- Open Access
Impact of LEDGIN treatment during virus production on residual HIV-1 transcription
© The Author(s) 2019
- Received: 20 February 2019
- Accepted: 23 March 2019
- Published: 2 April 2019
Persistence of latent, replication-competent provirus is the main impediment towards the cure of HIV infection. One of the critical questions concerning HIV latency is the role of integration site selection in HIV expression. Inhibition of the interaction between HIV integrase and its chromatin tethering cofactor LEDGF/p75 is known to reduce integration and to retarget residual provirus to regions resistant to reactivation. LEDGINs, small molecule inhibitors of the interaction between HIV integrase and LEDGF/p75, provide an interesting tool to study the underlying mechanisms. During early infection, LEDGINs block the interaction with LEDGF/p75 and allosterically inhibit the catalytic activity of IN (i.e. the early effect). When present during virus production, LEDGINs interfere with proper maturation due to enhanced IN oligomerization in the progeny virions (i.e. the late effect).
We studied the effect of LEDGINs present during virus production on the transcriptional state of the residual virus. Infection of cells with viruses produced in the presence of LEDGINs resulted in a residual reservoir that was refractory to activation. Integration of residual provirus was less favored near epigenetic markers associated with active transcription. However, integration near H3K36me3 and active genes, both targeted by LEDGF/p75, was not affected. Also in primary cells, LEDGIN treatment induced a reservoir resistant to activation due to a combined early and late effect.
LEDGINs present a research tool to study the link between integration and transcription, an essential question in retrovirology. LEDGIN treatment during virus production altered integration of residual provirus in a LEDGF/p75-independent manner, resulting in a reservoir that is refractory to activation.
- HIV latency
Potent combination antiretroviral therapy (cART) suppresses the plasma viral load of HIV infected patients to undetectable levels. As a result the quality of life has improved significantly and the number of AIDS-related deaths has drastically declined worldwide . Still, globally HIV remains the major cause of death among women between 15 and 49 years old . Moreover, it is challenging from a logistic and economic point of view to provide over 36 million HIV positive patients worldwide with a lifelong treatment [2–4]. In addition, drug related side effects hamper adherence to therapy and allow the emergence of drug resistant HIV strains [5–7]. Therefore, the development of new strategies towards a cure of HIV infection is crucial. The major barrier towards such cure is the persistence of integrated provirus residing mainly in long-lived latently infected memory CD4+ T cells responsible for a rebound in viremia upon treatment interruption [8–10]. Although no consensus exists about its relative importance, homeostatic proliferation and clonal expansion of infected cells contribute to viral persistence even under cART [11–17]. Low levels of ongoing viral replication in anatomical sanctuaries with limited drug penetration like the central nervous system, gut associated lymphoid tissue and the lymph nodes may maintain the reservoir as well [18, 19].
Integration of the viral DNA into the host genome is a crucial step in the formation of the proviral reservoir. It is catalyzed by HIV integrase (IN) (For a Review see ) with the help of Lens epithelium-derived growth factor (LEDGF/p75). LEDGF/p75 is a transcriptional co-activator that binds IN and tethers the pre-integration complex (PIC) to the chromatin [21–26], facilitating integration into transcriptionally active regions . LEDGF/p75 is not the only determinant of integration site selection . Nuclear import of HIV through nuclear pore complexes (NPC)  is a first important step that determines the path through which a PIC enters the nucleus [30, 31]. HIV-1 integration preferentially occurs in the nuclear periphery [32, 33] in active chromatin regions close to the nuclear pore, while inner nuclear or heterochromatic regions are apparently disfavored [34–36]. Depletion of several NPC associated proteins (Nup98, Nup153, Transportin-3, RanBP2 and Tpr) hampers integration in gene dense regions [36–38]. Cleavage and polyadenylation specificity factor 6 (CPSF6) is another cellular cofactor that promotes nuclear entry through its interaction with the HIV capsid protein [39–41]. Depletion or knockout of CPSF6 affects HIV integration in active genes [42–45]. Active genes are characterized by an open chromatin landscape and specific epigenetic histone modifications such as H3K36me3, the recognition mark of LEDGF/p75 [46–48]. Moreover, it was recently found that HIV IN directly interacts with the amino-terminal tail of histone H4, which promotes its anchoring to the nucleosome and facilitates integration . Additionally, HIV IN shows a weak preference for a conserved sequence logo at the site of integration [50–52].
To elaborate this hypothesis, we now investigated the late effect of LEDGINs on integration site selection and HIV-1 expression. In this study, we infected cells with HIV produced in the presence of LEDGINs and determined integration sites and reactivation potential. The fact that LEDGINs inhibit the late replication steps at a lower dose than the early steps [64–68], underscores the translational relevance of this question. LEDGIN treatment during virus production effectively altered immediate quiescence and the activation potential both in cell lines and primary CD4+ T cells. Although targeting to H3K36me3 and active genes, were unaffected, the residual integration sites were less favored near features associated with active transcription, reminiscent of a more latent chromatin landscape. These data suggest a LEDGF/p75-independent mechanism generating the silent phenotype observed after LEDGIN treatment during virus production.
LEDGIN treatment during virus production reduces infectivity and increases the proportion of quiescent provirus in cell lines
Next, we investigated whether LEDGIN treatment in producer cells influences HIV expression to a similar extent as previously documented for LEDGIN treatment during infection . Three days post infection, the quiescent fraction was determined as the ratio of single mKO2 positive cells over the total number of detected eGFP and mKO2 positive cells (C/(A + B + C) × 100, Fig. 2b). With increasing concentrations of CX014442 an augmentation in the quiescent fraction from about 70-80% up to 97% was observed in both Jurkat and SupT1 cells (Fig. 4c, f). In MT-4 cells, the quiescent fraction increased from less than 10% in the DMSO condition (Additional file 1: Table S1) to 50% with virus produced in the presence of 0.25 µM of CX014442 (Fig. 4i). The quiescent fraction calculated for the DMSO control for the different cell lines is shown in Additional file 1: Table S1. We conclude that LEDGIN treatment during virus production reduces infection of target cells in a dose-dependent manner and that residual integrants are more often in a transcriptionally silent state.
