Vesicular stomatitis virus G glycoprotein pseudotyped retroviral vectors: concentration to very high titer and efficient gene transfer into mammalian and nonmammalian cells.

Abstract

The restricted host-cell range and low titer of retroviral vectors limit their use for stable gene transfer in eukaryotic cells. To overcome these limitations, we have produced murine leukemia virus-derived vectors in which the retroviral envelope glycoprotein has been completely replaced by the G glycoprotein ofvesicular While retroviral infection usually requires interaction between the viral envelope protein and specific cell surface receptor proteins, VSV-G interacts with a phospholipid component of the cell membrane to mediate viral entry by membrane fusion (6). Since viral entry seems not to be dependent on the presence of specific protein receptors, VSV has an extremely broad host-cell range (7). In addition, VSV can be concentrated by ultracentrifugation without loss of infectivity. We hypothesized that substitution of VSV-G for the MoMLV envelope protein might confer upon the pseudotyped particle the desirable properties of increased host-cell range and structural stability after ultracentrifugation. We report here the construction of a retroviral vector encapsidated in the envelope protein of VSV that has an extended host-cell range and can be concentrated to titers >109 colony-forming units (cfu)/ml with minimal loss of infectivity. As

Truncated vesicular stomatitis virus G protein improves baculovirus transduction efficiency in vitro and in vivo

“…11 In insect cells, expression of the VSV-G led to cell fusion during virus production. Preparation of the VSV-GED-pseudotyped virus, however, did not show this adverse effect.…”

“…6 One of the most widely used pseudotyping tools is G glycoprotein of the vesicular stomatitis virus (VSV-G). [7][8][9][10] VSV-G is routinely used to enhance the target range and transduction efficiency of retroviruses 11 by providing wider tropism as well as improved viral stability and augmented resistance to the complement inactivation. 12 Several reports of VSV-G pseudotyped baculoviruses provide evidence that VSV-G is able to enhance transduction efficiency of baculovirus in vertebrate cells in vitro and in vivo.…”

In vivo transduction of mouse epidermis with recombinant retroviral vectors: implications for cutaneous gene therapy

“…17,18 On day 3 after abrasion 50 l of infectious VSV-G pseudotyped RRV-LZRN (1 × 10 9 c.f.u./ml) or recombinant Ad-LZ (5 × 10 10 p.f.u./ml) encoding the E. coli LacZ reporter gene were injected in the plane between the scab and the re-epithelializing surface. The latter was used as a control to assess accessibility of keratinocytes as well as turnover rate of the transgene and the transduced cells.…”

“…)/ml as determined on NIH 3T3 cells. 18 Ad-LZ is a replicationdefective EI − , E3 − deletion mutant expressing LacZ under the control of cytomegalovirus promoter. The vector was propagated and purified as previously described.…”

MuLV-based vectors pseudotyped with truncated HIV glycoproteins mediate specific gene transfer in CD4+ peripheral blood lymphocytes

“…[3][4][5][6][7][8] However, targeted delivery of a murine-based retroviral vector to a specific subpopulation of human cells has yet to be truly achieved. Initial attempts to change viral tropism were done through the use of pseudotyped viruses; [9][10][11] MuLV is able to incorporate vesicular stomatitis virus G glycoprotein (VSV-G), influenza virus hemagglutinin (HA), or heterologous retroviral glycoproteins, although this expands viral host range rather than restrict cell tropism. Even though several groups have generated chimeric Envligand fusion proteins designed to bind on to transferrin receptor, galactose receptor or the high density lipoprotein receptor, none of these fusion proteins were successfully incorporated into infectious viral particles.…”