Integrated and Total HIV-1 DNA Predict Ex Vivo Viral Outgrowth

Kiselinova, Maja; Spiegelaere, Ward De; Buzón, María J.; Malatinková, Eva; Lichterfeld, Mathias; Vandekerckhove, Linos

doi:10.1371/journal.ppat.1005472

Cited by 81 publications

(98 citation statements)

References 45 publications

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“…When compared with alternative measurements of reservoir size, the HIV RNA/Gag assay most closely mirrors findings using the VOA, where the minimum size of the replication-competent reservoir is estimated to be 1 per million resting CD4 (1.4 in our cohort), 100–1000 fold less than that measured by integrated DNA. In addition, both the reservoir size as measured by the HIV RNA/Gag assay here and the VOA, shown previously (Kiselinova et al, 2016), correlate with the reservoir measured by integrated HIV DNA. The advantage of the HIV RNA/Gag assay is the direct detection of HIV-infected cells, allowing phenotyping, which is not possible with the VOA.…”

Section: Discussionsupporting

confidence: 77%

Single-Cell Characterization of Viral Translation-Competent Reservoirs in HIV-Infected Individuals

Baxter

Nießl

Fromentin

et al. 2016

Cell Host & Microbe

176

268

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SUMMARY HIV cure efforts are hampered by limited characterization of the cells supporting HIV replication in vivo and inadequate methods for quantifying the latent viral reservoir in patients receiving antiretroviral therapy. We combine fluorescent in situ RNA hybridization with detection of HIV protein and flow cytometry, enabling detection of 0.5–1 gag-pol mRNA+/Gag protein+ infected cells per million. In the peripheral blood of untreated persons, active HIV replication correlated with viremia, and occurred in CD4 T cells expressing T follicular helper cell markers and inhibitory co-receptors. In virally-suppressed subjects, the approach identified latently infected cells capable of producing HIV mRNA and protein after stimulation with PMA/ionomycin and latency-reversing agents (LRAs). While ingenol-induced reactivation mirrored the effector and central/transitional memory CD4 T cell contribution to the pool of integrated HIV DNA, bryostatin-induced reactivation occurred predominantly in cells expressing effector memory markers. This indicates that CD4 T cell differentiation status differentially affects LRA effectiveness.

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Section: Discussionsupporting

confidence: 77%

Single-Cell Characterization of Viral Translation-Competent Reservoirs in HIV-Infected Individuals

Baxter

Nießl

Fromentin

et al. 2016

Cell Host & Microbe

176

268

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“…Similar results were recently reported by Kiselinova et al (11). More recently, Noel et al also reported that the low level of total HIV DNA in cells from long-term nonprogressors (LTNPs) correlated with the low efficiency of virus production after activation ex vivo (58).…”

Section: Dna Load In Resting Cd4supporting

confidence: 86%

“…Two comprehensive studies have compared a panel of HIV reservoir biomarkers (9,11), but such studies are limited by the fact that large amounts of blood are needed to test all markers in a given patient. The different approaches may be used differently according to the issue in question, such as the overall level of HIV infection in the body, viral persistence, reservoir activity, or the role of the HIV reservoir in maintaining immune activation.…”

Section: Place Of Total Hiv Dna Among Markers Used To Evaluate Hiv Rementioning

confidence: 99%

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Total HIV-1 DNA, a Marker of Viral Reservoir Dynamics with Clinical Implications

et al. 2016

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SUMMARYHIV-1 DNA persists in infected cells despite combined antiretroviral therapy (cART), forming viral reservoirs. Recent trials of strategies targeting latent HIV reservoirs have rekindled hopes of curing HIV infection, and reliable markers are thus needed to evaluate viral reservoirs. Total HIV DNA quantification is simple, standardized, sensitive, and reproducible. Total HIV DNA load influences the course of the infection and is therefore clinically relevant. In particular, it is predictive of progression to AIDS and death, independently of HIV RNA load and the CD4 cell count. Baseline total HIV DNA load is predictive of the response to cART. It declines during cART but remains quantifiable, at a level that reflects both the history of infection (HIV RNA zenith, CD4 cell count nadir) and treatment efficacy (residual viremia, cumulative viremia, immune restoration, immune cell activation). Total HIV DNA load in blood is also predictive of the presence and severity of some HIV-1-associated end-organ disorders. It can be useful to guide individual treatment, notably, therapeutic de-escalation. Although it does not distinguish between replication-competent and -defective latent viruses, the total HIV DNA load in blood, tissues, and cells provides insights into HIV pathogenesis, probably because all viral forms participate in host cell activation and HIV pathogenesis. Total HIV DNA is thus a biomarker of HIV reservoirs, which can be defined as all infected cells and tissues containing all forms of HIV persistence that participate in pathogenesis. This participation may occur through the production of new virions, creating new cycles of infection and disseminating infected cells; maintenance or amplification of reservoirs by homeostatic cell proliferation; and viral transcription and synthesis of viral proteins without new virion production. These proteins can induce immune activation, thus participating in the vicious circle of HIV pathogenesis.