LEDGIN treatment during virus production results in a quiescent reservoir refractory to reactivation
The chromatin environment of residual provirus after LEDGIN treatment during virus production is associated with latency
LEDGIN treatment inhibits integration and HIV activation in primary CD4+ T cells
The latent HIV reservoir is the main target for different HIV cure strategies . The so-called shock-and-kill strategy aims to reactivate latent provirus with latency reversing agents (LRAs) and subsequent killing of the reactivated cells by viral cytopathic effects or by immune clearance [76–81]. So far results are hampered by insufficient potency and toxicity of latency reversing agents (LRA) [82–85]. It was shown that LRAs reactivate less than 5% of latent provirus  and that the response to different LRAs depends on the site of integration . This might explain the limited success of the shock-and-kill strategy. Therefore, it remains important to explore other cure strategies. Recently, a novel block-and-lock strategy was proposed that aims at locking the residual virus into a ‘deep’ latent or transcriptionally silent state lacking the capability to rebound upon cART cessation [57, 76, 86–92]. This deep latent state can be achieved in several ways. HIV transcription can be abrogated by inhibition of trans-activator of transcription (Tat) [86–88]. Alternatively, HIV expression and reactivation can be inhibited by LEDGIN-mediated retargeting of HIV integration to sites that are less susceptible to reactivation . To achieve an HIV cure via any of the discussed strategies, it is important to understand the role of integration site selection in HIV latency and reactivation. In this study, we further explored this relation by investigating the late effect of LEDGINs on residual HIV integration and expression.
LEDGINs were added during virus production (late effect), resulting in crippled progeny virions with an enhanced IN oligomerization [64–67]. First, we confirmed that LEDGIN treatment in producer cells reduces infectivity of the progeny virus in the next round of infection when using a dual colored reporter virus. These results are in full agreement with previously published data on the late effect of LEDGINs [64–67]. Next, we evaluated whether LEDGIN treatment during virus production alters HIV expression. After treatment with 0.25 µM LEDGIN CX014442 97% of residual provirus in Jurkat and SupT1 cells was in a quiescent state and less susceptible to reactivation. In the DMSO control only 70–80% of infected cells were quiescent. This result phenocopies the effect seen with LEDGIN treatment during infection . In MT-4 cells the quiescent fraction was also augmented with increasing concentration of CX014442. Whereas in the DMSO condition less than 10% of provirus was quiescent, treatment with 0.25 µM of LEDGIN increased the proportion of silent provirus up to 50%. Notably, the extent of eGFP expression in the DMSO control samples varied among cell lines due to different activation of the LTR promoter. This emphasizes the importance of comparing latency and reactivation phenotypes in multiple cell lines in parallel. The MT-4 cell line is transformed with human T cell lymphotropic virus type I (HTLV-1), activating the host T cell . Since MT-4 cells already constitute an active cell line, it is more difficult to achieve additional activation. In fact, we found that more than 90% of the MT-4 cells in the DMSO control were productively infected, and therefore MT-4 cells were not included in the reactivation experiments.
Next, we investigated whether retargeting of integration sites can explain the quiescent phenotype observed with viruses produced in the presence of LEDGINs. LEDGF/p75 is the main tethering factor guiding HIV to active transcription units  and depletion of LEDGF/p75 or disruption of its interaction with HIV IN is known to shift integration out of active genes [53–57]. Interestingly, CX014442 treatment during virus production did not significantly alter the percentage of integration in refSeq genes or near H3K36me3, the recognition mark of LEDGF/p75, in contrast to integration sites obtained with CX014442 treatment during early infection (Additional file 1: Table S2, data from ). The crippled viruses likely still depend on LEDGF/p75 for integration. When comparing integration sites after early or late LEDGIN treatment a similar pattern is observed (Additional file 1: Table S4). In both cases, integration becomes less favored near DNaseI, CpG islands and active transcription markers, while integration near transcriptionally silent markers is enriched upon addition of LEDGIN CX014442. Integration sites obtained after CX014442 treatment during virus production or during early infection of cells are both less favored in GC rich regions, although the effect is more pronounced when CX014442 is added during production. Overall, it seems that the main difference lies in targeting to refSeq genes. The IC50 value for the late effect of LEDGIN CX014442 is much lower, only up to 0.25 µM of compound was added during production. For the early effect, concentrations from 0.78 up to 50 µM were used (Additional file 1: Table S2). Higher concentrations of LEDGIN are thus required to shift integration out of refSeq genes. Changes in the chromatin environment on the other hand, already occur at the low LEDGIN concentrations used during production. Since these low concentrations are sufficient to cause the latent phenotype, the data suggest that a mechanism other than pure retargeting by LEDGF/p75 is involved. This is in agreement with our previous observation that the LEDGIN induced quiescent phenotype is stronger than the mere effect on retargeting . Enhanced IN oligomerization is a common feature seen both for early and late effect of LEDGINs and might play a role in this latent phenotype. Using Förster resonance energy transfer (FRET) we previously demonstrated a LEDGF/p75-dependent increase in IN multimerization in the nucleus . Upon knockdown of LEDGF/p75, this phenotype was rescued by addition of LEDGINs during infection of the cells . Moreover, LEDGIN treatment during virus production enhances IN oligomerization prematurely in the viral particle, and these multimers are retained in the cytoplasm and nucleus after infection of target cells [64, 94]. Although enhanced/premature IN oligomerization may not affect targeting by LEDGF/p75, it might still influence in which chromatin environment integration takes place, for instance by a direct interaction between IN and histone amino-terminal tails . The exact mechanism remains to be clarified.
Finally, our data obtained in primary CD4+ T cells validate the use of LEDGINs in a more clinically relevant acute infection model. LEDGIN CX014442 hampered WT HIV-1 activation in a dose-dependent manner. Although CX014442 was added during infection, the IC50 value obtained when using WT HIV-1 (237 ± 0.28 nM) was 26-fold lower compared to the IC50 value for single round infection of primary cells with the OGH virus (6.21 ± 0.22 µM) . This is due to the combination of the early and late effect of LEDGINs during multiple round replication with WT HIV-1. During multiple round infection in the presence of LEDGINs most likely a combination of LEDGF/p75-dependent and independent effects on integration site selection occur. How the observed effects possibly translate into patients and whether LEDGINs can be used in a block-and-lock functional cure needs to be investigated in advanced latency cell models, humanized mouse models and eventually in clinical trials.
Altogether, our data provide additional evidence for a link between integration site selection and HIV expression. LEDGIN treatment during virus production resulted in a residual reservoir that was more often in a quiescent state and refractory to activation. Integration was less favored in transcriptionally active chromatin, however, still mainly occurred in refSeq genes. In contrast to the quiescent phenotype seen upon LEDGIN treatment during early infection which is LEDGF/p75-dependent , we now observe a LEDGF/p75-independent phenotype. Possibly LEDGIN-enhanced IN oligomerization interferes with proper integration site selection. Our research shows that LEDGINs are a useful tool to investigate the importance of integration site selection and provide a rationale to further study their effects in context of a future block-and-lock cure strategy.
