Latency reversal agents affect differently the latent reservoir present in distinct CD4+ T subpopulations

• DOI: 10.1371/journal.ppat.1007991
• Publication Date: 2019-08-19T13:32:24Z
• Authors: Judith Grau-Expósito; Laura Luque-Balle...; Jordi Navarro; Adrian Curran; Joaquin Burgos; Esteban Ribera; Ariadna Torrella; Bibiana Planas; Rosa Badía; Mario Martin-Cast...; Jesús Fernández-Sojo; Meritxell Genescà; Vicenç Falcó; Maria J. Buzon
• Keywords: CD4-Positive T-Lymphocytes; QH301-705.5; Anti-HIV Agents; :Hemic and Immune Systems::Blood::Blood Cells::Leukocytes::Leukocytes, Mononuclear::Lymphocytes::T-Lymphocytes::Hemic and Immune Systems::CD4-Positive T-Lymphocytes [ANATOMY]; HIV Infections; :Microbiological Phenomena::Virus Physiological Phenomena::Virus Latency [PHENOMENA AND PROCESSES]; 03 medical and health sciences; Depsipeptides; Humans; Biology (General); :acciones y usos químicos::acciones farmacológicas::usos terapéuticos::antiinfecciosos::antivíricos::antirretrovirales::fármacos anti-VIH [COMPUESTOS QUÍMICOS Y DROGAS]; 0303 health sciences; Virus - Reproducció; Antiretrovirals; :Chemical Actions and Uses::Pharmacologic Actions::Therapeutic Uses::Anti-Infective Agents::Antiviral Agents::Anti-Retroviral Agents::Anti-HIV Agents [CHEMICALS AND DRUGS]; RC581-607; Viral Load; Virus Latency; 3. Good health; :sistemas sanguíneo e inmunológico::sistemas sanguíneo e inmunológico::sistema inmunológico::leucocitos::leucocitos mononucleares::linfocitos::linfocitos T::linfocitos T CD4-positivos [ANATOMÍA]; Cèl·lules T; HIV-1; Virus Activation; Immunologic diseases. Allergy; Diterpenes; :fenómenos microbiológicos::fenómenos fisiológicos de los virus::latencia viral [FENÓMENOS Y PROCESOS]; Research Article

Latency reversal agents (LRAs) have proven to induce HIV-1 transcription in vivo but are ineffective at decreasing the size of the latent reservoir in antiretroviral treated patients. The capacity of the LRAs to perturb the viral reservoir present in distinct subpopulations of cells is currently unknown. Here, using a new RNA FISH/flow ex vivo viral reactivation assay, we performed a comprehensive assessment of the viral reactivation capacity of different families of LRAs, and their combinations, in different CD4+ T cell subsets. We observed that a median of 16.28% of the whole HIV-reservoir induced HIV-1 transcripts after viral reactivation, but only 10.10% of these HIV-1 RNA+ cells produced the viral protein p24. Moreover, none of the LRAs were powerful enough to reactivate HIV-1 transcription in all CD4+ T cell subpopulations. For instance, the combination of Romidepsin and Ingenol was identified as the best combination of drugs at increasing the proportion of HIV-1 RNA+ cells, in most, but not all, CD4+ T cell subsets. Importantly, memory stem cells were identified as highly resistant to HIV-1 reactivation, and only the combination of Panobinostat and Bryostatin-1 significantly increased the number of cells transcribing HIV within this subset. Overall, our results validate the use of the RNA FISH/flow technique to assess the potency of LRAs among different CD4+ T cell subsets, manifest the intrinsic differences between cells that encompass the latent HIV reservoir, and highlight the difficulty to significantly impact the latent infection with the currently available drugs. Thus, our results have important implications for the rational design of therapies aimed at reversing HIV latency from diverse cellular reservoirs.