Date of Graduation

Summer 2022

Degree

Master of Science in Cell and Molecular Biology

Department

Biomedical Sciences

Committee Chair

Amy Hulme

Abstract

Microglial cells are immune cells that protect the central nervous system and are subject to HIV- 1 infection. HIV-1 infection of these cells can lead to HIV Associated Neurocognitive Disorders (HAND). Since HIV-1 only codes for 15 different proteins, the virus also uses cellular factors, like Cyclophilin A (CypA), to assist in viral replication. The cell also has antiretroviral restriction factors, like tripartite motif-containing protein 5 (TRIM5α), to protect against infection. The goal of this project is to investigate the role of CypA in the early steps of HIV-1 replication and TRIM5α restriction in microglial cells. I hypothesize that CypA facilitates HIV-1 infection and assists in normal capsid uncoating kinetics. I also predict that blocking CypA interaction will allow for TRIM5α restriction to decrease HIV-1 infection. To study how the interaction between CypA and the capsid affects uncoating, the in situ uncoating assay was performed in the presence and absence of the drug Cyclosporin A which blocks this interaction. Lower amounts of CA were found in conditions with CypA binding compared to the absence of CypA. This result suggests that CypA assists with uncoating in CHME3 cells. To research the interplay between CypA and TRIM5α in microglial cells, TRIM5α expression was knocked down using siRNAs and CsA treatment was used to block CypA binding to the capsid. A decrease in TRIM5α expression resulted in an increase of HIV-1 infectivity. This effect was greater when CypA interactions were inhibited. This data indicates that TRIM5α restriction occurs in CHME3 microglial cells when CypA is inhibited. It is likely that CypA plays more than one role in HIV-1 infectivity and replication. Here, I have characterized the effect of CypA on HIV-1 uncoating and the connection between CypA and TRIM5α in microglial cells.

Keywords

HIV-1, uncoating, cyclophilin A, TRIM5α, microglial cells

Subject Categories

Medicine and Health Sciences | Virus Diseases

Copyright

© Emma LK Wise

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