Cell culture and virus production
All cells were verified to be mycoplasma free by a cellular colorimetric detection assay (PlasmoTest™, InvivoGen Europe). Cells were cultured at 37 °C in a humidified atmosphere containing 5% CO2. SupT1 cells (provided by the National Institutes of Health reagent program, NIH, Bethesda, MD) [95, 96] were cultured in RPMI medium (GIBCO BRL) with 10% (v/v) fetal bovine serum (FBS, GIBCO) and 0.01% (v/v) gentamicin (GIBCO). HEK293T cells (generous gift from O. Danos, Evry, France) were cultured in Dulbecco Modified Eagle Medium (DMEM, GIBCO, Dublin, Ireland) with 5% (v/v) fetal bovine serum (FBS, GIBCO) and 0.01% v/v gentamicin (GIBCO). HEK293T cells were co-transfected, using linear polyethylenimine (PEI, Polysciences), with a plasmid encoding a single round HIV virus (pOGH) [70, 71] and a vesicular stomatitis virus G (VSV-G) protein encoding plasmid to produce VSV-G-pseudotyped viruses (Fig. 3a). Cells were washed twice with Phosphate Buffered Saline (PBS) to remove the excess of plasmid and the medium was replaced by medium containing different concentrations (7.8 nM–1 µM) of LEDGIN CX014442  6 h post transfection. The supernatant was collected 72 h post transfection and filtered through a 0.45 µm pore membrane (Merck, Overijse, Belgium). The virus was concentrated using a Vivaspin with a 15–50 kDa cut-off column (Merck) and washed three times with PBS. Next, the virus was treated with 100 U/ml DNase (Roche Diagnostics, Vilvoorde, Belgium) for 1 h at 37 °C to eliminate remaining plasmid and stored at − 80 °C. The vector used for integration site sequencing was produced by triple transfection with the transfer plasmid pCH-SFFV-eGFP-P2A-fLuc together with the Δ8.91 packaging plasmid and pVSV-G.
A multi-colored reporter virus (OGH) (Fig. 2a) was used to study the late effect of LEDGINs. This green-orange variant of the recently described LAI-based double reporter virus [70, 71] contains an LTR-driven enhanced Green Fluorescent Protein (eGFP) in the Nef gene position and a constitutively active EF1α promotor driving mutant Kusabira-Orange 2 (mKO2) expression . Simultaneous flow cytometry measurement of both reporters allows characterization of quiescent and active provirus (Fig. 2b) [97, 98]. Infected cells that are exclusively mKO2 positive, due to its expression driven by the internal constitutive promotor, are considered to comprise the LTR-silent or quiescent proviral pool. Cells expressing both mKO2 and eGFP are considered productively infected, as LTR-driven transcription is activated in these cells.
Reactivation experiments in cell lines
300,000 cells (Jurkat, SupT1 and MT-4 cells) were infected for 3 days in a 48-well plate with different dilutions of OGH virus that was produced in the presence of LEDGIN CX014442 as described above. The viral stocks were normalized on their p24 content (Innotest HIV antigen mAb, Fujirebio Europe). 72 h post infection, cells were washed twice with PBS and reseeded in a 12-well plate (Fig. 3b). At day eight, cells were reactivated in duplicate using 10 ng/ml Tumor Necrosis Factor α (TNFα, Immunosource, Zoersel, Belgium). Flow cytometry was performed on samples taken 3 days after infection and on day nine, 1 day after reactivation.
Flow cytometry analysis
Fluorescence was measured after cells were fixed in 2% paraformaldehyde (PFA) for 15 min at room temperature (RT) using a MACS Quant VYB analyzer (Miltenyi Biotech GmbH, Bergisch Gladbach, Germany). To measure eGFP expression, cells were excited using a 488 nm, 50 mW DPSS (diode-pumped solid-state) laser and the emitted signal passed through a 525/50 nm band pass filter. For mKO2 expression a 561 nm, 100 mW diode laser and 586/15 nm band pass filter were used. Live cells were selected based on the forward and side scatter channel (FSC-H/SSC-H) and doublets were excluded based on the FSC-A/FSC-H plot. For experiments in cell lines, at least 25,000 single live cells were counted in total and each sample was measured in duplicate. Single reporter constructs were used as controls. For flow cytometry analysis of primary CD4+ T cells infected with WT NL4.3-eGFP virus, at least 100,000 cells were counted. Data were analyzed using the FlowJo software (FlowJo LCC, Ashland, Oregon).
Integration site sequencing
Integration sites were determined as described previously . 100,000 SupT1 cells were transduced for 3 days with a lentiviral vector (CH-SFFV-eGFP-P2A-fLuc) that was produced in the presence of LEDGIN CX014442. Next, they were washed twice with PBS and kept in culture for at least 10 days to eliminate non-integrated DNA. Genomic DNA was extracted using the QIAamp DNA Mini kit (Qiagen). Genomic DNA was randomly sheared by sonication with the Covaris M220 and linkers were added to the sheared DNA ends. Integration sites were amplified by nested PCR using primers complementary to the linker and viral long terminal repeats (LTR). PCR products were sequenced by Illumina Miseq, paired-end 300 cycles. The INSPIIRED software  was used to analyze sequencing data.
Isolation of resting CD4+ T-cells
Human peripheral blood mononuclear cells (PBMC) were isolated from fresh buffy coats obtained from the Red Cross Blood transfusion Center (Mechelen, Belgium) using a lymphoprep density gradient centrifugation (Stem cell technologies, Cologne, Germany). Resting CD4+ T cells were enriched using a custom-made Easysep negative selection kit (Stem Cell Technologies, #19052 with the addition of CD25, CD69, and HLA-DR antibodies) and magnetic beads (Stem Cell Technologies), resulting in a purity of 95%. The experiments with human blood cells received bioethical approval by the Medical Ethics committee of the KU Leuven (S58969-IRB00002047).
Infection and activation of primary CD4+ T-cells
Freshly isolated resting CD4+ T cells were activated with 100 U/ml IL-2 (Peprotech, London, UK) and 10 µg/ml phytohaemagglutinin (PHA, Sigma) 48 h before infection. Cells were infected with wild type (WT) NL4.3 or WT NL4.3-eGFP virus (1.6 × 106 ng p24 per 1 × 107 cells/mL) for 3 h at 37 °C. Next, the excess of virus was washed away with PBS (three times) and cells were resuspended in RPMI medium with 10% (v/v) FBS and 0.1% gentamicin, supplemented with 1 U/ml IL-2 (Peprotech) and varying concentrations of LEDGIN CX014442 or raltegravir (provided by National Institute of Health AIDS reagent program, NIH, Bethesda, MD). Four days post infection compounds and virus were again washed away with PBS (three times). Cells were replated and activated with 10 nM phorbol myristate acetate (PMA, Sigma) and 10 µg/ml PHA or left untreated. Activation was quantified by measuring the viral p24 concentration in the culture supernatant 7 days post infection (Innotest HIV antigen mAb, Fujirebio Europe) and by flow cytometry for cells infected with WT NL4.3-eGFP virus. Four days post infection cells were harvested to determine the number of integrated copies using Alu-LTR qPCR and via flow cytometry for cells infected with WT NL4.3-eGFP.
