Melanie Alvarado, Master of Science candidate in biological sciences, will be defending her thesis, “Defining molecular pathways in the transcriptional reactivation of latently infected HIV-1 T-Cell lines” on Thursday, May 4, at 10 a.m. in Room 307 of the UAA/APU Consortium Library.
As of 2012, in the United States, there have been reports of 1.2 million people living with human immunodeficiency virus (HIV). Of these individuals, roughly 161,200 are unaware they are infected with HIV, per the CDC. HIV was once a death sentence, but is now manageable with the assistance of Antiretroviral Therapy (ART). Unfortunately, this treatment combination does not eradicate HIV due to the quiescent reservoir lingering in the patient. If a treatment is interrupted or stopped, viral loads will again increase, eventually leading to AIDS. Accompanying health concerns from negative side effects, there is also an economic burden for patients.
Novel approaches are required to establish which molecular factors are involved in maintaining HIV latency in order to contribute to a functional cure for HIV/AIDS. Preliminary research has identified molecular pathways in cells containing latent HIV. Targeting these pathways with synthetic viral RNA should expose the latent HIV integrated in the host chromatin, allowing it to be eliminated by antiviral drug therapy. This “shock and kill” strategy is currently in clinical trials. However, the consequences of exposing latent HIV by these strategies are unknown. Viruses activate innate immune (interferon) pathways in the cell.
Using cultured HIV GFP reporter T-cells in the lab, the experiments described in this thesis will examine how reactivation of latent HIV can be triggered by interferon pathways. Mapping which interferon pathway is activated upon viral RNA poly-I:C stimulation can contribute to determining which factors are involved in HIV reactivation. I hypothesize that poly-I:C will induce activation of the innate immune response via IRF3 and NFkB pathways, as well as reactivate the latent HIV provirus reservoir observable with green fluorescent protein (GFP). Results obtained demonstrated poly-I:C treatments strongly induce HIV reactivation in latently infected T-cell lines. Transcription factor IRF3 was found to be essential for HIV reactivation with poly-I:C treatments and strongly suggests NFkB is also required to initiate activation. Transcription factor IRF7 was found to not be required for HIV reactivation. These findings can be used to improve current treatments for HIV-positive patients, reducing negative side effects such as susceptibility to secondary co-morbidities.