Quantifying integrated SIV-DNA by repetitive-sampling Alu-gag PCR
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 pers...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2016-10-01
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| Series: | Journal of Virus Eradication |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2055664020308700 |
| _version_ | 1819049410913370112 |
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| author | Maud Mavigner S. Thera Lee Jakob Habib Cameron Robinson Guido Silvestri Una O’Doherty Ann Chahroudi |
| author_facet | Maud Mavigner S. Thera Lee Jakob Habib Cameron Robinson Guido Silvestri Una O’Doherty Ann Chahroudi |
| author_sort | Maud Mavigner |
| collection | DOAJ |
| description | 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-gagPCR 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. |
| first_indexed | 2024-12-21T11:31:43Z |
| format | Article |
| id | doaj.art-51612f8cd7e04f7fa4f1b52e329bb19f |
| institution | Directory Open Access Journal |
| issn | 2055-6640 |
| language | English |
| last_indexed | 2024-12-21T11:31:43Z |
| publishDate | 2016-10-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of Virus Eradication |
| spelling | doaj.art-51612f8cd7e04f7fa4f1b52e329bb19f2022-12-21T19:05:32ZengElsevierJournal of Virus Eradication2055-66402016-10-0124219226Quantifying integrated SIV-DNA by repetitive-sampling Alu-gag PCRMaud Mavigner0S. Thera Lee1Jakob Habib2Cameron Robinson3Guido Silvestri4Una O’Doherty5Ann Chahroudi6Yerkes National Primate Research Center, Emory University, Atlanta, GA, USA; Department of Pediatrics, Emory University School of Medicine, Atlanta, GA, USAYerkes National Primate Research Center, Emory University, Atlanta, GA, USA; Department of Pediatrics, Emory University School of Medicine, Atlanta, GA, USADepartment of Pediatrics, Emory University School of Medicine, Atlanta, GA, USAYerkes National Primate Research Center, Emory University, Atlanta, GA, USAYerkes National Primate Research Center, Emory University, Atlanta, GA, USADepartment of Pathology and Laboratory Medicine, University of Pennsylvania, Philadelphia, PA, USAYerkes National Primate Research Center, Emory University, Atlanta, GA, USA; Department of Pediatrics, Emory University School of Medicine, Atlanta, GA, USA; Corresponding author: Ann Chahroudi, E472, HSRB, 1760 Haygood Drive, Atlanta, GA, 30322, USAObjectives: 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-gagPCR 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.http://www.sciencedirect.com/science/article/pii/S2055664020308700SIVintegrationreservoirlatencyrhesus macaques |
| spellingShingle | Maud Mavigner S. Thera Lee Jakob Habib Cameron Robinson Guido Silvestri Una O’Doherty Ann Chahroudi Quantifying integrated SIV-DNA by repetitive-sampling Alu-gag PCR Journal of Virus Eradication SIV integration reservoir latency rhesus macaques |
| title | Quantifying integrated SIV-DNA by repetitive-sampling Alu-gag PCR |
| title_full | Quantifying integrated SIV-DNA by repetitive-sampling Alu-gag PCR |
| title_fullStr | Quantifying integrated SIV-DNA by repetitive-sampling Alu-gag PCR |
| title_full_unstemmed | Quantifying integrated SIV-DNA by repetitive-sampling Alu-gag PCR |
| title_short | Quantifying integrated SIV-DNA by repetitive-sampling Alu-gag PCR |
| title_sort | quantifying integrated siv dna by repetitive sampling alu gag pcr |
| topic | SIV integration reservoir latency rhesus macaques |
| url | http://www.sciencedirect.com/science/article/pii/S2055664020308700 |
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