Intensification of ART with ABX464 decreases the total HIV reservoir and HIV transcription initiation in CD4 T cells from HIV-infected ART-suppressed individuals

Abstract Background Antiretroviral therapy (ART) intensification and disruption of latency have been suggested as strategies to eradicate HIV. ABX464 is a novel antiviral that inhibits HIV RNA biogenesis. We investigated the effect of ABX464 on HIV transcription and total and intact HIV DNA in CD4 + T cells from ART-suppressed participants enrolled in the ABIVAX-005 clinical trial (NCT02990325). Methods Peripheral CD4 + T cells were available for analysis from nine ART-suppressed participants who were treated daily with 150mg of ABX464 for 4 weeks. Total and intact HIV DNA, and initiated, 5’elongated, unspliced, polyadenylated and multiply-spliced HIV transcripts, were quantified at weeks 0, 4 and 8 using droplet digital PCR. Results We observed a significant decrease in total HIV DNA (p=0.008, median fold-change=0.8) and a lower median level of intact HIV DNA (p=n.s., median fold-change=0.8) after ABX464 treatment (wk 0 vs. 4). Moreover, we observed a decrease in initiated HIV RNA per million CD4 + T cells and per provirus (p=0.05, median fold-change=0.7; p=0.004, median fold-change=0.5, respectively), a trend towards a decrease in the 5’elongated HIV RNA per provirus (p=0.07, median fold-change=0.5), and a lower median level of unspliced HIV RNA (p=n.s., median fold-change=0.6), but no decrease in polyadenylated or multiply-spliced HIV RNA. Conclusion In this substudy, ABX464 had a dual effect of decreasing total HIV DNA (and possibly intact proviruses) and decreasing the amount of HIV transcription per provirus. To further characterize its specific mechanism of inhibiting HIV transcription, long-term administration of ABX464 should be studied in a larger cohort.

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Pharmacological Inhibition of PPARy Boosts HIV Reactivation and Th17 Effector Functions, while Preventing Progeny Virion Release and de novo Infection

Pathogens and Immunity ◽

10.20411/pai.v5i1.348 ◽

2020 ◽

Vol 5 (1) ◽

pp. 177

Author(s):

Delphine Planas ◽

Augustine Fert ◽

Yuwei Zhang ◽

Jean-Philippe Goulet ◽

Jonathan Richard ◽

...

Keyword(s):

T Cells ◽

Cd4 T Cells ◽

Th17 Cells ◽

People Living With Hiv ◽

Hiv Reservoir ◽

Effector Functions ◽

Pharmacological Inhibition ◽

Hiv Transcription ◽

Virion Release ◽

Progeny Virion

The frequency and functions of Th17-polarized CCR6+RORyt+CD4+ T cells are rapidly compromised upon HIV infection and are not restored with long-term viral suppressive antiretroviral therapy (ART). In line with this, Th17 cells represent selective HIV-1 infection targets mainly at mucosal sites, with long-lived Th17 subsets carrying replication-competent HIV-DNA during ART. Therefore, novel Th17-specific therapeutic interventions are needed as a supplement of ART to reach the goal of HIV remission/cure. Th17 cells express high levels of peroxisome proliferator-activated receptor gamma (PPARy), a transcriptional factor that represses the transcription of the HIV provirus and the rorc gene, which encodes for the Th17-specific master regulator RORyt/RORC2. Thus, we hypothesized that the pharmacological inhibition of PPARy will facilitate HIV reservoir reactivation while enhancing Th17 effector functions. Consistent with this prediction, the PPARy antagonist T0070907 significantly increased HIV transcription (cell-associated HIV-RNA) and RORyt-mediated Th17 effector functions (IL-17A). Unexpectedly, the PPARy antagonism limited HIV outgrowth from cells of ART-treated people living with HIV (PLWH), as well as HIV replication in vitro. Mechanistically, PPARy inhibition in CCR6+CD4+ T cells induced the upregulation of transcripts linked to Th17-polarisation (RORyt, STAT3, BCL6 IL-17A/F, IL-21) and HIV transcription (NCOA1-3, CDK9, HTATIP2). Interestingly, several transcripts involved in HIV-restriction were upregulated (Caveolin-1, TRIM22, TRIM5α, BST2, miR-29), whereas HIV permissiveness transcripts were downregulated (CCR5, furin), consistent with the decrease in HIV outgrowth/replication. Finally, PPARy inhibition increased intracellular HIV-p24 expression and prevented BST-2 downregulation on infected T cells, suggesting that progeny virion release is restricted by BST-2-dependent mechanisms. These results provide a strong rationale for considering PPARy antagonism as a novel strategy for HIV-reservoir purging and restoring Th17-mediated mucosal immunity in ART-treated PLWH.

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OA1-2 Enrichment of the HIV reservoir in CD32 + CD4 T cells occurs early and is closely associated with immune checkpoint receptor expression

Journal of Virus Eradication ◽

10.1016/s2055-6640(20)30834-7 ◽

2017 ◽

Vol 3 ◽

pp. 1-2

Author(s):

G.E. Martin ◽

M. Pace ◽

J.P. Thornhill ◽

C. Phetsouphanh ◽

E. Hopkins ◽

...

