Related Experiment Videos
Expression of biologically active envelope glycoprotein from the acutely pathogenic simian immunodeficiency virus
S A González1, J L Affranchino, A Burny
1Department of Molecular Biology, University of Brussels (ULB), Rhode-St.-Genèse, Belgium.
Abstract:
The full-length envelope (env) gene from the most acutely pathogenic primate lentivirus described so far, the simian immunodeficiency virus SIVsmmPBj14 was expressed by a recombinant vaccinia virus vector (vv-env4) and was completely characterized as a previous step for its use as an immunogen in vaccination trials. Radioimmunoprecipitation and Western blot experiments indicated that SIVsmmPBj gp160 precursor was processed into gp120 and gp41 subunits, and that gp120 was released into the medium. Flow cytometry analysis showed that recombinant SIVsmmPBj was transported to and expressed on the surface of vvenv4-infected cells. Biochemical analysis of virus-like particles produced by coinfection of cells with recombinant vaccinia viruses expressing SIVsmmPBj Env (vv-env4) and Gag (vv-wtgag) proteins revealed that the Env glycoprotein was incorporated into core-like particles. Furthermore, cells expressing SIVsmmPBj env gene products were found to undergo fusion with the same CD4+ cell lines in which the whole provirus has been shown to form syncytia.
Insights
Researchers expressed the simian immunodeficiency virus (SIV) envelope gene using a vaccinia virus vector. This characterization is a crucial step for developing SIV vaccines and understanding lentivirus pathogenesis.
Area of Science:
- Virology
- Immunology
- Molecular Biology
Background:
- The simian immunodeficiency virus (SIV) SIVsmmPBj14 is a highly pathogenic primate lentivirus.
- Understanding the SIV envelope (env) gene is critical for developing effective vaccines and antiviral therapies.
Purpose of the Study:
- To express and characterize the full-length SIVsmmPBj14 envelope (env) gene using a recombinant vaccinia virus vector (vv-env4).
- To evaluate the potential of the expressed SIV env gene products as immunogens for vaccination trials.
Main Methods:
- Recombinant vaccinia virus vector (vv-env4) was used to express the SIVsmmPBj14 env gene.
- Radioimmunoprecipitation and Western blot assays were performed to analyze protein processing.
- Flow cytometry was used to assess surface expression of the SIV Env protein.
- Biochemical analysis of virus-like particles (VLPs) was conducted.
Main Results:
- The SIVsmmPBj14 gp160 precursor was processed into gp120 and gp41 subunits, with gp120 secreted into the medium.
- The recombinant SIV Env protein was expressed on the surface of infected cells.
- Env glycoproteins were incorporated into SIV-like particles.
- Cells expressing SIV Env induced cell fusion.
Conclusions:
- The characterization of the SIVsmmPBj14 env gene expression is complete.
- The expressed SIV Env protein is functional and can be incorporated into virus-like particles.
- These findings support the use of the SIV env gene as an immunogen in future vaccine development.