Quantification of total integrated copy number
Integrated HIV DNA was quantified using a nested real-time Alu-LTR qPCR . 1 million cells were lysed in 50 µl of lysis buffer for 1 h at 56 °C (10 mM Tris HCl pH8, 1 mM EDTA, 0.01% triton and 0.8 mg/ml Proteinase K (PK)). The first round PCR reaction mix consisted of 5 µl of DNA from lysed cells, 12.5 µl of iQ supermix (Bio rad, Temse, Belgium), 0.5 µl of each primer (20 µM, Alu forward: TCCCAGCTACTGGGGAGGCTGAGG, Alu reverse: TGCTGGGATTACAGGCGTGAG and HIV-1 LTR forward: GCTAACTAGGGAACCCACTGCTTA) and 6 µl of water. Cycling conditions for the first round PCR were 95 °C for 10 min, followed by 15 cycles of 95 °C for 30 s, 60 °C for 40 s and 72 °C for 3.5 min. All samples were run at least in duplicate. 5 µl of the first-round product was added to a second round PCR mix containing 12.5 µl of iQ supermix, 0.5 µl of forward and reverse primer (20 µM, HIV-1 LTR forward: AGCTTGCCTTGAGTGCTTCAA, HIV-1 LTR reverse: TGACTAAAAGGGTCTGAGGGATCT), 1 µl of probe (5 µM, 5′-FAM-TTACCAGAGTCACACAACAGACGGGCA-TAMRA-3′) and 5.5 µl of water. Second round PCR was performed in a LightCycler 480 (Roche Life Science, Vilvoorde, Belgium) for 5 min at 95 °C, followed by 45 cycles of 95 °C for 15 s, 60 °C for 30 s and 72 °C for 1 min. As a standard genomic DNA of SupT1 cells transduced with the OGH virus and passaged for 3 weeks, was used. Integrated copies were normalized for input DNA by a parallel CCR5 qPCR as previously described . Data were analyzed using the provided LightCycler 480 software.
All data was analysed using the GraphPad Prism software version 7.00 for Windows (GraphPad Software, La Jolla California USA). IC50 values were calculated via a nonlinear regression curve fit of the concentration of inhibitor versus response. The statistical significance of the effect of LEDGINs compared to the control sample was assessed via one-way ANOVA with Dunnett’s multiple comparisons test in cell lines or with the Kruskal–Wallis test for primary cell data.
ZD conceived the study. GV, LV, IZ, DVL, PV, RG, FC and ZD designed experiments. GV, IZ and PV conducted experiments. GV, LV, IZ, DVL, PV, RG, FC and ZD analyzed the data. GV and CN performed bioinformatic analysis. GV and ZD prepared the manuscript. All authors read and approved the final manuscript.
We are grateful to B. Van Remoortel (Molecular Virology and Gene Therapy, KU Leuven) for the technical assistance and to K. Jacobs (Center for cancer biology, VIB-KU Leuven) for helping with flow cytometry measurements. Illumina sequencing was performed by Genomics Core Leuven. LEDGINs were synthesized by Cistim/CD3 (courtesy of Dr. A. Marchand). The double reporter virus was a kind gift from the Verdin lab (Buck Institute for Research on Aging).
The authors declare that they have no competing interests.
Availability of data and materials
Integration site sequencing data obtained in this study are deposited to the Sequence Read Archive. Integration sites for viruses produced in the presence of LEDGIN CX014442, SRA accession SRP157991 is available via following link: https://www.ncbi.nlm.nih.gov/sra/SRP157991.
Consent for publication
Ethics approval and consent to participate
The experiments with human blood cells received bioethical approval by the Medical Ethics committee of the KU Leuven (S58969-IRB00002047).
This work was supported by the Flemish Fund for Scientific Research (FWO; Fonds voor Wetenschappelijk Onderzoek) to [GV]. Research at KU Leuven received financial support from the FWO, the KU Leuven Research Council [OT; OT/13/098], HIV-ERA EURECA [IWT-SBO-EURECA, ZL345530], the KU Leuven Interdisciplinary Research (IDO) program [IDO/12/008], the Belgian IAP Belvir [ZKC4893-P7/45-P]. The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
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- Unaids. UNAIDS Data 2017. Program HIV/AIDS. 2017;1–248. http://www.unaids.org/sites/default/files/media_asset/20170720_Data_book_2017_en.pdf. Accessed 21 June 2018.
- UNAIDS (Joint United Nations Programme on HIV/AIDS). Access to antiretroviral therapy in Africa, 2013. Status Rep Prog Towar 2015 targets. 2013;1–10. www.unaids.org. Accessed 1 Jul 2018.
- Freedberg KA, Losina E, Weinstein MC, Paltiel AD, Cohen CJ, Seage GR, et al. The cost effectiveness of combination antiretroviral therapy for HIV disease. N Engl J Med. 2001;344(11):824–31. https://doi.org/10.1056/NEJM200103153441108.PubMedView ArticleGoogle Scholar
- UNAIDS. Facts sheet November 2016. http://www.unaids.org/. Accessed 1 Jul 2018.
