A Murine Viral Outgrowth Assay to Detect Residual HIV Type 1 in Patients With Undetectable Viral Loads

Pate, Kelly A. Metcalf; Pohlmeyer, Christopher W.; Walker-Sperling, Victoria E.; Foote, Jeremy B.; Najarro, Kevin M.; Cryer, Catherine G.; Salgado, María; Gama, Lúcio; Engle, Elizabeth L.; Shirk, Erin N.; Queen, Suzanne E.; Chioma, Stanley; Vermillion, Meghan S.; Bullock, Brandon; Li, Ming; Lyons, Claire E.; Adams, Robert J.; Zink, M. Christine; Clements, Janice E.; Mankowski, Joseph L.; Blankson, Joel N.

doi:10.1093/infdis/jiv230

Cited by 57 publications

(64 citation statements)

References 39 publications

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“…However, a major hurdle impeding more widespread use of humanized mice is that each experiment requires the surgical engraftment of human tissue, since this aspect cannot be bred [124]. A promising and creative use of humanized mice is in the development of a murine viral outgrowth assay where HIV-1 latency is estimated by adoptive transfer of human cells into humanized mice [131]. …”

Section: Biologic Solutionsmentioning

confidence: 99%

Are T cells the only HIV-1 reservoir?

2016

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Current antiretroviral therapies have improved the duration and quality of life of people living with HIV-1. However, viral reservoirs impede complete eradication of the virus. Although there are many strategies to eliminate infectious virus, the most actively pursued are latency reversing agents in conjunction with immune modulation. This strategy, known as “shock and kill”, has been tested primarily against the most widely recognized HIV-1 latent reservoir found in resting memory CD4+ T cells. This is in part because of the dearth of conclusive evidence about the existence of non-T cell reservoirs. Studies of non-T cell reservoirs have been difficult to interpret because of technical and biological issues that have hampered a better understanding. This review considers the current knowledge of non-T cell reservoirs, the challenges encountered in a better understanding of these populations, and their implications for HIV-1 cure research.

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Section: Biologic Solutionsmentioning

confidence: 99%

Are T cells the only HIV-1 reservoir?

2016

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“…The assay is labour intensive, expensive and requires large numbers of cells. Recent advances in measuring inducible virus have included a similar limiting dilution format but measuring production of cell associated RNA or release of viral RNA in supernatant 87,88 and amplification of latent infectious virus using a humanized mouse model – the murine viral outgrowth assay (MVOA) 89 .…”

Section: Novel Biomarkers To Analyse/quantify Hiv Reservoirsmentioning

confidence: 99%

“…Early initiation of ART limits the establishment of the reservoir and prevents the generation of immune escape in latently infected cells 63,117,118 . New tools that can quantify the frequency of a cell that carries replication competent virus have been developed 88,119 , and some biomarkers have been shown to predict the time to viral rebound following a treatment interruption 92,93,120,121 . The central role of the T follicular helper cells and the B cell follicle in supporting SIV/HIV replication was established 35 .…”

Section: Figurementioning

confidence: 99%

International AIDS Society global scientific strategy: towards an HIV cure 2016

Deeks¹,

Lewin²,

Ross³

et al. 2016

Nat Med

388

348

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Antiretroviral therapy is not curative. Given the challenges in providing life-long therapy to a global population of over 35 million people living with HIV, there is intense interest in developing a cure for HIV infection. The International AIDS Society convened a group of international experts to develop a scientific strategy for research towards an HIV cure. This Perspective summarizes the group's strategy.

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“…The murine viral outgrowth assay (MVOA) is a binary end-point assay that uses a mouse model to determine whether patient-derived cells harbor infectious virus [78]. In this assay, either whole PBMCs or sorted CD4 T cells are injected into NOD/Prkdc scid /gamma-chain knockout (NSG) mice.…”

Section: Introductionmentioning

confidence: 99%

“…Over time, HIV RNA from the plasma of xenografted mice is isolated and quantified by RT-qPCR. The assay was successful in recovering virus from patient cells, including an elite controller, who had negative QVOA results [78]. It can be used to survey a large number of patient cells, requiring 1 mouse per 10-50 million CD4 T cells.…”

Section: Introductionmentioning

confidence: 99%

The Alphabet Soup of HIV Reservoir Markers

Sharaf

2017

Curr HIV/AIDS Rep

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Purpose of review Despite the success of antiretroviral therapy in suppressing HIV, life-long therapy is required to avoid HIV reactivation from long-lived viral reservoirs. Currently, there is intense interest in searching for therapeutic interventions that can purge the viral reservoir to achieve complete remission in HIV patients off antiretroviral therapy. The evaluation of such interventions relies on our ability to accurately and precisely measure the true size of the viral reservoir. In this review, we assess the most commonly used HIV reservoir assays, as a clear understanding of the strengths and weaknesses of each is vital for the accurate interpretation of results and for the development of improved assays. Recent findings The quantification of intracellular or plasma HIV RNA or DNA levels remains the most commonly used tests for the characterization of the viral reservoir. While cost-effective and high-throughput, these assays are not able to differentiate between replication-competent or defective fractions, or quantify the number of infected cells. Viral outgrowth assays provide a lower bound for the fraction of cells that can produce infectious virus, but these assays are laborious, expensive and substantially underestimate the potential reservoir of replication-competent provirus. Newer assays are now available that seek to overcome some of these problems, including full-length proviral sequencing, inducible HIV RNA assays, ultrasensitive p24 assays and murine adoptive transfer techniques. Summary The development and evaluation of strategies for HIV remission relies upon our ability to accurately and precisely quantify the size of the remaining viral reservoir. At this time, all current HIV reservoir assays have drawbacks such that combinations of assays are generally needed to gain a more comprehensive view of the viral reservoir. The development of novel, rapid, high-throughput assays that can sensitively quantify the levels of the replication-competent HIV reservoir is still needed.

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A Murine Viral Outgrowth Assay to Detect Residual HIV Type 1 in Patients With Undetectable Viral Loads

Abstract: Our results suggest that the MVOA has the potential to serve as a powerful tool to identify residual HIV in patients with undetectable viral loads.

Cited by 57 publications

References 39 publications

Are T cells the only HIV-1 reservoir?

Are T cells the only HIV-1 reservoir?

International AIDS Society global scientific strategy: towards an HIV cure 2016

The Alphabet Soup of HIV Reservoir Markers

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