Description
Ever since the start of the epidemic of Human Immunodeficiency Virus (HIV), the causative agent of AIDS, there has been over 35 million deaths. Today, about 37 million people live with HIV-1 and about 5,000 new patients become infected daily. This virus affects people with various ethnic backgrounds, ages, gender, and economic classes, and about 95% of the patients live in developing countries. HIV is one of the most diverse, universal, and prevalent pathogens any researcher or medical professional has ever faced, so the need for more affordable, more efficient, and less painful antivirals and novel treatments for HIV, must be a priority within medical research today. Within most viral life cycles, interactions between the host cell and the virus occur. These viral-host interactions take place within the classical secretory pathway of an eukaryotic cell. RNA viruses, like HIV, utilize the vesicles of the secretory pathway to help aid in the maturation and transport of their glycoproteins to the cell surface in order to infect other cells within the immune system. As the envelope polyprotein of HIV travels through the secretory pathway, it will be cleaved into its viral, active form. The infectious viral progeny will be transported to the cell surface to continue the spreading of the virus. Inhibiting or disrupting the cleavage event of the HIV Envelope protein (Env) represents an attractive target for drug development. Within medical research, cell based assays are key components when streamlining the drug development process. They are analytical tools used to study different mechanisms of cellular functions. These advantages have led companies, universities, and laboratories to save both time and money. Outlined in this thesis is the adaptations engineered into a previous established cell-based assay. This cell-based assay monitors the transport and cleavage events of the HIV Env protein while traveling through the classical secretory pathway. Larger segments of the Env protein were engineered into this assay to provide insight into which domains directly affect these events and will validate the Env protein as a novel target for future discoveries of antivirals against HIV-1.
