens to nuclease digestion and salt extraction suggest that ND10 are associated with the nuclear matrix.They are often identified by immunofluorescent assay using specific antibodies against PML, Death domainassociated protein, nuclear dot protein (NDP55), and so on. The role of ND10 has long been the subject of investigation, with the specific connection of ND10 and viral infection having been a particular focus for almost 20 years. This review summarizes the relationship of ND10 and viral infection. Some future study directions are also discussed.

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“…Even though FISH is a powerful technique, it is not very sensitive for HIV-1 detection and moreover disrupts the native architecture of the nuclear compartment as it requires harsh denaturation conditions. In addition, this technique does not allow the discrimination between integrated and nonintegrated viral DNA (16,17). Here we describe a fluorescent approach to visualize HIV-1 DNA in the nuclear compartment of infected cells.…”

mentioning

confidence: 99%

Single-Cell Imaging of HIV-1 Provirus (SCIP)

Primio

Quercioli

Allouch

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

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Recent advances in fluorescence microscopy provided tools for the investigation and the analysis of the viral replication steps in the cellular context. In the HIV field, the current visualization systems successfully achieve the fluorescent labeling of the viral envelope and proteins, but not the genome. Here, we developed a system able to visualize the proviral DNA of HIV-1 through immunofluorescence detection of repair foci for DNA double-strand breaks specifically induced in the viral genome by the heterologous expression of the I-SceI endonuclease. The system for Single-Cell Imaging of HIV-1 Provirus, named SCIP, provides the possibility to individually track integrated-viral DNA within the nuclei of infected cells. In particular, SCIP allowed us to perform a topological analysis of integrated viral DNA revealing that HIV-1 preferentially integrates in the chromatin localized at the periphery of the nuclei.T echnical developments in imaging-based techniques have greatly improved our understanding of HIV-host cell interactions. HIV-1 virions labeled with fluorophores were pivotal in shedding light onto multiple aspects of the virus-host interplay during all steps of HIV-1 replication cycle (1-13). Nevertheless, few optical approaches have been so far developed to visualize viral particles within the nuclear compartment (14, 15), which limits our comprehension of the interaction between HIV-1 and the nuclear architecture. Moreover, the existing detection tools are based on the visualization of the viral protein complexes or envelope but not of the viral DNA with the only exception of the fluorescence in situ hybridization (FISH) technique. Even though FISH is a powerful technique, it is not very sensitive for HIV-1 detection and moreover disrupts the native architecture of the nuclear compartment as it requires harsh denaturation conditions. In addition, this technique does not allow the discrimination between integrated and nonintegrated viral DNA (16, 17). Here we describe a fluorescent approach to visualize HIV-1 DNA in the nuclear compartment of infected cells. We exploited a site-specific genome engineering technique that represents one of the most promising approaches to detect specific genome regions in modified organism (18) allowing for their spatial localization in the cell (19,20). This technique couples endogenous repair pathways, induced by rare cutting endonuclease, with immunofluorescence analysis. Rare cutting endonucleases, such as the yeast-homing endonuclease I-SceI, specifically cuts target sequences that cannot be found in the mammalian genome. By engineering DNA to contain the I-SceI cleavage site, it is thus possible to induce endogenous repair mechanism for double-strand breaks (DSBs) at specific genomic positions. DSB repair leads to the formation of distinct subnuclear structures that are generally referred to as "foci" (21). The first sensor of the DSB is the histone H2AX, which becomes massively phosphorylated at serine 139 (γ-H2AX). Foci of DNA repair are thus visible through immu...

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Accumulation and Intranuclear Distribution of Unintegrated Human Immunodeficiency Virus Type 1 DNA

Cited by 61 publications

References 62 publications

Prolonged Adherence of Human Immunodeficiency Virus-Derived Vector Particles to Hematopoietic Target Cells Leads to Secondary Transduction In Vitro and In Vivo

Prolonged Adherence of Human Immunodeficiency Virus-Derived Vector Particles to Hematopoietic Target Cells Leads to Secondary Transduction In Vitro and In Vivo

Nuclear domain 10 of the viral aspect

Single-Cell Imaging of HIV-1 Provirus (SCIP)

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