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“…Although measurement of total HIV-1 DNA levels may take into account many copies of replication-deficient viruses, it is likely that under conditions of very low levels of viral replication, as in HICs, differences in the levels of HIV-1 DNA probably maintain proportionality between replication-competent and noncompe-tent proviruses, as recently observed in treated patients (54). This assumption is supported, despite limitations in the total numbers of CD4 ϩ T cells tested, by the direct correlation between the levels of cell-associated HIV-1 DNA and HIV-1 reactivation, measured by determination of the levels of HIV-1 RNA or p24 production in culture supernatants, that was found when CD4 ϩ T cells were split into parallel cultures and were subjected to different repeated stimuli (with polyclonal or allogeneic stimuli or the provision of latency-reversing agents).…”

Section: Discussionmentioning

confidence: 99%

Long-Term Spontaneous Control of HIV-1 Is Related to Low Frequency of Infected Cells and Inefficient Viral Reactivation

Noël

Peña

David

et al. 2016

J Virol

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HIV establishes reservoirs of infected cells that persist despite effective antiretroviral therapy (ART). In most patients, the virus begins to replicate soon after treatment interruption. However, a low frequency of infected cells at the time of treatment interruption has been associated with delayed viral rebound. Likewise, individuals who control the infection spontaneously, so-called HIV-1 controllers (HICs), carry particularly low levels of infected cells. It is unclear, however, whether and how this small number of infected cells contributes to durable viral control. Here we compared 38 HICs with 12 patients on effective combined antiretroviral therapy (cART) and found that the low frequency of infected cells in the former subjects was associated both with less efficient viral reactivation in resting CD4؉ T cells and with less efficient virion production ex vivo. We also found that a potent HIV-specific CD8 ؉ T cell response was present only in those HICs whose CD4 ؉ T cells produced virus ex vivo. Long-term spontaneous control of HIV infection in HICs thus appears to be sustained on the basis of the inefficient reactivation of viruses from a limited number of infected cells and the capacity of HICs to activate a potent HIV-specific CD8؉ T cell response to counteract efficient viral reactivation events. IMPORTANCEThere is a strong scientific interest in developing strategies to eradicate the HIV-1 reservoir. Very rare HIV-1-infected patients are able to spontaneously control viremia for long periods of time (HIV-1 controllers [HICs]) and are put forward as a model of HIV-1 remission. Here, we show that the low viral reservoirs found in HICs are a critical part of the mechanisms underlying viral control and result in a lower probability of HIV-1 reactivation events, resulting in limited HIV-1 release and spread. We found that those HICs in whom viral reactivation and spread from CD4 ؉ T cells in vitro were the most difficult were those with diminished CD8 ؉ T cell responses. These results suggest that, in some settings, low HIV-1 reservoirs decisively contribute to at least the temporary control of infection without antiretroviral therapy. We believe that this work provides information of relevance in the context of the search for HIV-1 remission.

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Integrated and Total HIV-1 DNA Predict Ex Vivo Viral Outgrowth

Cited by 81 publications

References 45 publications

Single-Cell Characterization of Viral Translation-Competent Reservoirs in HIV-Infected Individuals

Single-Cell Characterization of Viral Translation-Competent Reservoirs in HIV-Infected Individuals

Total HIV-1 DNA, a Marker of Viral Reservoir Dynamics with Clinical Implications

Long-Term Spontaneous Control of HIV-1 Is Related to Low Frequency of Infected Cells and Inefficient Viral Reactivation

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