Automated Multireplicate Quantification of Persistent HIV-1 Viremia in Individuals on Antiretroviral Therapy

Jacobs, Jana L.; Tosiano, Melissa A.; Koontz, Dianna; Staines, Brittany; Worlock, Andrew; Harrington, Karen; Bakkour, Sonia; Stone, Mars; Shutt, Kathleen A.; Busch, Michael P.; Mellors, John W.

doi:10.1128/jcm.01442-20

Cited by 13 publications

(10 citation statements)

References 23 publications

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“…The single copy RNA assay has shown that most clinically suppressed patients have persistent low-level viremia but it has only been used in research and clinical trial contexts [18,21,[23][24][25][26] and has the disadvantage of a limited range from 0 to 20 copies. The Double-R assay has an improved dynamic range of at least 3 log 10 copies making it more suitable for routine clinical monitoring.…”

Section: Discussionmentioning

confidence: 99%

HIV-1 viral blips are associated with repeated and increasingly high levels of cell-associated HIV-1 RNA transcriptional activity

Suzuki¹,

Levert-Mignon²,

Yeung³

et al. 2021

Aids

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Objective: Some HIV+ patients, virally suppressed on ART, show occasional ‘blips’ of detectable HIV-1 plasma RNA. We used a new highly sensitive assay of cell-associated HIV-1 RNA to measure transcriptional activity in PBMCs and production of infectious virus from the viral reservoir, in patients with and without ‘blips’. Design/methods: RNA and DNA extracted from cells in 6 ml of peripheral blood, from suppressed patients with one to two ‘blip’ episodes over the past 2 years of ART ( n = 55), or no ‘blips’ ( n = 52), were assayed for HIV-1 RNA transcripts and proviral DNA targeting the highly conserved ‘R’ region of the LTR. Follow-up samples were also collected. Purified CD4 + T cells were cultured with anti-CD3/CD28/CD2 T-cell activator to amplify transcription and measure replication competent virus. Results: HIV-1 RNA transcripts ranged from 1.3 to 5415 copies/10 6 white blood cells. ‘Blip’ patients had significantly higher levels vs. without blips (median 192 vs. 49; P = 0.0007), which correlated with: higher levels of inducible transcripts after activation in vitro , sustained higher HIV-1 transcription levels in follow-up samples along with increasing HIV-1 DNA in some, and production of replication-competent HIV-1. Conclusion: Viral ‘blips’ are significant reflecting higher transcriptional activity from the reservoir and contribute to the reservoir over time. This sensitive assay can be used in monitoring the size and activity of the HIV-1 reservoir and will be useful in HIV-1 cure strategies.

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Section: Discussionmentioning

confidence: 99%

HIV-1 viral blips are associated with repeated and increasingly high levels of cell-associated HIV-1 RNA transcriptional activity

Suzuki¹,

Levert-Mignon²,

Yeung³

et al. 2021

Aids

View full text Add to dashboard Cite

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“…This could explain why viral reservoir size and the emergence of pLLV were not correlated by Widera et al (2017) [ 132 ]. Moreover, from the studies reviewed, Jacobs et al were unable to identify a cellular or titular reservoir contributing to persistent viremia and thus proposed proviral replication as the main cause of the continuous presence of the virus [ 133 ].…”

Section: Non-aids Defining Events (Nades) and Pllvmentioning

confidence: 99%

Persistent low-Level viremia in persons living with HIV undertreatment: An unresolved status

et al. 2021

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Antiretroviral therapy (ART) allows suppressed viremia to reach less than 50 copies/mL in most treated persons living with HIV (PLWH). However, the existence of PLWH that show events of persistent low-level viremia (pLLV) between 50 and 1000 copies/mL and with different virological consequences have been observed. PLLV has been associated with higher virological failure (VF), viral genotype resistance, adherence difficulties and AIDS events. Moreover, some reports show that pLLV status can lead to residual immune activation and inflammation, with an increased risk of immunovirological failure and a pro-inflammatory cytokine level which can lead to a higher occurrence of non-AIDS defining events (NADEs) and other adverse clinical outcomes. Until now, however, published data have shown controversial results that hinder understanding of the true cause(s) and origin(s) of this phenomenon. Molecular mechanisms related to viral reservoir size and clonal expansion have been suggested as the possible origin of pLLV. This review aims to assess recent findings to provide a global view of the role of pLLV in PLWH and the impact this status may cause on the clinical progression of these patients.

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“…These studies used relatively fewer reps (9 to 18 reps) than reported in the current study, but this difference had only a small impact on the rates of detection and estimated viral loads linked to interventions. Furthermore, Jacobs et al evaluated the replicate Aptima assay using 9 reps in comparison to the manual single-copy assay targeting integrase (iSCA v2), showing comparable sensitivity ( 30 , 31 ).…”

Section: Discussionmentioning

confidence: 99%

Replicate Aptima Assay for Quantifying Residual Plasma Viremia in Individuals on Antiretroviral Therapy

et al. 2020

Self Cite

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Detection of residual plasma viremia in ART-suppressed HIV-infected individuals is critical for characterizing the latent reservoir and evaluating the impact of cure interventions. Ultracentrifugation-based single-copy assays are sensitive but labor-intensive. Fully automated replicate testing using a standard clinical viral load assay was evaluated as a high-throughput alternative for quantification of low-level viremia. Four plasma samples from blood donors with acute HIV-1 infection and one viral culture supernatant were serially diluted into 25 mL samples to nominal viral loads ranging from 39 to <0.5 copies (cp)/mL. Each dilution was tested with 45 replicates (reps) using 0.5 mL/rep with the Aptima HIV-1 Quant assay. The nominal and estimated viral loads based on the single-hit Poisson model were compared, and a hybrid Poisson digital model for calibrated viral load estimation was derived. Testing performed using 45 reps on longitudinal plasma samples from 50 ART-suppressed individuals in the Reservoir Assay Validation and Evaluation Network (RAVEN) study cohort (range 1-19 years of continuous ART suppression) showed a median viral load of 0.54 cp/mL (IQR 0.22-1.46 cp/mL) and a 14% (95% CI 9-19%) decline in viral load for each additional year in duration suppressed. Within the RAVEN cohort, the expected false negative rate for detection at lower rep numbers using 9 and 18 reps was 26% and 14%, respectively. Residual plasma viremia levels positively correlated with cell-associated HIV RNA and DNA. The performance characteristics of the replicate Aptima assay support its use for quantifying residual plasma viremia to study the latent HIV reservoir and cure interventions.

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Automated Multireplicate Quantification of Persistent HIV-1 Viremia in Individuals on Antiretroviral Therapy

Cited by 13 publications

References 23 publications

HIV-1 viral blips are associated with repeated and increasingly high levels of cell-associated HIV-1 RNA transcriptional activity

HIV-1 viral blips are associated with repeated and increasingly high levels of cell-associated HIV-1 RNA transcriptional activity

Persistent low-Level viremia in persons living with HIV undertreatment: An unresolved status

Replicate Aptima Assay for Quantifying Residual Plasma Viremia in Individuals on Antiretroviral Therapy

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