Quantifying integrated SIV-DNA by repetitive-sampling Alu-gag PCR.

• Type of Material: Article
• Authors: Maud Mavigner, Emory University; S. Thera Lee, Emory University; Jakob Habib, Emory University; Cameron Robinson, Emory University; Guido Silvestri, Emory University; Una O'Doherty, University of Pennsylvania; Ann Chahroudi, Emory University
• Language: English
• Date: 2016-10-05
• Publisher: Mediscript Ltd
• Publication Version: Final Published Version
• Copyright Statement: © 2016 The Authors.
• License: Creative Commons Attribution-NonCommercial 4.0 International
• Title of Journal or Parent Work: Journal of Virus Eradication
• Volume: 2
• Issue: 4
• Start Page: 219
• End Page: 226
• Grant/Funding Information: This work was partially supported by the amfAR Innovation Grant Program 109353-59-RGRL to MM. AC acknowledges funding from the National Center for Advancing Translational Sciences of the National Institutes of Health under Award UL1TR000454 and Support of the Yerkes National Primate Research Center (P51 OD011132).
• Keywords: reservoir; rhesus macaques; integration; latency; SIV
• Research Categories: Biology, Virology; Health Sciences, Pathology

Abstract

OBJECTIVES: Although antiretroviral therapy (ART) effectively suppresses HIV-1 replication, it does not eradicate the virus and ART interruption consistently results in rebound of viraemia, demonstrating the persistence of a long-lived viral reservoir. Several approaches aimed at reducing virus persistence are being developed, and accurate measurements of the latent reservoir (LR) are necessary to assess the effectiveness of anti-latency interventions. We sought to measure the LR in SIV/SHIV-infected rhesus macaques (RMs) by quantifying integrated SIV-DNA. METHODS: We optimised a repetitive sampling Alu-gag PCR to quantify integrated SIV-DNA ex vivo in ART-naïve and ART-experienced SIV/SHIV-infected RMs. RESULTS: In ART-naïve RMs, we found the median level of integrated SIV-DNA to be 1660 copies and 866 copies per million PBMC during untreated acute and chronic SHIV infection, respectively. Integrated and total SIV-DNA levels were positively correlated with one another. In ART-treated RMs, integrated SIV-DNA was readily detected in lymph nodes and spleen and levels of total (3319 copies/million cells) and integrated (3160 copies/million cells) SIV-DNA were similar after a median of 404 days of ART. In peripheral blood CD4+ T cells from ART-treated RMs, levels of total (3319 copies/million cells) and integrated (2742 copies/million cells) SIV-DNA were not significantly different and were positively correlated. CONCLUSIONS: The assay described here is validated and can be used in interventional studies testing HIV/SIV cure strategies in RMs. Measurement of integrated SIV-DNA in ART-treated RMs, along with other reservoir analyses, gives an estimate of the size of the LR.