A Tat/Rev Induced Limiting Dilution Assay to Measure Viral Reservoirs in Non-Human Primate Models of HIV Infection

• Type of Material: Article
• Authors: Ines Frank, Population Council; Arpan Acharya, University of Nebraska; Nanda K. Routhu, Emory University; Meropi Aravantinou, Population Council; Justin L. Harper, Emory University; Stephanie Maldonado, Population Council; Maria Sole Cigoli, Population Council; Stanka Semova, Rockefeller University; Svetlana Mazel, Rockefeller University; Mirko Paiardini, Emory University; Nina Derby, Population Council; Siddappa Byrareddy, Emory University; Elena Martinelli, Population Council
• Language: English
• Date: 2019-08-19
• Publisher: Nature Research (part of Springer Nature): Fully open access journals
• Publication Version: Final Published Version
• Copyright Statement: © 2019, The Author(s).
• Final Published Version (URL): http://dx.doi.org/10.1038/s41598-019-48354-3
• Title of Journal or Parent Work: Scientific Reports
• ISSN: 2045-2322
• Volume: 9
• Issue: 1
• Start Page: 12078
• End Page: 12078
• Grant/Funding Information: This work was supported in part by NIH grants R56AI098546-06, R01AI040877-20, R01AI129745; R21MH113455; P30MH062261, ORIP/OD P51OD011132.
• Keywords: TRANSCRIPTION; Science & Technology; LYMPHOCYTES; SIZE; Science & Technology - Other Topics; Multidisciplinary Sciences; ANTIRETROVIRAL THERAPY; LATENT RESERVOIR; MACAQUES; CELL; GENE-EXPRESSION; PERSISTENCE; SIMIAN IMMUNODEFICIENCY VIRUS
• Research Categories: Health Sciences, Immunology

Abstract

The establishment of latent infection and poorly characterized viral reservoirs in tissues represent major obstacles to a definitive cure for HIV. Non-human primate (NHP) models of HIV infection are critical to elucidate pathogenic processes and an essential tool to test novel therapeutic strategies. Thus, the availability of novel assays to measure residual viral replication and reservoirs in NHP models may increase their utility in the search for an HIV cure. We developed a tat/rev induced limiting dilution assay to measure the frequency of CD4+ T cells that express multiply-spliced(ms)_SIV RNA in presence and absence of stimulation. We validated the assay using cell lines and cells from blood and lymph nodes of SIV infected macaques. In vitro, SIV/SHIV TILDA detects only cells expressing viral proteins. In SIV/SHIV-infected macaques, CD4+ T cells that express msSIV/SHIV RNA (TILDA data) were detected also in the setting of very low/undetectable viremia. TILDA data were significantly higher after stimulation and correlated with plasma viral load (pVL). Interestingly, TILDA data from early cART initiation correlated with peak and AUC pVL post-cART interruption. In summary, we developed an assay that may be useful in characterizing viral reservoirs and determining the effect of HIV interventions in NHP models.