Pseudotype Formation of Moloney Murine Leukemia Virus with Sendai Virus Glycoprotein F

Abstract

Mixed infection of cells with both Moloney murine leukemia virus (MoMLV) and related or heterologous viruses produces progeny pseudotype virions bearing the MoMLV genome encapsulated by the envelope of the other virus. In this study, pseudotype formation between MoMLV and the prototype parainfluenza virus Sendai virus (SV) was investigated. We report for the first time that SV infection of MoMLV producer cells results in the formation of MoMLV(SV) pseudotypes, which display a largely extended host range compared to that of MoMLV particles. This could be associated with SV hemagglutinin-neuraminidase (SV-HN) glycoprotein incorporation into MoMLV envelopes. In contrast, solitary incorporation of the other SV glycoprotein, SV fusion protein (SV-F), resulted in a distinct and narrow extension of the MoMLV host range to asialoglycoprotein receptor (ASGP-R)-positive cells (e.g., cultured human hepatoma cells). Since stably ASGP-R cDNA-transfected MDCK cells, but not parental ASGP-R-negative MDCK cells, were found to be transduced by MoMLV(SV-F) pseudotypes and transduction of ASGP-R-expressing cells was found to be inhibited by ASGP-R antiserum, a direct proof for the ASGP-R-restricted tropism of MoMLV(SV-F) pseudotypes was provided. Cultivation of ASGP-R-positive HepG2 hepatoma cells on Transwell-COL membranes led to a significant enhancement of MoMLV(SV-F) titers in subsequent flowthrough transduction experiments, thereby suggesting the importance of ASGP-R accessibility at the basolateral domain for MoMLV(SV-F) pseudotype transduction. The availability of such ASGP-R-restricted MoMLV(SV-F)-pseudotyped vectors opens up new perspectives for future liver-restricted therapeutic gene transfer applications.

Viral vectors: from virology to transgene expression

“…, 2001), arenaviruses (Miletic et al. , 1999), hepadnaviridae (Sung and Lai, 2002), paramyxoviridae (Spiegel et al. , 1998; Kobinger et al.…”

Lentiviral Vectors Pseudotyped with Envelope Glycoproteins Derived from Human Parainfluenza Virus Type 3

“…Another approach might be to genetically modify the envelope proteins to increase their level of incorporation. This approach has been successfully used to increase the level of incorporation of heterologous envelope proteins by truncation of their cytoplasmic domains, fusion with the cytoplasmic domains of retroviral or lentiviral envelope proteins, or through the use of more sophisticated modifications designed to relieve steric hindrance or alter envelope protein folding (26)(27)(28)(29).…”

Sendai virus vectors as an emerging negative‐strand RNA viral vector system

“…Tissue‐specific infection of these particles could be demonstrated by an interaction of F with the liver‐specific asialoglycoprotein receptor (ASGP‐R) followed by a productive infection of liver‐derived cells 6. This interaction of the F protein with the ASGP‐R could be further demonstrated for SeV wild‐type viruses 49, for recombinant HN‐devoid virus‐like particles 50, as well as for a F‐pseudotyped retroviral 51 and lentiviral vector 52. However, the titers reported for these pseudotyped vectors to date have been very low; as a consequence, a detailed characterization of the F‐mediated tissue restriction in vivo can only be performed after substantially increasing the titers in these systems or when SeVV‐ΔHN particles become available, which have not been reported yet.…”