Keyword(s):

T Cells ◽

Immune Checkpoint ◽

Cd4 T Cells ◽

Receptor Expression ◽

Hiv Reservoir

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Flow Cytometry Sorting of Memory CCR6+CD4+ T-Cells for HIV Reservoir Quantification

Methods in Molecular Biology - HIV Reservoirs ◽

10.1007/978-1-0716-1871-4_7 ◽

2022 ◽

pp. 81-89

Author(s):

Amélie Cattin ◽

Augustine Fert ◽

Delphine Planas ◽

Petronela Ancuta

Keyword(s):

Flow Cytometry ◽

T Cells ◽

Cd4 T Cells ◽

Hiv Reservoir ◽

Flow Cytometry Sorting

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Impact of the level of pre-ART CD4+ T cells in blood on the rectal HIV reservoir in long-term treated men (VIRECT study)

BMC Infectious Diseases ◽

10.1186/1471-2334-14-s2-o17 ◽

2014 ◽

Vol 14 (S2) ◽

Author(s):

Delphine Vergnon-Miszczycha ◽

Alexandre Girard ◽

Anne Depincé ◽

Xavier Roblin ◽

Emilie Del Tedesco ◽

...

Keyword(s):

T Cells ◽

Cd4 T Cells ◽

Hiv Reservoir

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Infusion of CCR5 Gene-Edited T Cells Allows Immune Reconstitution, HIV Reservoir Decay, and Long-Term Virological Control

10.1101/2021.02.28.433290 ◽

2021 ◽

Author(s):

Joumana Zeidan ◽

Ashish A Sharma ◽

Gary Lee ◽

Angie Raad ◽

Remi Fromentin ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

Cd4 T Cells ◽

Ccr5 Gene ◽

Partial Control ◽

Hiv Reservoir ◽

Differentiated Cells ◽

Cell Counts ◽

Post Infusion

Antiretroviral therapy (ART) fails to fully restore immune function and is not curative. A single infusion of CCR5 gene-edited autologous CD4+ T cells (SB-728-T) led to sustained increases in CD4+ T cell counts, improved T cell homeostasis, and reduced the estimated size of the HIV reservoir. These outcomes were associated with the expansion and long-term persistence of a novel CCR5 gene-edited CD4+ T memory stem cell (CD45RAintROint TSCM) subset that can replenish the pool of more differentiated memory cells. We showed that novel CD45RAintROint TSCM cells are transcriptionally distinct from the previously described CD45RA+ TSCM and are minimally differentiated cells uncommitted to a specific Th-lineage. Subsequently, we showed in an independent trial that infusion of the SB-728-T cell product resulted in partial control of viral replication upon cessation of ART which was correlated with the frequencies of CCR5 gene-edited TSCM and their TEM progeny. Interestingly, one participant that remained off ART to this date demonstrated long-term maintenance of CCR5 gene-edited cells and increased frequency of polyfunctional HIV-specific CD4+ and CD8+ T cells, contributing to low levels of viral load 5 years post-infusion. Consequently, the generation of HIV protected memory CD4+ T cells by CCR5 disruption can contribute toward novel interventions aimed at achieving a sustained ART-free viral remission of HIV disease.

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Targeting HIV Reservoir in Infected CD4 T Cells by Dual-Affinity Re-targeting Molecules (DARTs) that Bind HIV Envelope and Recruit Cytotoxic T Cells

PLoS Pathogens ◽

10.1371/journal.ppat.1005233 ◽

2015 ◽

Vol 11 (11) ◽

pp. e1005233 ◽

Cited By ~ 61

Author(s):

Derek D. Sloan ◽

Chia-Ying Kao Lam ◽

Alivelu Irrinki ◽

Liqin Liu ◽

Angela Tsai ◽

...

Keyword(s):

T Cells ◽

Cd4 T Cells ◽

Cytotoxic T Cells ◽

Hiv Reservoir ◽

Hiv Envelope

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Impact of Biological Sex on Immune Activation and Frequency of the Latent HIV Reservoir During Suppressive Antiretroviral Therapy

The Journal of Infectious Diseases ◽

10.1093/infdis/jiaa298 ◽

2020 ◽

Vol 222 (11) ◽

pp. 1843-1852 ◽

Cited By ~ 3

Author(s):

Shane D Falcinelli ◽

Bonnie E Shook-Sa ◽

Morgan G Dewey ◽

Sumati Sridhar ◽

Jenna Read ◽

...

