Potent and Selective Inhibition of Tat-Dependent HIV-1 Replication in Chronically Infected Cells by a Novel Naphthalene Derivative JTK-101

In search for effective human immunodeficiency virus type 1 (HIV-1) transcription inhibitors, we have evaluated more than 100,000 compounds for their inhibitory effects on HIV-1 long terminal repeat (LTR)-driven reporter gene expression, and identified a novel naphthalene derivative, JTK-101. This compound could suppress tumour necrosis factor (TNF)-α-induced HIV-1 production in latently infected OM-10.1 cells at nanomolar concentrations. JTK-101 could also potently inhibit constitutive HIV-1 production in MOTL-4/IIIB. However, the antiviral activity of JTK-101 was found to be much weaker in acutely infected cells and the chronically infected cells U937/IIIB cells than in OM-10.1 and MOLT-4/IIIB cells. JTK-101 selectively suppressed TNF-α-induced HIV-1 mRNA synthesis in OM-10.1 cells in a dose-dependent fashion. JTK-101 modestly inhibited TNF-α-induced HIV-1 LTR-driven reporter gene expression, but potently inhibited Tat-induced gene expression. Immunoblot analysis revealed that low-level expression of the Tat cofactors CDK9 and cyclin T1 might contribute to the diminished antiviral activity in U937/IIIB cells. Furthermore, JTK-101 could not inhibit HIV-1 replication in chronically infected monocytes/macrophages, in which CDK9 and cyclin T1 were undetectable. These results suggest that JTK-101 exerts its anti-HIV-1 activity through the inhibition of known or unknown Tat cofactors, presumably CDK9/cyclin T1.

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Inhibition of Human Immunodeficiency Virus Type 1 Replication in Acutely and Chronically Infected Cells by EM2487, a Novel Substance Produced by a Streptomyces Species

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.43.10.2350 ◽

1999 ◽

Vol 43 (10) ◽

pp. 2350-2355 ◽

Cited By ~ 16

Author(s):

Masanori Baba ◽

Mika Okamoto ◽

Hitoshi Takeuchi

Keyword(s):

Gene Expression ◽

Human Immunodeficiency Virus ◽

Reporter Gene ◽

Human Immunodeficiency Virus Type ◽

Virus Type ◽

Tnf Α ◽

Immunodeficiency Virus ◽

Infected Cells ◽

Hiv 1

ABSTRACT In a search for effective HIV-1 transcription inhibitors, we have evaluated more than 75,000 compounds for their inhibitory effects on Tat-induced human immunodeficiency virus type 1 (HIV-1) long terminal repeat (LTR)-driven reporter gene expression and found that EM2487, a novel small-molecule substance produced by a Streptomycesspecies, is a potent and selective inhibitor of HIV-1 replication in both acutely and chronically infected cells. Its 50% effective concentration for acute HIV-1 infection was 0.27 μM in peripheral blood mononuclear cells (PBMCs), while the 50% cytotoxic concentration for mock-infected PBMCs was 13.3 μM. EM2487 proved inhibitory to a variety of HIV-1 strains and HIV-2 in acutely infected T-cell lines (MOLT-4 and MT-4). The compound could suppress tumor necrosis factor alpha (TNF-α)-induced HIV-1 production in latently infected cells (OM-10.1 and ACH-2) as well as constitutive viral production in chronically infected cells (MOLT-4/IIIB and U937/IIIB) without showing any cytotoxicity. EM2487 did not affect early events of the HIV-1 replication cycle, as determined by proviral DNA synthesis in acutely infected MOLT-4 cells. In contrast, the compound selectively prevented viral mRNA synthesis in OM-10.1 cells, suggesting that HIV-1 inhibition occurs at the transcriptional level. Furthermore, EM2487 did not inhibit TNF-α-induced HIV-1 LTR-driven reporter gene expression but did inhibit that induced by Tat, irrespective of the presence or absence of the nuclear factor κB binding sites in the LTR. These results suggest that the mechanism of action is attributable in part to the inhibition of Tat function.

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273. Reporter Gene Expression Following Repeat Administration of a HIV-1 Lentiviral Vector in Mouse Airways

Molecular Therapy ◽

10.1016/s1525-0016(16)36845-9 ◽

2011 ◽

Vol 19 ◽

pp. S106

Keyword(s):

Gene Expression ◽

Reporter Gene ◽

Lentiviral Vector ◽

Reporter Gene Expression ◽

Repeat Administration ◽

Hiv 1

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CG dinucleotide removal in bioluminescent and fluorescent reporters improves HIV-1 replication and reporter gene expression for dual imaging in humanized mice

Journal of Virology ◽

10.1128/jvi.00449-21 ◽

2021 ◽

Author(s):

Chandra N. Roy ◽

Mariana A. Benitez Moreno ◽

Chris Kline ◽

Zandrea Ambrose

Keyword(s):

Gene Expression ◽

Reporter Gene ◽

Reporter Genes ◽

Reporter Gene Expression ◽

Humanized Mice ◽

Plasma Viremia ◽

Reporter Expression ◽

Cg Dinucleotides ◽

Hiv 1

Visualizing the transmission and dissemination of human immunodeficiency virus type 1 (HIV-1) in real time in humanized mouse models is a robust tool to investigate viral replication during treatments and in tissue reservoirs. However, the stability and expression of HIV-1 reporter genes are obstacles for long-term serial imaging in vivo . Two replication-competent CCR5-tropic HIV-1 reporter constructs were created that encode either nanoluciferase (nLuc) or a near - infrared fluorescent protein (iRFP) upstream of nef . HIV-1 reporter virus replication and reporter gene expression was measured in cell culture and in humanized mice. While reporter gene expression in vivo correlated initially with plasma viremia, expression decreased after 4-5 weeks despite high plasma viremia. The reporter genes were codon-optimized to remove cytosine/guanine (CG) dinucleotides and new CO-nLuc and CO-iRFP viruses were reconstructed. Removal of CG dinucleotides in HIV-1 reporter viruses improved replication in vitro and reporter expression in vivo and ex vivo . Both codon optimized reporter viruses could be visualized during co-infection and in vivo reporter gene expression during treatment failure preceded detection of plasma viremia. While the dynamic range of CO-iRFP HIV-1 was lower than that of CO-nLuc HIV-1, both viruses could have utility in studying and visualizing HIV-1 infection in humanized mice. Importance Animal models are important for studying HIV-1 pathogenesis and treatments. We developed two viruses each encoding a reporter gene that can be expressed in cells after infection. This study shows that HIV-1 infection can be visualized by noninvasive, whole body imaging in mice with human immune cells over time by reporter expression. We improved reporter expression to reflect HIV-1 replication and showed that two viral variants can be tracked over time in the same animal and can predict failure of antiretroviral therapy to suppress virus.

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