Date of Award
Wayne L. Gray
Described as one of the world’s worst pandemics, HIV (Human Immunodeficiency Virus) infects millions of people each year and is the cause for AIDS (Acquired Immunodeficiency Syndrome). Despite the development of vaccines for numerous infectious diseases such as polio, small pox, and influenza, a vaccine for HIV remains elusive due to the virus’s high mutation rate and ability to evade the immune system. HIV causes depletion of CD4+ lymphocytes, resulting in a weakened immune system. However, the development of a plasmid-based DNA vaccine approach may help revolutionize vaccine development for HIV due to its ability to confer cellular and humoral immunity through T-cells and antibodies, respectively. The already constructed pVAX1 plasmid’s small size and multiple cloning sites make it an effective vector for the development of a plasmid-based DNA vaccine for HIV. In this experiment, a gene sequence for SIV (Simian Immunodeficiency Virus) proteins inserted into the pVAX1 plasmid was transfected into an African green monkey kidney cell line (Vero cells) for expression of SIV proteins.
HIV and AIDS research aims to contribute to the development of prevention and treatment strategies for this disease. The research in this thesis focuses on characterization of a plasmid-based DNA vaccine expressing rev, tat, and nef (Retanef) proteins for SIV using restriction endonuclease analysis, PCR, immunofluorescence, and western blotting. The specific aims for this thesis include: evaluating the presence of the Retanef gene sequence inside pVAX-RTN, and analyzing the expression of the Retanef protein in vitro. This study showed that transfection of pVAX-RTN into Vero cells resulted in successful expression of the Retanef protein. Detection of the Retanef protein by SIV positive monkey sera also showed positive results, further proving that Retanef is expressed inside Vero cells via the pVAX-RTN plasmid.
The results acquired in this experiment will contribute towards HIV vaccine research by using SIV as the experimental model for HIV. The results acquired in this research supports the use of early regulatory proteins in SIV vaccines and will be contribute towards successfully creating an effective HIV vaccine in the future.
Sanipara, Priya, "Characterization of a Plasmid-Based DNA Vaccine for Simian Immunodeficiency Virus" (2020). Honors Theses. 1502.
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