HIV-1 can infect northern pig-tailed macaques ( Macaca leonina ) and form viral reservoirs in vivo

ABSTRACT The molecular basis for HIV-1 susceptibility in primary human monocyte-derived macrophages (MDMs) was previously evaluated by comparing the transcriptome of infected and bystander populations. Careful analysis of the data suggested that the ubiquitin ligase MDM2 acted as a positive regulator of HIV-1 replication in MDMs. In this study, MDM2 silencing through transcript-specific small interfering RNAs in MDMs induced a reduction in HIV-1 reverse transcription and integration along with an increase in the expression of p53-induced genes, including CDKN1A. Experiments with Nutlin-3, a pharmacological inhibitor of MDM2 p53-binding activity, showed a similar effect on HIV-1 infection, suggesting that the observed restriction in HIV-1 production results from the release/activation of p53 and not the absence of MDM2 per se. Knockdown and inhibition of MDM2 also both correlate with a decrease in the Thr592-phosphorylated inactive form of SAMHD1. The expression level of MDM2 and the p53 activation status are therefore important factors in the overall susceptibility of macrophages to HIV-1 infection, bringing a new understanding of signaling events controlling the process of virus replication in this cell type. IMPORTANCE Macrophages, with their long life span in vivo and their resistance to HIV-1-mediated cytopathic effect, might serve as viral reservoirs, contributing to virus persistence in an infected individual. Identification of host factors that increase the overall susceptibility of macrophages to HIV-1 might provide new therapeutic targets for the efficient control of viral replication in these cells and limit the formation of reservoirs in exposed individuals. In this study, we demonstrate the importance of p53 regulation by MDM2, which creates a cellular environment more favorable to the early steps of HIV-1 replication. Moreover, we show that p53 stabilization reduces virus infection in human macrophages, highlighting the important role of p53 in antiviral immunity.

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An ultrasensitive planar array p24 Gag ELISA to detect individual HIV-1 viral particles and infected cells

10.21203/rs.3.rs-832339/v1 ◽

2021 ◽

Author(s):

Alberto Bosque ◽

Callie Levinger ◽

J Natalie Howard ◽

Pingtao Tang ◽

Amit Joshi

Keyword(s):

Biological Fluids ◽

Low Frequency ◽

Viral Reservoirs ◽

Hiv Persistence ◽

Viral Particles ◽

Planar Array ◽

Infected Cells ◽

Major Loss ◽

Hiv 1

Abstract Human Immunodeficiency virus-1 (HIV-1) persistence in the presence of antiretroviral therapy (ART) has halted the development of curative strategies. Measuring HIV persistence is complex due to the low frequency of cells containing virus in vivo. Most of the commercially available assays to date measure nucleic acid. These assays have the advantage of being highly sensitive and allow for the analysis of sequence diversity, intactness of the HIV genome or evaluation of diverse RNA species. However, these assays are limited in evaluating translational competent viral reservoirs. In here, we developed an ultrasensitive p24 ELISA that uses the SimoaTM planar array technology that can detect as low as a single HIV-1 particle and a single HIV-1 infected cell. Furthermore, the assay is optimized to measure very low levels of p24 in different biological fluids without a major loss of sensitivity or reproducibility. Our results demonstrate that the ‘homebrew' planar p24 ELISA immunoassay is a broadly applicable new tool to evaluate HIV persistence in diverse biological fluids.

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HIV-1 and microglia: EcoHIV and HIV-1 transgenic rats

10.1101/2020.11.03.365494 ◽

2020 ◽

Author(s):

Hailong Li ◽

Kristen A. McLaurin ◽

Jessica M. Illenberger ◽

Charles F. Mactutus ◽

Rosemarie M. Booze

Keyword(s):