- Meresse M, March L, Kouanfack C, Bonono R-C, Boyer S, Laborde-Balen G, et al. Patterns of adherence to antiretroviral therapy and HIV drug resistance over time in the Stratall ANRS 12110/ESTHER trial in Cameroon. HIV Med. 2014;15(8):478–87. https://doi.org/10.1111/hiv.12140.PubMedView ArticleGoogle Scholar
- Harrigan PR, Hogg RS, Dong WWY, Yip B, Wynhoven B, Woodward J, et al. Predictors of HIV drug-resistance mutations in a large antiretroviral-naive cohort initiating triple antiretroviral therapy. J Infect Dis. 2005;191(3):339–47. https://doi.org/10.1086/427192.PubMedView ArticleGoogle Scholar
- O’Brien ME, Clark RA, Besch CL, Myers L, Kissinger P. Patterns and correlates of discontinuation of the initial HAART regimen in an urban outpatient cohort. J Acquir Immune Defic Syndr. 2003;34(4):407–14.PubMedView ArticleGoogle Scholar
- Finzi D, Hermankova M, Pierson T, Carruth LM, Buck C, Chaisson RE, et al. Identification of a reservoir for HIV-1 in patients on highly active antiretroviral therapy. Science. 1997;278(5341):1295–300.PubMedView ArticleGoogle Scholar
- Chun T-W, Carruth L, Finzi D, Shen X, DiGiuseppe JA, Taylor H, et al. Quantification of latent tissue reservoirs and total body viral load in HIV-1 infection. Nature. 1997;387(6629):183–8. https://doi.org/10.1038/387183a0.PubMedView ArticleGoogle Scholar
- Chun T-W, Stuyver L, Mizell SB, Ehler LA, Mican JAM, Baseler M, et al. Presence of an inducible HIV-1 latent reservoir during highly active antiretroviral therapy. Proc Natl Acad Sci USA. 1997;94(24):13193–7.PubMedView ArticleGoogle Scholar
- Wagner TA, McLaughlin S, Garg K, Cheung CYK, Larsen BB, Styrchak S, et al. Proliferation of cells with HIV integrated into cancer genes contributes to persistent infection. Science. 2014;345(6196):570–3.PubMedPubMed CentralView ArticleGoogle Scholar
- Maldarelli F, Wu X, Su L, Simonetti FR, Shao W, Hill S, et al. Specific HIV integration sites are linked to clonal expansion and persistence of infected cells. Science. 2014;345(6193):179–83.PubMedPubMed CentralView ArticleGoogle Scholar
- Cohn LB, Silva IT, Oliveira TY, Rosales RA, Parrish EH, Learn GH, et al. HIV-1 integration landscape during latent and active infection. Cell. 2015;160(3):420–32.PubMedPubMed CentralView ArticleGoogle Scholar
- Murray AJ, Kwon KJ, Farber DL, Siliciano RF. The latent reservoir for HIV-1: how immunologic memory and clonal expansion contribute to HIV-1 persistence. J Immunol. 2016;197(2):407–17.PubMedPubMed CentralView ArticleGoogle Scholar
- Lee GQ, Orlova-Fink N, Einkauf K, Chowdhury FZ, Sun X, Harrington S, et al. Clonal expansion of genome-intact HIV-1 in functionally polarized Th1 CD4(+) T cells. J Clin Investig. 2017;127(7):2689–96.PubMedView ArticleGoogle Scholar
- Chun T-W, Stuyver L, Mizell SB, Ehler LA, Mican JAM, Baseler M, et al. Presence of an inducible HIV-1 latent reservoir during highly active antiretroviral therapy. Proc Natl Acad Sci. 1997;94(24):13193–7.PubMedView ArticleGoogle Scholar
- Hosmane NN, Kwon KJ, Bruner KM, Capoferri AA, Beg S, Rosenbloom DIS, et al. Proliferation of latently infected CD4+ T cells carrying replication-competent HIV-1: potential role in latent reservoir dynamics. J Exp Med. 2017;214(4):959–72. https://doi.org/10.1084/jem.20170193.PubMedPubMed CentralView ArticleGoogle Scholar
- Lorenzo-Redondo R, Fryer HR, Bedford T, Kim E-Y, Archer J, Kosakovsky Pond SL, et al. Persistent HIV-1 replication maintains the tissue reservoir during therapy. Nature. 2016;530(7588):51–6. https://doi.org/10.1038/nature16933.PubMedPubMed CentralView ArticleGoogle Scholar
- Fletcher CV, Staskus K, Wietgrefe SW, Rothenberger M, Reilly C, Chipman JG, et al. Persistent HIV-1 replication is associated with lower antiretroviral drug concentrations in lymphatic tissues. Proc Natl Acad Sci USA. 2014;111(6):2307–12.PubMedView ArticleGoogle Scholar
- Lesbats P, Engelman AN, Cherepanov P. Retroviral DNA integration. Chem Rev. 2016;116(20):12730–57.PubMedPubMed CentralView ArticleGoogle Scholar
- Debyser Z, Christ F, De Rijck J, Gijsbers R. Host factors for retroviral integration site selection. Trends Biochem Sci. 2015;40(2):108–16.PubMedView ArticleGoogle Scholar
- Cherepanov P, Maertens G, Proost P, Devreese B, Van Beeumen J, Engelborghs Y, et al. HIV-1 integrase forms stable tetramers and associates with LEDGF/p75 protein in human cells. J Biol Chem. 2003;278(1):372–81.PubMedView ArticleGoogle Scholar
- Llano M, Saenz DT, Meehan A, Wongthida P, Peretz M, Walker WH, et al. An essential role for LEDGF/p75 in HIV integration. Science. 2006;314(5798):461–4.PubMedView ArticleGoogle Scholar
- Maertens G, Cherepanov P, Pluymers W, Busschots K, De Clercq E, Debyser Z, et al. LEDGF/p75 is essential for nuclear and chromosomal targeting of HIV-1 integrase in human cells. J Biol Chem. 2003;278(35):33528–39.PubMedView ArticleGoogle Scholar
- McNeely M, Hendrix J, Busschots K, Boons E, Deleersnijder A, Gerard M, et al. In vitro DNA tethering of HIV-1 integrase by the transcriptional coactivator LEDGF/p75. J Mol Biol. 2011;410(5):811–30.PubMedView ArticleGoogle Scholar
- Vandekerckhove L, Christ F, Van Maele B, De Rijck J, Gijsbers R, Van den Haute C, et al. Transient and stable knockdown of the integrase cofactor LEDGF/p75 reveals its role in the replication cycle of human immunodeficiency virus. J Virol. 2006;80(4):1886–96.PubMedPubMed CentralView ArticleGoogle Scholar
- Schröder ARW, Shinn P, Chen H, Berry C, Ecker JR, Bushman F. HIV-1 integration in the human genome favors active genes and local hotspots. Cell. 2002;110(4):521–9.PubMedView ArticleGoogle Scholar
- Demeulemeester J, De Rijck J, Gijsbers R, Debyser Z. Retroviral integration: site matters: mechanisms and consequences of retroviral integration site selection. BioEssays. 2015;37(11):1202–14.PubMedPubMed CentralView ArticleGoogle Scholar
