Human Immunodeficiency Virus Type 1 Can Establish Latent Infection in Resting CD4<sup>+</sup>T Cells in the Absence of Activating Stimuli

Swiggard, William J.; Baytop, Clifford; Yu, Jianqing; Dai, Jihong; Li, Chuanzhao; Schretzenmair, Richard; Theodosopoulos, Ted; O’Doherty, Una

doi:10.1128/jvi.79.22.14179-14188.2005

Cited by 167 publications

(213 citation statements)

References 79 publications

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“…Several studies have demonstrated that HIV-1 can establish latency by direct infection of resting CD4 + T cells in vitro (40,67,68). This strategy does have some advantages because it avoids the activation of T cells and utilizes wild-type HIV-1.…”

Section: Discussionmentioning

confidence: 99%

Small-molecule screening using a human primary cell model of HIV latency identifies compounds that reverse latency without cellular activation

Yang¹,

Xing²,

Shan³

et al. 2009

J. Clin. Invest.

209

287

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The development of highly active antiretroviral therapy (HAART) to treat individuals infected with HIV-1 has dramatically improved patient outcomes, but HAART still fails to cure the infection. The latent viral reservoir in resting CD4 + T cells is a major barrier to virus eradication. Elimination of this reservoir requires reactivation of the latent virus. However, strategies for reactivating HIV-1 through nonspecific T cell activation have clinically unacceptable toxicities. We describe here the development of what we believe to be a novel in vitro model of HIV-1 latency that we used to search for compounds that can reverse latency. Human primary CD4 + T cells were transduced with the prosurvival molecule Bcl-2, and the resulting cells were shown to recapitulate the quiescent state of resting CD4 + T cells in vivo. Using this model system, we screened small-molecule libraries and identified a compound that reactivated latent HIV-1 without inducing global T cell activation, 5-hydroxynaphthalene-1,4-dione (5HN). Unlike previously described latency-reversing agents, 5HN activated latent HIV-1 through ROS and NF-κB without affecting nuclear factor of activated T cells (NFAT) and PKC, demonstrating that TCR pathways can be dissected and utilized to purge latent virus. Our study expands the number of classes of latency-reversing therapeutics and demonstrates the utility of this in vitro model for finding strategies to eradicate HIV-1 infection.

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Section: Discussionmentioning

confidence: 99%

Small-molecule screening using a human primary cell model of HIV latency identifies compounds that reverse latency without cellular activation

Yang¹,

Xing²,

Shan³

et al. 2009

J. Clin. Invest.

209

287

View full text Add to dashboard Cite

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“…Our previous data (1,38) and those of Vatakis et al (41) suggest that, despite this restriction, HIV can integrate directly into rCD4 cells, although this occurs with slower kinetics and to lower levels in resting cells than in activated cells. Given that dN increase reverse transcription in resting T cells (17), we tested the effect of dN addition on HIV integration and 2LTR formation in resting T cells.…”

Section: Resting Cd4mentioning

confidence: 99%

Addition of Deoxynucleosides Enhances Human Immunodeficiency Virus Type 1 Integration and 2LTR Formation in Resting CD4⁺T Cells

