Traditional vaccine methods have long been employed to control widespread infectious diseases, but so far, all commercially available vaccine strategies have been inadequate in efforts to develop an effective therapeutic HIV vaccine. However, recent advancements in immunological research have lead to the generation of novel vaccine strategies, one of which is the recombinant virus vaccine, a method of particular interest that has shown promise in the clearance of HIV infection within HIV-positive patients who have retained immunocompetence. This study examined the stability of the expression of HIV-1 genes, gag and env, through a recombinant virus vector, a recombinant vesicular stomatitis virus (VSV), a temperature-sensitive mutant genetically modified to contain the select HIV-1 genes (VSVInd(GML)HIV-1gag-env). For SDS-PAGE, incubation was at 31 °C and 37 °C, the permissible and semi-permissible growth conditions respectively for VSVInd(GML) temperature-sensitive mutants. Western blot analysis was used to quantitate levels of protein expression of full VSV proteins, Gag and Env, using a primary rabbit antibody of anti-VSV anti-serum and a secondary anti-IgG from rabbit, a primary antibody of anti-p24 anti-serum and a secondary anti-IgG from rabbit, and a primary goat antibody of anti-gp120, and a secondary anti-IgG from goat. Results indicated that the VSVInd(GML) vector system allowed for high levels of expression of HIV-1 gag and env genes. It is known that the expression of these genes induces the production of major neutralizing antibodies and the stimulation of cytotoxic T lymphocytes, revealing the potential to use a genetically modified recombinant VSV as a universal vector for the development of recombinant virus vaccines. Specifically, the VSVInd(GML) mutant vector is thus an attractive candidate for the viral vector of a therapeutic HIV vaccine system.
Baek, Justine, "Expression of HIV-1 Gag and Env Genes Using the Vesicular Stomatitis Virus Vector System" (2015). 2015 Undergraduate Awards. 19.