scholarly journals Vesicular stomatitis virus enables gene transfer and transsynaptic tracing in a wide range of organisms

2015 ◽  
Vol 523 (11) ◽  
pp. 1639-1663 ◽  
Author(s):  
Nathan A. Mundell ◽  
Kevin T. Beier ◽  
Y. Albert Pan ◽  
Sylvain W. Lapan ◽  
Didem Göz Aytürk ◽  
...  
Keyword(s):  
Gene Transfer ◽  
Vesicular Stomatitis Virus ◽  
Wide Range ◽  
Vesicular Stomatitis ◽  
Transsynaptic Tracing

scholarly journals Phosphatidylserine Is Not the Cell Surface Receptor for Vesicular Stomatitis Virus

2004 ◽  
Vol 78 (20) ◽  
pp. 10920-10926 ◽  
Author(s):  
David A. Coil ◽  
A. Dusty Miller
Keyword(s):  
Cell Surface ◽  
Vesicular Stomatitis Virus ◽  
Virus Entry ◽  
Annexin V ◽  
Cell Types ◽  
Membrane Lipid ◽  
Cell Surface Receptor ◽  
Wide Range ◽  
Surface Receptor ◽  
Vesicular Stomatitis

ABSTRACT The envelope protein from vesicular stomatitis virus (VSV) has become an important tool for gene transfer and gene therapy. It is widely used mainly because of its ability to mediate virus entry into all cell types tested to date. Consistent with the broad tropism of the virus, the receptor for VSV is thought to be a ubiquitous membrane lipid, phosphatidylserine (PS). However, the evidence for this hypothesis is indirect and incomplete. Here, we have examined the potential interaction of VSV and PS at the plasma membrane in more detail. Measurements of cell surface levels of PS show a wide range across cell types from different organisms. We demonstrate that there is no correlation between the cell surface PS levels and VSV infection or binding. We also demonstrate that an excess of annexin V, which binds specifically and tightly to PS, does not inhibit infection or binding by VSV. While the addition of PS to cells does allow increased virus entry, we show that this effect is not specific to the VSV envelope. We conclude that PS is not the cell surface receptor for VSV, although it may be involved in a postbinding step of virus entry.

scholarly journals Vesicular Stomatitis Virus G-Pseudotyped Lentivirus Vectors Mediate Efficient Apical Transduction of Polarized Quiescent Primary Alveolar Epithelial Cells

2001 ◽  
Vol 75 (23) ◽  
pp. 11747-11754 ◽  
Author(s):  
Zea Borok ◽  
Jens Erik Harboe-Schmidt ◽  
Steven L. Brody ◽  
Yingjian You ◽  
Beiyun Zhou ◽  
...  
Keyword(s):  
Gene Transfer ◽  
Epithelial Cells ◽  
G Protein ◽  
Vesicular Stomatitis Virus ◽  
Apical Cell ◽  
Alveolar Epithelial Cells ◽  
Lentivirus Vector ◽  
Alveolar Epithelial ◽  
Vesicular Stomatitis ◽  
Lentivirus Vectors

ABSTRACT We investigated the use of lentivirus vectors for gene transfer to quiescent alveolar epithelial cells. Primary rat alveolar epithelial cells (AEC) grown on plastic or as polarized monolayers on tissue culture-treated polycarbonate semipermeable supports were transduced with a replication-defective human immunodeficiency virus-based lentivirus vector pseudotyped with the vesicular stomatitis virus G (VSV-G) protein and encoding an enhanced green fluorescent protein reporter gene. Transduction efficiency, evaluated by confocal microscopy and quantified by fluorescence-activated cell sorting, was dependent on the dose of vector, ranging from 4% at a multiplicity of infection (MOI) of 0.1 to 99% at an MOI of 50 for AEC grown on plastic. At a comparable titer and MOI, transduction of these cells by a similarly pseudotyped murine leukemia virus vector was ∼30-fold less than by the lentivirus vector. Importantly, comparison of lentivirus-mediated gene transfer from the apical or basolateral surface of confluent AEC monolayers (Rt > 2 kΩ · cm2; MOI = 10) revealed efficient transduction only when VSV-G-pseudotyped lentivirus was applied apically. Furthermore, treatment with EGTA to increase access to the basolateral surface did not increase transduction of apically applied virus, indicating that transduction was primarily via the apical membrane domain. In contrast, differentiated tracheal epithelial cells were transduced by apically applied lentivirus only in the presence of EGTA and at a much lower overall efficiency (∼15-fold) than was observed for AEC. Efficient transduction of AEC from the apical cell surface supports the feasibility of using VSV-G-pseudotyped lentivirus vectors for gene transfer to the alveolar epithelium and suggests that differences exist between upper and lower airways in the polarity of available receptors for the VSV-G protein.

