Date of Graduation

Spring 2019

Degree

Master of Science in Cell and Molecular Biology

Department

Biomedical Sciences

Committee Chair

Amy Hulme

Keywords

HIV-1, viral replication, uncoating, TRIM-CypA, microglia

Subject Categories

Cell Biology | Molecular Biology | Virology

Abstract

Uncoating is a poorly understood yet required step of HIV-1 replication that is defined as the disassembly of the viral capsid structure. The goal of this project is to characterize uncoating in C20 microglial cells. These cells are a natural target of HIV-1 that are infected to establish latent viral reservoirs and HIV-associated neurological disorders. A stable C20 cell line that expresses TRIM-CypA was established to study the kinetics of uncoating with the CsA washout assay. The expression of TRIM-CypA was confirmed by western blot and the functionality of the protein was confirmed by a viral infectivity assay. Using this cell line, uncoating was found to have an average half-life of 63.3 min. Viral fusion assays determined the kinetics of viral fusion in C20 cells compared to uncoating. The average half-life of viral fusion was found to be 40.9 min. To characterize the effect of CsA on HIV infectivity in C20 cells a viral infectivity assay was performed in the presence and absence of CsA. It was discovered that HIV infectivity was slightly lower in the presence of CsA. The study of uncoating kinetics in microglial cells will give more insight on how HIV establishes infection in brain cells which contributes to a larger goal of developing better therapies, a vaccine, and ultimately, a cure to HIV.

Copyright

© Melanie Anne Taylor

Open Access

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