Keyword(s):

T Cells ◽

Antiretroviral Therapy ◽

Hiv Infection ◽

Immune Activation ◽

Cd4 T Cells ◽

Acute Infection ◽

Interferon Response ◽

Suppressive Therapy ◽

Hiv Reservoir ◽

Latent Hiv

Abstract Background Persistent HIV infection of long-lived resting CD4 T cells, despite antiretroviral therapy (ART), remains a barrier to HIV cure. Women have a more robust type 1 interferon response during HIV infection relative to men, contributing to lower initial plasma viremia. As lower viremia during acute infection is associated with reduced frequency of latent HIV infection, we hypothesized that women on ART would have a lower frequency of latent HIV compared to men. Methods ART-suppressed, HIV seropositive women (n = 22) were matched 1:1 to 22 of 39 ART-suppressed men. We also compared the 22 women to all 39 men, adjusting for age and race as covariates. We measured the frequency of latent HIV using the quantitative viral outgrowth assay, the intact proviral DNA assay, and total HIV gag DNA. We also performed activation/exhaustion immunophenotyping on peripheral blood mononuclear cells and quantified interferon-stimulated gene (ISG) expression in CD4 T cells. Results We did not observe evident sex differences in the frequency of persistent HIV in resting CD4 T cells. Immunophenotyping and CD4 T-cell ISG expression analysis revealed marginal differences across the sexes. Conclusions Differences in HIV reservoir frequency and immune activation appear to be small across sexes during long-term suppressive therapy.

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In Situ Characterization of Human Lymphoid Tissue Immune Cells by Multispectral Confocal Imaging and Quantitative Image Analysis; Implications for HIV Reservoir Characterization

Frontiers in Immunology ◽

10.3389/fimmu.2021.683396 ◽

2021 ◽

Vol 12 ◽

Author(s):

Eirini Moysi ◽

Perla M. Del Rio Estrada ◽

Fernanda Torres-Ruiz ◽

Gustavo Reyes-Terán ◽

Richard A. Koup ◽

...

Keyword(s):

T Cells ◽

B Cells ◽

Immune Cells ◽

Cd4 T Cells ◽

Lymphoid Tissue ◽

Immune Cell ◽

Hiv Reservoir ◽

Human Lymphoid Tissue

CD4 T cells are key mediators of adaptive immune responses during infection and vaccination. Within secondary lymphoid organs, helper CD4 T cells, particularly those residing in germinal centers known as follicular helper T cells (Tfh), provide critical help to B-cells to promote their survival, isotype switching and selection of high affinity memory B-cells. On the other hand, the important role of Tfh cells for the maintenance of HIV reservoir is well documented. Thus, interrogating and better understanding the tissue specific micro-environment and immune subsets that contribute to optimal Tfh cell differentiation and function is important for designing successful prevention and cure strategies. Here, we describe the development and optimization of eight multispectral confocal microscopy immunofluorescence panels designed for in depth characterization and immune-profiling of relevant immune cells in formalin-fixed paraffin-embedded human lymphoid tissue samples. We provide a comprehensive library of antibodies to use for the characterization of CD4+ T-cells -including Tfh and regulatory T-cells- as well as CD8 T-cells, B-cells, macrophages and dendritic cells and discuss how the resulting multispectral confocal datasets can be quantitatively dissected using the HistoCytometry pipeline to collect information about relative frequencies and immune cell spatial distributions. Cells harboring actively transcribed virus are analyzed using an in-situ hybridization assay for the characterization of HIV mRNA positive cells in combination with additional protein markers (multispectral RNAscope). The application of this methodology to lymphoid tissues offers a means to interrogate multiple relevant immune cell targets simultaneously at increased resolution in a reproducible manner to guide CD4 T-cell studies in infection and vaccination.

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Blind Uneven Proliferation of CD4+ T cells During Primary Infection Generates the Majority of the HIV Reservoir

10.1101/2020.04.06.20053231 ◽

2020 ◽

Author(s):

Florencia A. T. Boshier ◽

Daniel B. Reeves ◽

Elizabeth R. Duke ◽

David A. Swan ◽

Martin Prlic ◽

...

Keyword(s):

T Cells ◽

Cd4 T Cells ◽

Cellular Proliferation ◽

Chromosomal Dna ◽

Hiv Replication ◽

Hiv Reservoir ◽

Viral Spread ◽

Hiv Dna ◽

Latently Infected ◽

Memory Cd4 T Cells

AbstractThe HIV reservoir is a population of 1-10 million anatomically dispersed, latently infected memory CD4+ T cells in which an HIV DNA molecule is quiescently integrated into human chromosomal DNA. When antiretroviral therapy (ART) is stopped and HIV replication initiates in one of these cells, systemic viral spread resumes, rekindling progression to AIDS. Therefore, HIV latency prevents cure. The HIV reservoir contains clones: identical HIV sequences that are integrated within identical human chromosomal DNA locations. The presence of these clones demonstrates that proliferation of CD4+ T cells sustains infection despite ART. The reservoir has a precise structure consisting of a small number of large clones and a large number of small clones. However, the mechanisms leading to this structure have not been identified. We developed a mathematical model that recapitulates the profound depletion and brisk recovery of CD4+ T cells, reservoir creation, and viral load trajectory during primary HIV infection. We extended the model to simulate stochastically individual HIV reservoir clones and identified that uneven proliferation among clones during recovery from CD4+ lymphopaenia is sufficient to explain the observed clonal reservoir distribution. We project that within one month of infection 75-95% of reservoir cells are generated from cellular proliferation rather than denovo viral infection. Recent detection of HIV infected clones during the first 5 weeks of infection support our model’s predictions.

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