Human immunodeficiency virus type-1 (HIV-1) is a retrovirus that infects CD4+ T cells of the immune system. If left untreated, HIV-1 infected individuals will progress to AIDS and may ultimately die as a result. Combination antiretroviral therapy is extremely effective at stopping the replication of HIV-1 in infected individuals. Despite the success of this therapy at suppressing HIV-1 replication to clinically undetectable levels, antiretroviral therapy is not curative. This is due to the persistence of HIV-1 in a silent, or latent, state within a subset of CD4+ T cells known as resting memory CD4+ T cells. In this latent state, these infected cells are not targeted by antiretroviral drugs and cannot be eliminated by the immune system. In HIV-1 infected individuals, latently infected CD4+ T cells are found at extremely low frequencies (~1 per million resting memory CD4+ T cells). However, this population of latently infected cells is very stable, demanding that HIV-1 infected individuals remain on antiretroviral therapy indefinitely. Therefore, this population of latently infected CD4+ T cells is the main barrier to curing HIV-1 infection. Developing strategies to eliminate latently infected cells is a major focus of the NIH, NIAID, and the HIV-1 research field. To demonstrate the efficacy of therapeutics targeting the latent reservoir, we must be able to measure the frequency of latently infected cells using rapid and accurate assays that can be scaled for widespread clinical use. However, such assays are not currently available. Accelevir Diagnostics, LLC is therefore developing a new molecular assay to accurately measure the size of the latent reservoir by quantifying the number of intact, replication competent proviruses present in CD4+ T cells from HIV-1 infected patients. Broadly, this proposal aims to optimize sample preparation and assay conditions as well as plan and collect samples for analytical validation studies. The goal of this proposal is to develop an optimized commercial prototype of our molecular assay for the HIV-1 latent reservoir, with accompanying standard operating procedures, and set the stage for rapid analytical validation and market entry.
For HIV-1 infected individuals on suppressive antiretroviral therapy, latent HIV-1 infection of resting memory CD4+ T cells is the major barrier to a cure. Elimination of these latently infected cells by has been proposed as a strategy to cure infected individuals. This proposal details the development and optimization of a new molecular assay to measure the number of latently infected cells in patients by Accelevir Diagnostics, for use in monitoring the efficacy of HIV-1 cure therapies.
