Development of a next generation quantitative viral outgrowth assay QVOA for the standardized measurement of the HIV latent reservoir

Project Summary Abstract Human immunodeficiency virus type HIV is a retrovirus that infects CD T cells of the immune system If left untreated HIV 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 in infected individuals Despite the success of this therapy at suppressing HIV replication to clinically undetectable levels antiretroviral therapy is not curative This is due to the persistence of HIV in a silent or latent state within a subset of CD T cells known as resting memory CD 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 infected individuals latently infected CD T cells are found at extremely low frequencies per million resting memory CD T cells However this population of latently infected cells is very stable demanding that HIV infected individuals remain on antiretroviral therapy indefinitely Therefore this population of latently infected CD T cells is the main barrier to curing HIV infection Developing strategies to eliminate latently infected cells is a major focus of the NIH NIAID and the HIV 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 The currently accepted gold standard assay for measuring latent HIV is the quantitative viral outgrowth assay developed by the Siliciano laboratory in the mid s Due to the complexity and high resource requirements of the QVOA this assay can only be performed in a small number of research laboratories Accelevir Diagnostics LLC is developing a new quantitative viral outgrowth assay with improved precision reproducibility and scalability Broadly this proposal aims to optimize assay conditions and perform key assay validation studies including laboratory automation studies The goal of this proposal is to develop an optimized commercial prototype with accompanying standard operating procedures and set the stage for rapid analytical validation and market entry !Project Narrative For HIV infected individuals on suppressive antiretroviral therapy latent HIV infection of resting memory CD 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 scalable assay to measure the number of latently infected cells in patients by Accelevir Diagnostics for use in monitoring the efficacy of HIV cure therapies