PhD, Virology, Faculty of Health Sciences, University of the Witwatersrand / HIV-associated cardiomyopathy is a multifactorial disease with a broad spectrum of aetiologies that arise due to chronic immunosupression during HIV infection. The intricate relationship between HIV infection and cardiac co-morbidity was investigated with the aid of HIV-neutralizing aptamers. These synthetic nucleic acid ligands with antibody-like properties are molecular tools with multifunctional applications ranging from drug discovery to diagnostics and therapeutics. The advent of the HIV/AIDS pandemic has naturally married the field of HIV therapy and diagnostics with that of aptamer technology. By employing a HIV-1 neutralizing aptamer, named UCLA1, raised against the viral surface envelope glycoprotein 120, I dissected some of the pathways leading to cardiomyocyte apoptosis in a cell culture system. In chapter one I investigated the potential cytotoxic effects of UCLA1 by comparing it against a panel of 17 antiretrovirals (ARVs) in clinical use with the goal of establishing a safety portfolio geared towards its use as a therapeutic agent. Using cultured human cardiomyocytes and primary peripheral blood mononuclear cells (PBMCs), I selected some of the major biological markers of ARV-induced cytotoxicity and found no measurable deleterious effect, especially when compared to other ARVs used in the same study. In chapter two, the permissiveness of cardiomyocytes to HIV infection as well as the relationship between virus-host interaction and caspase-mediated apoptosis were investigated. Non-productive, receptor and tropism-independent infection was observed, which was arrested after the reverse transcription stage. However, interaction between the virus gp120 and the host’s CXCR-4 chemokine receptor preferentially activated caspase-9 triggering robust mitochondria-mediated apoptosis. A shift from mitochondrial-initiated, caspase-9 mediated to Fas-ligand initiated, caspase-8 mediated was observed when CM were co-cultured with HIV-infected MDM. UCLA1 protected against caspase-9 mediated
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apoptosis but not caspase-8 mediated. Finally in chapter three I provided answers for the shift in caspase activation by showing that supraphysiological levels of IL-1β and IL-6 during HIV infection of MDM augment the effects of tumor necrosis factor (TNF). These observation provide new insight into the complex pathophysiology of HIVCM and highlight the potential of UCLA1 as a novel therapeutic agent to fight HIV and some of its associated diseases.
| Identifer | oai:union.ndltd.org:netd.ac.za/oai:union.ndltd.org:wits/oai:wiredspace.wits.ac.za:10539/8996 |
| Date | 02 February 2011 |
| Creators | Rangel Lopes de Campos, Walter |
| Source Sets | South African National ETD Portal |
| Language | English |
| Detected Language | English |
| Type | Thesis |
| Format | application/pdf |
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