- Bukrinsky MI, Sharova N, Dempsey MP, Stanwick TL, Bukrinskaya AG, Haggerty S, et al. Active nuclear import of human immunodeficiency virus type 1 preintegration complexes. Proc Natl Acad Sci USA. 1992;89(14):6580–4.PubMedView ArticleGoogle Scholar
- Wong RW, Mamede JI, Hope TJ. Impact of nucleoporin-mediated chromatin localization and nuclear architecture on HIV integration site selection. J Virol. 2015;89(19):9702–5.PubMedPubMed CentralView ArticleGoogle Scholar
- Lusic M, Siliciano RF. Nuclear landscape of HIV-1 infection and integration. Nat Rev Microbiol. 2016;15:69. https://doi.org/10.1038/nrmicro.2016.162.PubMedView ArticleGoogle Scholar
- Burdick RC, Hu W-S, Pathak VK. Nuclear import of APOBEC3F-labeled HIV-1 preintegration complexes. Proc Natl Acad Sci. 2013;110(49):E4780–9.PubMedView ArticleGoogle Scholar
- Di Primio C, Quercioli V, Allouch A, Gijsbers R, Christ F, Debyser Z, et al. Single-cell imaging of HIV-1 provirus (SCIP). Proc Natl Acad Sci USA. 2013;110(14):5636–41.PubMedView ArticleGoogle Scholar
- Marini B, Kertesz-Farkas A, Ali H, Lucic B, Lisek K, Manganaro L. Nuclear architecture dictates HIV-1 integration site selection. Nature. 2015. https://doi.org/10.1038/nature14226.PubMedView ArticleGoogle Scholar
- Albanese A, Arosio D, Terreni M, Cereseto A. HIV-1 pre-integration complexes selectively target decondensed chromatin in the nuclear periphery. PLoS ONE. 2008;3(6):e2413. https://doi.org/10.1371/journal.pone.0002413.PubMedPubMed CentralView ArticleGoogle Scholar
- Lelek M, Casartelli N, Pellin D, Rizzi E, Souque P, Severgnini M, et al. Chromatin organization at the nuclear pore favours HIV replication. Nat Commun. 2015;6:6483. https://doi.org/10.1038/ncomms7483.PubMedPubMed CentralView ArticleGoogle Scholar
- Ocwieja KE, Brady TL, Ronen K, Huegel A, Roth SL, Schaller T, et al. HIV integration targeting: a pathway involving Transportin-3 and the nuclear pore protein RanBP2. PLoS Pathog. 2011;7(3):e1001313.PubMedPubMed CentralView ArticleGoogle Scholar
- Di Nunzio F, Fricke T, Miccio A, Valle-Casuso JC, Perez P, Souque P, et al. Nup153 and Nup98 bind the HIV-1 core and contribute to the early steps of HIV-1 replication. Virology. 2013;440(1):8–18.PubMedView ArticleGoogle Scholar
- Price AJ, Fletcher AJ, Schaller T, Elliott T, Lee K, KewalRamani VN, et al. CPSF6 defines a conserved capsid interface that modulates HIV-1 replication. PLoS Pathog. 2012;8(8):e1002896. https://doi.org/10.1371/journal.ppat.1002896.PubMedPubMed CentralView ArticleGoogle Scholar
- Matreyek KA, Yücel SS, Li X, Engelman A. Nucleoporin NUP153 phenylalanine-glycine motifs engage a common binding pocket within the HIV-1 capsid protein to mediate lentiviral infectivity. PLoS Pathog. 2013;9(10):e1003693.PubMedPubMed CentralView ArticleGoogle Scholar
- Bhattacharya A, Alam SL, Fricke T, Zadrozny K, Sedzicki J, Taylor AB, et al. Structural basis of HIV-1 capsid recognition by PF74 and CPSF6. Proc Natl Acad Sci USA. 2014;111(52):18625–30.PubMedView ArticleGoogle Scholar
- Chin CR, Perreira JM, Savidis G, Portmann JM, Aker AM, Feeley EM, et al. Direct visualization of HIV-1 replication intermediates shows that capsid and CPSF6 modulate HIV-1 intra-nuclear invasion and integration. Cell Rep. 2015;13(8):1717–31.PubMedPubMed CentralView ArticleGoogle Scholar
- Rasheedi S, Shun M-C, Serrao E, Sowd GA, Qian J, Hao C, et al. The cleavage and polyadenylation specificity factor 6 (CPSF6) subunit of the capsid-recruited pre-messenger RNA cleavage factor I (CFIm) complex mediates HIV-1 integration into genes. J Biol Chem. 2016;291(22):11809–19.PubMedPubMed CentralView ArticleGoogle Scholar
- Sowd GA, Serrao E, Wang H, Wang W, Fadel HJ, Poeschla EM, et al. A critical role for alternative polyadenylation factor CPSF6 in targeting HIV-1 integration to transcriptionally active chromatin. Proc Natl Acad Sci USA. 2016;113(8):E1054–63.PubMedView ArticleGoogle Scholar
- Achuthan V, Perreira JM, Sowd GA, Puray-Chavez M, McDougall WM, Paulucci-Holthauzen A, et al. Capsid-CPSF6 interaction licenses nuclear HIV-1 trafficking to sites of viral DNA integration. Cell Host Microbe. 2018;24(3):392–404.e8. https://doi.org/10.1016/j.chom.2018.08.002.PubMedView ArticleGoogle Scholar
- Eidahl JO, Crowe BL, North JA, McKee CJ, Shkriabai N, Feng L, et al. Structural basis for high-affinity binding of LEDGF PWWP to mononucleosomes. Nucleic Acids Res. 2013;41(6):3924–36.PubMedPubMed CentralView ArticleGoogle Scholar
- Pradeepa MM, Sutherland HG, Ule J, Grimes GR, Bickmore WA. Psip1/Ledgf p52 binds methylated histone H3K36 and splicing factors and contributes to the regulation of alternative splicing. PLoS Genet. 2012;8(5):e1002717.PubMedPubMed CentralView ArticleGoogle Scholar
- van Nuland R, van Schaik FM, Simonis M, van Heesch S, Cuppen E, Boelens R, et al. Nucleosomal DNA binding drives the recognition of H3K36-methylated nucleosomes by the PSIP1-PWWP domain. Epigenetics Chromatin. 2013;6(1):12.PubMedPubMed CentralView ArticleGoogle Scholar
- Benleulmi MS, Matysiak J, Robert X, Miskey C, Mauro E, Lapaillerie D, et al. Modulation of the functional association between the HIV-1 intasome and the nucleosome by histone amino-terminal tails. Retrovirology. 2017;14:54.PubMedPubMed CentralView ArticleGoogle Scholar
- Holman AG, Coffin JM. Symmetrical base preferences surrounding HIV-1, avian sarcoma/leukosis virus, and murine leukemia virus integration sites. Proc Natl Acad Sci USA. 2005;102(17):6103–7.PubMedView ArticleGoogle Scholar
- Wu X, Li Y, Crise B, Burgess SM, Munroe DJ. Weak palindromic consensus sequences are a common feature found at the integration target sites of many retroviruses. J Virol. 2005;79(8):5211–4.PubMedPubMed CentralView ArticleGoogle Scholar
- Berry C, Hannenhalli S, Leipzig J, Bushman FD. Selection of target sites for mobile DNA integration in the human genome. PLoS Comput Biol. 2006;2(11):e157. https://doi.org/10.1371/journal.pcbi.0020157.PubMedPubMed CentralView ArticleGoogle Scholar
