A scalable workflow to test “shock and kill” therapeutic approaches against the HIV-1 latent reservoir in blood cells ex vivo

Abstract

Summary: Integrated HIV-1 DNA persists in cells of people living with HIV during antiretroviral treatment, but its quantification is hindered by its rarity. Here, we present an optimized protocol to evaluate “shock and kill” therapeutic strategies, including both the latency reactivation (“shock”) and elimination of infected cells (“kill”) stages. We describe steps for the sequential use of nested PCR-based assays and viability sorting to allow for scalable and rapid screening of candidate therapeutics in patient-derived blood cells.For complete details on the use and execution of this protocol, please refer to Shytaj et al..1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Keywords

• Cell Biology
• Cell Isolation
• Cell-based Assays
• Cell Separation/Fractionation
• Immunology
• Microbiology