Plesa

Dai

Baytop

et al. 2007

J Virol

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Resting CD4 ؉ T cells restrict human immunodeficiency virus (HIV) infection at a step prior to integration. Despite this restriction, we showed previously that HIV integration occurs in resting CD4؉ T cells in vitro, albeit at lower levels than in activated CD4؉ T cells. Here we show that addition of deoxynucleosides enhanced integration and 2LTR formation in resting CD4؉ T cells but that the kinetics were still significantly delayed compared to those of activated T cells. Thus, deoxynucleoside addition partially overcomes the restriction to HIV infection in resting CD4 ؉ T cells. Resting CD4ϩ T cells (rCD4 cells) appear to be a major target of human immunodeficiency virus (HIV) and simian immunodeficiency virus infections in vivo, especially in the gut during acute infection (12,20,42,43,47,48), yet in vitro rCD4 cells resist HIV infection (8,34,35,45). Understanding this paradox is important to HIV pathogenesis. Early in vitro studies showed that reverse transcription and possibly other postentry steps are restricted in rCD4 cells (3,35,37,45,46). However, extended kinetic studies showed that reverse transcription is not completely blocked in resting T cells but rather occurs with slower kinetics than in activated cells (8,29,33,39). The slower kinetics of reverse transcription in resting T cells can be partially overcome by inducing the cells to enter G 1b (11,18,40), by knocking down putative restriction factors (8, 36) and by adding deoxynucleosides (dN) (17), which are present at lower levels in resting cells than in activated cells (9,10,24,26,32).Our previous data (1, 38) and those of Vatakis et al. (41) suggest that, despite this restriction, HIV can integrate directly into rCD4 cells, although this occurs with slower kinetics and to lower levels in resting cells than in activated cells. Given that dN increase reverse transcription in resting T cells (17), we tested the effect of dN addition on HIV integration and 2LTR formation in resting T cells. Here we show that dN increase integration and 2LTR formation in rCD4 cells.Two populations of CD4 ؉ T cells were prepared from PBMCs. To study the restriction of HIV infection by rCD4 cells, we prepared rCD4 cells and activated CD4 ϩ T cells (aCD4 cells) from peripheral blood mononuclear cells (PBMCs; Fig. 1A).Partially purified CD4 ϩ T cells (ppCD4 cells) were prepared by negative selection (Rosette Sep; Stem Cell Technologies) for other cell lineages (CD8, CD16, CD19, CD36, and CD56) (Fig. 1A). rCD4 cells were prepared from the ppCD4 cells by a second negative selection using phycoerythrin-labeled antibodies for ␥␦ T-cell receptor (␥␦TCR) and activation markers (CD69, CD25, and HLA-DR) followed by anti-phycoerythrin magnetic beads (magnetically activated cell sorting; Miltenyi). The resulting cells were 99% pure rCD4 cells (Fig. 1A). Preactivated CD4 ϩ T cells were prepared by stimulating ppCD4 cells with anti-CD3/28-coated beads (Dynal) for 3 days (5, 6). The aCD4 cells uniformly expressed CD25 and CD69 by day 3 (see Fig. 3).dN enhanced integration and 2LT...

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“…Subsequent studies, using more-sensitive techniques, established that RT in quiescent cells was far slower and less stable than in activated cells (17,36). In addition, we and others demonstrated that while cells activated for 48 h can be efficiently infected, infection prior to T-cell activation is very inefficient, characterized by a large delay in the kinetics of RT and integration, even if the cell is stimulated immediately following infection (11,18,27,30). Therefore, the activation state of T cells markedly affects the progression of the early stages of HIV infection (29).…”

Section: Unlike Activated T Cells Quiescent Cd4mentioning

confidence: 99%

Differentially Stimulated CD4⁺T Cells Display Altered Human Immunodeficiency Virus Infection Kinetics: Implications for the Efficacy of Antiviral Agents

Vatakis

Nixon²,

Bristol

et al. 2009

J Virol

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The activation state of CD4؉ T cells plays a crucial role in the establishment of a productive human immunodeficiency virus infection. Here, we show that T cells stimulated for 1 day demonstrated delayed kinetics of viral reverse transcription and integration compared to cells stimulated for 2 days prior to infection. As a result, the efficiency of reverse transcription and integration inhibitors differs in these differentially stimulated cells. These studies increase our understanding of how T cells support viral replication and provide insight regarding the efficiency of antiretroviral therapy in lymphoid compartments.

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Human Immunodeficiency Virus Type 1 Can Establish Latent Infection in Resting CD4⁺T Cells in the Absence of Activating Stimuli

Cited by 167 publications

References 79 publications

Small-molecule screening using a human primary cell model of HIV latency identifies compounds that reverse latency without cellular activation

Small-molecule screening using a human primary cell model of HIV latency identifies compounds that reverse latency without cellular activation

Addition of Deoxynucleosides Enhances Human Immunodeficiency Virus Type 1 Integration and 2LTR Formation in Resting CD4⁺T Cells

Differentially Stimulated CD4⁺T Cells Display Altered Human Immunodeficiency Virus Infection Kinetics: Implications for the Efficacy of Antiviral Agents

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Human Immunodeficiency Virus Type 1 Can Establish Latent Infection in Resting CD4+T Cells in the Absence of Activating Stimuli

Cited by 167 publications

References 79 publications

Small-molecule screening using a human primary cell model of HIV latency identifies compounds that reverse latency without cellular activation

Small-molecule screening using a human primary cell model of HIV latency identifies compounds that reverse latency without cellular activation

Addition of Deoxynucleosides Enhances Human Immunodeficiency Virus Type 1 Integration and 2LTR Formation in Resting CD4+T Cells

Differentially Stimulated CD4+T Cells Display Altered Human Immunodeficiency Virus Infection Kinetics: Implications for the Efficacy of Antiviral Agents

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Human Immunodeficiency Virus Type 1 Can Establish Latent Infection in Resting CD4⁺T Cells in the Absence of Activating Stimuli

Addition of Deoxynucleosides Enhances Human Immunodeficiency Virus Type 1 Integration and 2LTR Formation in Resting CD4⁺T Cells

Differentially Stimulated CD4⁺T Cells Display Altered Human Immunodeficiency Virus Infection Kinetics: Implications for the Efficacy of Antiviral Agents