An In-Depth Comparison of Latency-Reversing Agent Combinations in Various In Vitro and Ex Vivo HIV-1 Latency Models Identified Bryostatin-1+JQ1 and Ingenol-B+JQ1 to Potently Reactivate Viral Gene Expression

• DOI: 10.1371/journal.ppat.1005063
• Publication Date: 2015-07-30T18:14:30Z
• Authors: Gilles Darcis; Anna Kula; Sophie Bouchat; Koh Fujinaga; Francis Corazza; Amina Ait-Ammar; Nadège Delacourt; Adeline Melard; Kabamba Kabeya; Caroline Vanhulle; Benoit Van Driessche; Jean-Stéphane Gatot; Thomas Cherrier; Luiz F. Pianowski; Lucio Gama; Christian Schwartz; Jorge Vila; Arsène Burny; Nathan Clumeck; Michel Moutschen; Stéphane De Wit; B. Matija Peterlin; Christine Rouzioux; Olivier Rohr; Carine Van Lint
• Keywords: Ex vivo; P-TEFb; Bryostatin 1; Virus latency; Priming (agriculture)

The persistence of latently infected cells in patients under combinatory antiretroviral therapy (cART) is a major hurdle to HIV-1 eradication. Strategies purge these reservoirs are needed and activation viral gene expression one promising strategy. Bromodomain Extraterminal (BET) bromodomain inhibitors (BETi) compounds able reactivate latent proviruses positive transcription elongation factor b (P-TEFb)-dependent manner. In this study, we tested the reactivation potential protein kinase C (PKC) agonists (prostratin, bryostatin-1 ingenol-B), which known activate NF-κB signaling pathway as well P-TEFb, used alone or combination with P-TEFb-releasing agents (HMBA BETi (JQ1, I-BET, I-BET151)). Using vitro post-integration latency model cell lines T-lymphoid myeloid lineages, demonstrated that PKC acted potent latency-reversing (LRAs) their combinations led synergistic at mRNA levels. Mechanistically, combined treatments higher activations P-TEFb than corresponding individual drug treatments. Importantly, observed ex vivo cultures CD8+-depleted PBMCs from 35 cART-treated HIV-1+ aviremic percentage reactivated following bryostatin-1+JQ1 treatment was identical anti-CD3+anti-CD28 antibodies control stimulation. Remarkably, resting CD4+ T isolated 15 patients, ingenol-B+JQ1 released infectious viruses levels similar obtained effects two were already detected 24 hours post-stimulation. These results constitute first demonstration LRA exhibiting such effect represent proof-of-concept for co-administration different types LRAs strategy reduce size reservoirs.