- Ciuffi A, Llano M, Poeschla E, Hoffmann C, Leipzig J, Shinn P, et al. A role for LEDGF/p75 in targeting HIV DNA integration. Nat Med. 2005;11:1287. https://doi.org/10.1038/nm1329.PubMedView ArticleGoogle Scholar
- Shun M-C, Raghavendra NK, Vandegraaff N, Daigle JE, Hughes S, Kellam P, et al. LEDGF/p75 functions downstream from preintegration complex formation to effect gene-specific HIV-1 integration. Genes Dev. 2007;21(14):1767–78.PubMedPubMed CentralView ArticleGoogle Scholar
- Schrijvers R, De Rijck J, Demeulemeester J, Adachi N, Vets S, Ronen K, et al. LEDGF/p75-independent HIV-1 replication demonstrates a role for HRP-2 and remains sensitive to inhibition by LEDGINs. PLoS Pathog. 2012;8(3):e1002558.PubMedPubMed CentralView ArticleGoogle Scholar
- Fadel HJ, Morrison JH, Saenz DT, Fuchs JR, Kvaratskhelia M, Ekker SC, et al. TALEN knockout of the PSIP1 gene in human cells: analyses of HIV-1 replication and allosteric integrase inhibitor mechanism. J Virol. 2014;88(17):9704–17.PubMedPubMed CentralView ArticleGoogle Scholar
- Vranckx LS, Demeulemeester J, Saleh S, Boll A, Vansant G, Schrijvers R, et al. LEDGIN-mediated inhibition of integrase–LEDGF/p75 interaction reduces reactivation of residual latent HIV. EBioMedicine. 2016;8:248–64.PubMedPubMed CentralView ArticleGoogle Scholar
- Christ F, Voet A, Marchand A, Nicolet S, Desimmie BA, Marchand D, et al. Rational design of small-molecule inhibitors of the LEDGF/p75-integrase interaction and HIV replication. Nat Chem Biol. 2010;6(6):442–8. https://doi.org/10.1038/nchembio.370.PubMedView ArticleGoogle Scholar
- Al-Mawsawi LQ, Neamati N. Allosteric inhibitor development targeting HIV-1 integrase. ChemMedChem. 2011;6(2):228–41.PubMedPubMed CentralView ArticleGoogle Scholar
- Kessl JJ, Jena N, Koh Y, Taskent-Sezgin H, Slaughter A, Feng L, et al. Multimode, cooperative mechanism of action of allosteric HIV-1 integrase inhibitors. J Biol Chem. 2012;287(20):16801–11.PubMedPubMed CentralView ArticleGoogle Scholar
- Tsiang M, Jones GS, Niedziela-Majka A, Kan E, Lansdon EB, Huang W, et al. New class of HIV-1 integrase (IN) inhibitors with a dual mode of action. J Biol Chem. 2012;287(25):21189–203.PubMedPubMed CentralView ArticleGoogle Scholar
- Demeulemeester J, Chaltin P, Marchand A, De Maeyer M, Debyser Z, Christ F. LEDGINs, non-catalytic site inhibitors of HIV-1 integrase: a patent review (2006–2014). Expert Opin Ther Pat. 2014;24(6):609–32. https://doi.org/10.1517/13543776.2014.898753.PubMedView ArticleGoogle Scholar
- Christ F, Shaw S, Demeulemeester J, Desimmie BA, Marchand A, Butler S, et al. Small-molecule inhibitors of the LEDGF/p75 binding site of integrase block HIV replication and modulate integrase multimerization. Antimicrob Agents Chemother. 2012;56(8):4365–74.PubMedPubMed CentralView ArticleGoogle Scholar
- Desimmie BA, Schrijvers R, Demeulemeester J, Borrenberghs D, Weydert C, Thys W, et al. LEDGINs inhibit late stage HIV-1 replication by modulating integrase multimerization in the virions. Retrovirology. 2013;10:57.PubMedPubMed CentralView ArticleGoogle Scholar
- Jurado KA, Wang H, Slaughter A, Feng L, Kessl JJ, Koh Y, et al. Allosteric integrase inhibitor potency is determined through the inhibition of HIV-1 particle maturation. Proc Natl Acad Sci USA. 2013;110(21):8690–5.PubMedView ArticleGoogle Scholar
- Le Rouzic E, Bonnard D, Chasset S, Bruneau J-M, Chevreuil F, Le Strat F, et al. Dual inhibition of HIV-1 replication by integrase-LEDGF allosteric inhibitors is predominant at the post-integration stage. Retrovirology. 2013;10(1):144. https://doi.org/10.1186/1742-4690-10-144.PubMedPubMed CentralView ArticleGoogle Scholar
- Balakrishnan M, Yant SR, Tsai L, O’Sullivan C, Bam RA, Tsai A, et al. Non-catalytic site HIV-1 integrase inhibitors disrupt core maturation and induce a reverse transcription block in target cells. PLoS ONE. 2013;8(9):e74163. https://doi.org/10.1371/journal.pone.0074163.PubMedPubMed CentralView ArticleGoogle Scholar
- Bonnard D, Le Rouzic E, Eiler S, Amadori C, Orlov I, Bruneau J-M, et al. Structure-function analyses unravel distinct effects of allosteric inhibitors of HIV-1 integrase on viral maturation and integration. J Biol Chem. 2018;293(16):6172–86.PubMedView ArticleGoogle Scholar
- Chen H-C, Martinez JP, Zorita E, Meyerhans A, Filion GJ. Position effects influence HIV latency reversal. Nat Struct Mol Biol. 2017;24(1):47–54. https://doi.org/10.1038/nsmb.3328.PubMedView ArticleGoogle Scholar
- Battivelli E, Dahabieh MS, Abdel-Mohsen M, Svensson JP, Tojal Da Silva I, Cohn LB, et al. Distinct chromatin functional states correlate with HIV latency reactivation in infected primary CD4+ T cells. Elife. 2018;7:e34655. https://doi.org/10.7554/eLife.34655.001 PubMedPubMed CentralView ArticleGoogle Scholar
- Chavez L, Calvanese V, Verdin E. HIV latency is established directly and early in both resting and activated primary CD4 T cells. PLoS Pathog. 2015;11(6):e1004955.PubMedPubMed CentralView ArticleGoogle Scholar
- Berry CC, Nobles C, Six E, Wu Y, Malani N, Sherman E, et al. INSPIIRED: quantification and visualization tools for analyzing integration site distributions. Mol Ther Methods Clin Dev. 2017;4:17–26.PubMedView ArticleGoogle Scholar
- Mitchell RS, Beitzel BF, Schroder ARW, Shinn P, Chen H, Berry CC, et al. Retroviral DNA integration: ASLV, HIV, and MLV show distinct target site preferences. PLoS Biol. 2004;2(8):e234.PubMedPubMed CentralView ArticleGoogle Scholar
- Lewinski MK, Bisgrove D, Shinn P, Chen H, Hoffmann C, Hannenhalli S, et al. Genome-wide analysis of chromosomal features repressing human immunodeficiency virus transcription. J Virol. 2005;79(11):6610–9.PubMedPubMed CentralView ArticleGoogle Scholar
