Recombinant lymphocytic choriomeningitis virus expressing vesicular stomatitis virus glycoprotein.

Abstract

A recombinant S segment RNA (Sr) of the prototypic arenavirus lymphocytic choriomeningitis virus (LCMV) where the glycoprotein of vesicular stomatitis virus (VSVG) was substituted for the glycoprotein of LCMV (LCMV-GP) was produced intracellularly from cDNA under the control of a polymerase I promoter. Coexpression of the LCMV proteins NP and L allowed expression of VSVG from Sr. Infection of transfected cells with WT LCMV (LCMVwt) resulted in reassortment of the L segment of LCMVwt with the Sr at low frequency. Isolation of recombinant LCMV (rLCMV) expressing VSVG (rLCMV/VSVG) was achieved by selection against LCMVwt by using a cell line deficient in the cellular protease S1P. This approach was based on the finding that processing of LCMV-GP by S1P was required for virus infectivity. Characterization of protein and RNA expression of rLCMV/VSVG in infected cells confirmed the expected virus genome organization. rLCMV/VSVG caused syncytium formation in cultured cells and grew to approximately 100-fold lower titers than WT virus but, like the parent virus, it persisted in neonatally infected mice without clinical signs of disease.

• Authors: Daniel D Pinschewer, Mar Perez, Ana B Sanchez, Juan Carlos de la Torre
• Journal: Proceedings of the National Academy of Sciences of the United States of America (Proc Natl Acad Sci U S A) Vol. 100 Issue 13 Pg. 7895-900 (Jun 24 2003) ISSN: 0027-8424 [Print] United States
• PMID: 12808132 (Publication Type: Journal Article)

Chemical References

• G protein, vesicular stomatitis virus
• Membrane Glycoproteins
• Viral Envelope Proteins
• RNA

Topics

• Animals
• Blotting, Northern
• Blotting, Western
• CHO Cells
• Chlorocebus aethiops
• Cricetinae
• Genetic Engineering
• Genetic Techniques
• Lymphocytic choriomeningitis virus (genetics, metabolism)
• Membrane Glycoproteins (biosynthesis, genetics)
• Mice
• Mice, Inbred BALB C
• Microscopy, Fluorescence
• Plasmids (metabolism)
• RNA (metabolism)
• Reverse Transcriptase Polymerase Chain Reaction
• Time Factors
• Transfection
• Vero Cells
• Viral Envelope Proteins (biosynthesis, genetics)