scholarly journals Cholesterol 25-hydroxylase suppresses SARS-CoV-2 replication by blocking membrane fusion

2020 ◽  
Vol 117 (50) ◽  
pp. 32105-32113 ◽  
Author(s):  
Ruochen Zang ◽  
James Brett Case ◽  
Eylan Yutuc ◽  
Xiucui Ma ◽  
Sheng Shen ◽  
...  
Keyword(s):  
Membrane Fusion ◽  
Vesicular Stomatitis Virus ◽  
Molecular Basis ◽  
Enveloped Viruses ◽  
Late Endosomes ◽  
Wide Range ◽  
Therapeutic Development ◽  
Antiviral Activities ◽  
Vesicular Stomatitis ◽  
Catalyzed Membrane

Cholesterol 25-hydroxylase (CH25H) is an interferon (IFN)-stimulated gene that shows broad antiviral activities against a wide range of enveloped viruses. Here, using an IFN-stimulated gene screen against vesicular stomatitis virus (VSV)-SARS-CoV and VSV-SARS-CoV-2 chimeric viruses, we identified CH25H and its enzymatic product 25-hydroxycholesterol (25HC) as potent inhibitors of SARS-CoV-2 replication. Internalized 25HC accumulates in the late endosomes and potentially restricts SARS-CoV-2 spike protein catalyzed membrane fusion via blockade of cholesterol export. Our results highlight one of the possible antiviral mechanisms of 25HC and provide the molecular basis for its therapeutic development.

scholarly journals Enhanced Gene Transfer with Fusogenic Liposomes Containing Vesicular Stomatitis Virus G Glycoprotein

1998 ◽  
Vol 72 (7) ◽  
pp. 6159-6163 ◽  
Author(s):  
Akihiro Abe ◽  
Atsushi Miyanohara ◽  
Theodore Friedmann
Keyword(s):  
Gene Transfer ◽  
Vesicular Stomatitis Virus ◽  
Sucrose Density Gradient ◽  
Virus Envelope ◽  
Lipid Complex ◽  
Targeted Gene Delivery ◽  
Vesicular Stomatitis ◽  
Sucrose Density Gradient Sedimentation

ABSTRACT Exposure of Lipofectin-DNA complexes to the partially purified G glycoprotein of the vesicular stomatitis virus envelope (VSV-G) results in loss of serum-mediated inhibition and in enhanced efficiency of gene transfer. Sucrose density gradient sedimentation analysis indicated that the VSV-G associates physically with the DNA-lipid complex to produce a VSV-G liposome. The ability to incorporate surrogate viral or cellular envelope components such as VSV-G into liposomes may allow more-efficient and possibly targeted gene delivery by lipofection, both in vitro and in vivo.

scholarly journals Recombinant Retroviruses Pseudotyped With the Vesicular Stomatitis Virus G Glycoprotein Mediate Both Stable Gene Transfer and Pseudotransduction in Human Peripheral Blood Lymphocytes

Blood ◽  
1997 ◽  
Vol 90 (3) ◽  
pp. 952-957 ◽  
Author(s):  
H.F. Gallardo ◽  
C. Tan ◽  
D. Ory ◽  
M. Sadelain
Keyword(s):  
Gene Transfer ◽  
Vesicular Stomatitis Virus ◽  
Peripheral Blood ◽  
Cell Function ◽  
Human Peripheral Blood ◽  
Peripheral Blood Lymphocytes ◽  
Stable Gene ◽  
Cell Therapies ◽  
Blood Lymphocytes ◽  
Vesicular Stomatitis

Abstract It is essential for the study of T-cell function and the improvement of adoptive cell therapies to efficiently generate large populations of human primary T cells that reliably express foreign genes. This goal is achieved by using recombinant retroviruses pseudotyped with either the gibbon ape leukemia virus (GaLV) envelope or the vesicular stomatitis virus G (VSV-G) glycoprotein. We show here that both retroviral particles mediate stable gene transfer in CD4+ and in CD8+ peripheral blood lymphocytes cultured under optimized conditions. However, VSV-G–pseudotyped virions may cause transduction artifacts that must be carefully excluded. The VSV-G virions require 10- to 100-fold higher concentrations of infectious particles to achieve levels of gene transfer comparable to GaLV-virions. Nonetheless, the physical stability of VSV-G–coated particles enables the concentration of viral stocks to 109 infectious particles per milliliter or more.

scholarly journals Use of DNA-mediated gene transfer to analyze the role of H-2Ld in controlling the specificity of anti-vesicular stomatitis virus cytotoxic T cells.