- Bushman F, Lewinski M, Ciuffi A, Barr S, Leipzig J, Hannenhalli S, et al. Genome-wide analysis of retroviral DNA integration. Nat Rev Microbiol. 2005;3:848. https://doi.org/10.1038/nrmicro1263.PubMedView ArticleGoogle Scholar
- Darcis G, Van Driessche B, Van Lint C. HIV latency: should we shock or lock? Trends Immunol. 2017;38(3):217–28.PubMedView ArticleGoogle Scholar
- Pham HT, Mesplède T. The latest evidence for possible HIV-1 curative strategies. Drugs Context. 2018;7:212522.PubMedPubMed CentralView ArticleGoogle Scholar
- Schwartz C, Bouchat S, Marban C, Gautier V, Van Lint C, Rohr O, et al. On the way to find a cure: purging latent HIV-1 reservoirs. Biochem Pharmacol. 2017;146:10–22.PubMedView ArticleGoogle Scholar
- Kim Y, Anderson JL, Lewin SR. Getting the “kill” into “shock and kill”: strategies to eliminate latent HIV. Cell Host Microbe. 2018;23(1):14–26.PubMedPubMed CentralView ArticleGoogle Scholar
- Dahabieh M, Battivelli E, Verdin E. Understanding HIV latency: the road to an HIV cure. Annu Rev Med. 2015;66:407–21.PubMedPubMed CentralView ArticleGoogle Scholar
- Xing S, Siliciano RF. Targeting HIV latency: pharmacologic strategies toward eradication. Drug Discov Today. 2013;18(11–12):541–51.PubMedView ArticleGoogle Scholar
- Gutiérrez C, Serrano-Villar S, Madrid-Elena N, Pérez-Elías MJ, Martín ME, Barbas C, et al. Bryostatin-1 for latent virus reactivation in HIV-infected patients on antiretroviral therapy. AIDS. 2016;30(9):1385–92.PubMedView ArticleGoogle Scholar
- Ho YC, Shan L, Hosmane NN, Wang J, Laskey SB, Rosenbloom DI. Replication-competent noninduced proviruses in the latent reservoir increase barrier to HIV-1 cure. Cell. 2013. https://doi.org/10.1016/j.cell.2013.09.020.PubMedPubMed CentralView ArticleGoogle Scholar
- Bullen CK, Laird GM, Durand CM, Siliciano JD, Siliciano RF. Novel ex vivo approaches distinguish effective and ineffective single agents for reversing HIV-1 latency in vivo. Nat Med. 2014;20(4):425–9.PubMedPubMed CentralView ArticleGoogle Scholar
- Zhao M, De Crignis E, Rokx C, Verbon A, van Gelder T, Mahmoudi T, et al. T cell toxicity of HIV latency reversing agents. Pharmacol Res. 2019;139:524–34. https://doi.org/10.1016/j.phrs.2018.10.023.PubMedView ArticleGoogle Scholar
- Mousseau G, Mediouni S, Valente ST. Targeting HIV transcription: the quest for a functional cure. Curr Top Microbiol Immunol. 2015;389:121–45.PubMedPubMed CentralGoogle Scholar
- Mousseau G, Kessing CF, Fromentin R, Trautmann L, Chomont N, Valente ST. The Tat inhibitor didehydro-cortistatin A prevents HIV-1 reactivation from latency. MBio. 2015;6(4):e00465-15.PubMedPubMed CentralView ArticleGoogle Scholar
- Kessing CF, Nixon CC, Li C, Tsai PM, Takata H, Mousseau G, et al. In vivo suppression of HIV rebound by didehydro-cortistatin A, a “block-and-lock” strategy for HIV-1 cure. Cell Rep. 2017;21(3):600–11.PubMedPubMed CentralView ArticleGoogle Scholar
- Jean MJ, Hayashi T, Huang H, Brennan J, Simpson S, Purmal A, et al. Curaxin CBL0100 blocks HIV-1 replication and reactivation through inhibition of viral transcriptional elongation. Front Microbiol. 2017;8:2007.PubMedPubMed CentralView ArticleGoogle Scholar
- Kim H, Choi M-S, Inn K-S, Kim B-J. Inhibition of HIV-1 reactivation by a telomerase-derived peptide in a HSP90-dependent manner. Sci Rep. 2016;6:28896.PubMedPubMed CentralView ArticleGoogle Scholar
- Anderson I, Low JS, Weston S, Weinberger M, Zhyvoloup A, Labokha AA, et al. Heat shock protein 90 controls HIV-1 reactivation from latency. Proc Natl Acad Sci USA. 2014;111(15):E1528–37.PubMedView ArticleGoogle Scholar
- Besnard E, Hakre S, Kampmann M, Lim HW, Hosmane NN, Martin A, et al. The mTOR complex controls HIV Latency. Cell Host Microbe. 2016;20(6):785–97.PubMedPubMed CentralView ArticleGoogle Scholar
- Höllsberg P. Mechanisms of T-cell activation by human T-cell lymphotropic virus type I. Microbiol Mol Biol Rev. 1999;63(2):308–33.PubMedPubMed CentralGoogle Scholar
- Borrenberghs D, Dirix L, De Wit F, Rocha S, Blokken J, De Houwer S, et al. Dynamic oligomerization of integrase orchestrates HIV nuclear entry. Sci Rep. 2016;6:36485. https://doi.org/10.1038/srep36485.PubMedPubMed CentralView ArticleGoogle Scholar
- Smith SD, Shatsky M, Cohen PS, Wamke R, Link MP, Glader BE. Monoclonal antibody and enzymatic profiles of human malignant T-lymphoid cells and derived cell lines. Cancer Res. 1984;44(12 Part 1):5657–60.PubMedGoogle Scholar
- Ablashi DV, Berneman ZN, Kramarsky B, Whitman J, Asano Y, Pearson GR. Human herpesvirus-7 (HHV-7): current status. Clin Diagn Virol. 1995;4(1):1–13.PubMedView ArticleGoogle Scholar
- Calvanese V, Chavez L, Laurent T, Ding S, Verdin E. Dual-color HIV reporters trace a population of latently infected cells and enable their purification. Virology. 2013;446:283–92.PubMedPubMed CentralView ArticleGoogle Scholar
- Dahabieh MS, Ooms M, Simon V, Sadowski I. A doubly fluorescent HIV-1 reporter shows that the majority of integrated HIV-1 Is latent shortly after infection. J Virol. 2013;87(8):4716–27.PubMedPubMed CentralView ArticleGoogle Scholar
- Sherman E, Nobles C, Berry CC, Six E, Wu Y, Dryga A, et al. INSPIIRED: a pipeline for quantitative analysis of sites of new DNA integration in cellular genomes. Mol Ther Methods Clin Dev. 2017;4:39–49.PubMedView ArticleGoogle Scholar
- Lewin SR, Murray JM, Solomon A, Wightman F, Cameron PU, Purcell DJ, et al. Virologic determinants of success after structured treatment interruptions of antiretrovirals in acute HIV-1 infection. JAIDS J Acquir Immune Defic Syndr. 2008;47(2):140–7.PubMedGoogle Scholar
- Zhang L, Lewin SR, Markowitz M, Lin HH, Skulsky E, Karanicolas R. Measuring recent thymic emigrants in blood of normal and HIV-1-infected individuals before and after effective therapy. J Exp Med. 1999. https://doi.org/10.1084/jem.190.5.725.PubMedPubMed CentralView ArticleGoogle Scholar