Standardizable Whole Blood Culture for Evaluating HIV Eradication Strategies

DESCRIPTION provided by applicant The impetus for new andquot curativeandquot approaches for HIV AIDS lacks adequate clinical tools to monitor patient responses to therapeutics targeting the persistent viral reservoir We therefore propose to develop a standardizable integrated clinically relevant whole blood culture assay to evaluate HIV eradication therapeutics ex vivo by directly measuring responses of primary peripheral blood cells under internally referenced patient specific matched control and therapeutic conditions The proposed system will quantify single cycle virus production from infected primary blood cells comprising the entire peripheral blood immunological compartment The relative difference in virus production under latency activating conditions versus unstimulated matched cultures represents the target replication competent provirus residing in latently infected primary blood cells The culture assay will support ex vivo testing of latency activating cytolytic immunomodulatory and other andquot curativeandquot therapeutic approaches required by the FDA for therapeutic success This standardizable readily accessible whole blood culture assay for testing HIV eradication strategies will provide an enabling next generation research clinic and patient management platform The Jericho team proposes to validate the feasibility of developing such a platform here via this ambitious Phase I SBIR project expanded scope and budget The envisioned integrated blood collection system for HIV infected whole blood culture is a closed device requiring less than mL of donor blood per collection tube directly obtained by a phlebotomist using a standard blood draw After up to hours of incubation at C in a proprietary media viable cells soluble factors and virions can be co collected and measured from these primary blood cultures to provide correlative biomarkers The proposed product will allow for comparison of different test conditions e g latency activators and therapeutics e g suppressive drugs cytokines across multiple internally referenced culture tubes specific to the infection status and immunological response of the individual We propose to demonstrate the feasibility of standardizing and developing this platform to evaluate endpoints for HIV eradication strategies by characterizing virus production cellular markers and soluble factors from HIV infected donor whole blood sample collections We will first quantify and characterize reproducibility of our stimulated virus production culture system within a coefficient of variance between replicate matched collection tubes We will then demonstrate the potential utility of the characterized platform using two selected compounds known to stimulate latently infected cells Preclinical therapeutic optimization translates directly to clinical monitoring and patient management having identified relevant conditions and associated biomarkers using the proposed whole blood culture assay platform As HIV is now moving from a manageable chronic disease to a andquot cure andquot novel diagnostics will be increasingly required to ensure that the best treatments are used to optimize efficacy and to reduce patient risks PUBLIC HEALTH RELEVANCE This Phase I SBIR project is designed to prove the feasibility of developing a comprehensive whole blood culture platform useful for clinically testing therapeutics intended for eradication of HIV infection This work represents a key step toward producing a next generation research and clinical platform that will enable the optimization of therapeutic treatments for individuals infected with HIV together with its complications In thi way the proposed technology meets NIHandapos s mission to treat immune mediated diseases to develop new and more effective treatments for HIV infected people and to find a cure Specifically the technology supports NIAID HIV Cure Research to determine the effects of latency reversal immune modulation cell therapies and other interventions on the HIV reservoir study HIV latency in the context of clinically infected whole blood cultures and investigate strategies that lead to purging of the latent reservoir and a cure for HIV AIDS or control of viral rebound and sustained remission