1983 ◽  
Vol 157 (4) ◽  
pp. 1261-1272 ◽  
Author(s):  
J Forman ◽  
R S Goodenow ◽  
L Hood ◽  
R Ciavarra
Keyword(s):  
T Cells ◽  
Gene Transfer ◽  
Cell Line ◽  
Vesicular Stomatitis Virus ◽  
Cytotoxic T Cells ◽  
Control Cell ◽  
Target Cells ◽  
Effector Cells ◽  
Vesicular Stomatitis ◽  
Simplex Virus

Mouse thymidine kinase (tk-) C3H L (H-2k) cells transformed by the technique of DNA-mediated gene transfer with the herpes simplex virus tk gene together with the BALB/c H-2Ld gene express H-2Ld molecules indistinguishable from their counterparts on spleen cells. An established cloned cell line (8-5) was used to assess the function of the H-2Ld antigen in determining the specificity of alloreactive as well as anti-vesicular stomatitis virus (VSV) cytotoxic T cells (CTL). Both anti-H-2d and anti-H-2Ld CTL displayed a cytotoxic effect against 8-5 cells but not a control cell line transformed with the tk gene only (tk+ cells). Further evidence that 8-5 cells express H-2Ld was provided by the finding that monoclonal anti-H-2Ld but not H-2Dd antibodies blocked target cell lysis by the effector cells. Both BALB/c (H-2d) and DBA/2 (H-2d) animals generated anti-VSV CTL that lysed infected 8-5 but not tk+ cells. To further establish that H-2Ld controlled the specificity of the effector cells, a monoclonal antibody directed against H-2Ld was shown to inhibit lysis of infected 8-5 target cells. To determine whether other H-2d-encoded gene products could serve as restricting antigens for anti-VSV CTL in BALB/c animals, unlabeled VSV infected 8-5 cells were tested for their ability to block lysis of 51chromium-labeled P815 (H-2d)-infected target cells. The 8-5-VSV inhibitor cells inhibited lysis to a slightly lesser extent than unlabeled P815-VSV cells, indicating that H-2Ld plays a major if not exclusive role in restricting anti-VSV CTL in H-2d animals.

One-Day Ex Vivo Culture Allows Effective Gene Transfer Into Human Nonobese Diabetic/Severe Combined Immune-Deficient Repopulating Cells Using High-Titer Vesicular Stomatitis Virus G Protein Pseudotyped Retrovirus

Blood ◽  
1999 ◽  
Vol 93 (7) ◽  
pp. 2217-2224 ◽  
Author(s):  
Vivienne I. Rebel ◽  
Mayumi Tanaka ◽  
Jeng-Shin Lee ◽  
Sheila Hartnett ◽  
Michael Pulsipher ◽  
...  
Keyword(s):  
Stem Cell ◽  
Gene Transfer ◽  
G Protein ◽  
Vesicular Stomatitis Virus ◽  
Ex Vivo ◽  
High Titer ◽  
Hematopoietic Stem ◽  
Nonobese Diabetic ◽  
Vesicular Stomatitis ◽  
Ex Vivo Culture

Retrovirus-mediated gene transfer into long-lived human pluripotent hematopoietic stem cells (HSCs) is a widely sought but elusive goal. A major problem is the quiescent nature of most HSCs, with the perceived requirement for ex vivo prestimulation in cytokines to induce stem cell cycling and allow stable gene integration. However, ex vivo culture may impair stem cell function, and could explain the disappointing clinical results in many current gene transfer trials. To address this possibility, we examined the ex vivo survival of nonobese diabetic/severe combined immune-deficient (NOD/SCID) repopulating cells (SRCs) over 3 days. After 1 day of culture, the SRC number and proliferation declined twofold, and was further reduced by day 3; self-renewal was only detectable in noncultured cells. To determine if the period of ex vivo culture could be shortened, we used a vesicular stomatitis virus G protein (VSV-G) pseudotyped retrovirus vector that was concentrated to high titer. The results showed that gene transfer rates were similar without or with 48 hours prestimulation. Thus, the use of high-titer VSV-G pseudotyped retrovirus may minimize the loss of HSCs during culture, because efficient gene transfer can be obtained without the need for extended ex vivo culture.

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