Development of the Molecular Biology of HIV-Seek: A Novel Approach to Characterizing Latently Infected CD4+ T Cells
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2022
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Although antiretroviral therapy (ART) is effective at managing HIV infection and extending the lives of people living with HIV (PLWH), it is limited by the inability to target the HIV viral reservoir, specifically in latently infected CD4+ T cells. Strategies for targeting the viral reservoir, including “shock and kill” and “block and lock,” while still in development, require more supportive clinical data to legitimize this approach. These strategies may benefit from an accurate characterization of latently infected cells. However, current techniques for characterization are limited due to the scarcity of latency and the stipulation to activate latently infected cells. Cell reprogramming in the activated state of CD4+ T cells obscures the biology of the cell in its latent state. Therefore, we introduce HIV-Seek, a novel approach to characterize the pre-activated transcriptome of latently infected resting memory CD4+ T cells. This characterization will reveal more information on an infected cell in its latent state and may then enable the identification of a biomarker unique to latently infected cells. In turn, a successful biomarker candidate could lead to immunological or pharmacological control of the viral reservoir. HIV-Seek utilizes a variety of technologies including microfluidics and single-cell transcriptomics to isolate and analyze the pre-activated transcriptome of latently infected cells. In this thesis, we establish bead-based molecular techniques that demonstrate the use of oligo-dT-conjugated beads to facilitate reactions involved in HIV-Seek.
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Molecular biology, Virology, Immunology, CD4+ T cells, HIV, HIV latency, HIV-Seek
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100 pages
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