Distribution Pattern ◽

Biological System ◽

Cell Type ◽

Viral Reservoirs ◽

Major Cell Type ◽

Key Aspects ◽

The Brain ◽

Hiv 1

ABSTRACTThe persistence of HIV-1 viral reservoirs in the brain, despite treatment with combination antiretroviral therapy (cART), remains a critical roadblock for the development of a novel cure strategy for HIV-1. To enhance our understanding of viral reservoirs, two complementary studies were conducted to 1) evaluate the HIV-1 mRNA neuroanatomical distribution pattern and major cell type expressing HIV-1 mRNA in the HIV-1 transgenic (Tg) rat (i.e., under conditions of latent infection), and 2) to validate our findings by developing and critically testing a novel biological system to model active HIV-1 infection in the rat. First, a restricted, region-specific HIV-1 mRNA distribution pattern was observed in the HIV-1 Tg rat. Microglia were the predominant cell type expressing HIV-1 mRNA in the HIV-1 Tg rat. Second, we developed and critically tested a novel biological system to model key aspects of HIV-1 by infusing F344/N control rats with chimeric HIV (EcoHIV). In vitro, primary cultured microglia were treated with EcoHIV revealing prominent expression within 24 hours of infection. In vivo, EcoHIV expression was observed seven days after stereotaxic injections. Following EcoHIV infection, microglia were the major cell type expressing HIV-1 mRNA, results which are consistent with observations in the HIV-1 Tg rat. Within eight weeks of infection, EcoHIV rats exhibited neurocognitive impairments, synaptic dysfunction, which may result from activation of the NogoA-NgR3/PirB-RhoA signaling pathway, and neuroinflammation. Collectively, these studies enhance our understanding of HIV-1 viral reservoirs in the brain and offer a novel biological system to model HIV-associated neurocognitive disorders and associated comorbidities (i.e., drug abuse) in rats.

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Microglial HIV-1 Expression: Role in HIV-1 Associated Neurocognitive Disorders

Viruses ◽

10.3390/v13050924 ◽

2021 ◽

Vol 13 (5) ◽

pp. 924

Author(s):

Hailong Li ◽

Kristen A. McLaurin ◽

Jessica M. Illenberger ◽

Charles F. Mactutus ◽

Rosemarie M. Booze

Keyword(s):

Distribution Pattern ◽

Biological System ◽

Neurocognitive Disorders ◽

Cell Type ◽

Viral Reservoirs ◽

Major Cell Type ◽

The Brain ◽

Hiv 1

The persistence of HIV-1 viral reservoirs in the brain, despite treatment with combination antiretroviral therapy (cART), remains a critical roadblock for the development of a novel cure strategy for HIV-1. To enhance our understanding of viral reservoirs, two complementary studies were conducted to (1) evaluate the HIV-1 mRNA distribution pattern and major cell type expressing HIV-1 mRNA in the HIV-1 transgenic (Tg) rat, and (2) validate our findings by developing and critically testing a novel biological system to model active HIV-1 infection in the rat. First, a restricted, region-specific HIV-1 mRNA distribution pattern was observed in the HIV-1 Tg rat. Microglia were the predominant cell type expressing HIV-1 mRNA in the HIV-1 Tg rat. Second, we developed and critically tested a novel biological system to model key aspects of HIV-1 by infusing F344/N control rats with chimeric HIV (EcoHIV). In vitro, primary cultured microglia were treated with EcoHIV revealing prominent expression within 24 h of infection. In vivo, EcoHIV expression was observed seven days after stereotaxic injections. Following EcoHIV infection, microglia were the major cell type expressing HIV-1 mRNA, results that are consistent with observations in the HIV-1 Tg rat. Within eight weeks of infection, EcoHIV rats exhibited neurocognitive impairments and synaptic dysfunction, which may result from activation of the NogoA-NgR3/PirB-RhoA signaling pathway and/or neuroinflammation. Collectively, these studies enhance our understanding of HIV-1 viral reservoirs in the brain and offer a novel biological system to model HIV-associated neurocognitive disorders and associated comorbidities (i.e., drug abuse) in rats.

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An ultrasensitive planar array p24 Gag ELISA to detect HIV-1 in diverse biological matrixes

Scientific Reports ◽

10.1038/s41598-021-03072-7 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Callie Levinger ◽

JNatalie Howard ◽

Jie Cheng ◽

Pingtao Tang ◽

Amit Joshi ◽

...

Keyword(s):

Nucleic Acid ◽

Limit Of Detection ◽

Biological Fluids ◽

Low Frequency ◽

Viral Reservoirs ◽

Hiv Persistence ◽

Planar Array ◽

Major Loss ◽

Hiv 1

AbstractHuman immunodeficiency virus-1 (HIV-1) persistence in the presence of antiretroviral therapy (ART) has halted the development of curative strategies. Measuring HIV persistence is complex due to the low frequency of cells containing virus in vivo. Most of the commercially available assays to date measure nucleic acid. These assays have the advantage of being highly sensitive and allow for the analysis of sequence diversity, intactness of the HIV genome or evaluation of diverse RNA species. However, these assays are limited in evaluating translational competent viral reservoirs. In here, we developed an ultrasensitive p24 ELISA that uses the Simoa planar array technology that can detect HIV-1 virions and HIV-1 infected cell with limit of detection similar to nucleic acid assays. Furthermore, the assay is optimized to measure very low levels of p24 in different biological fluids without a major loss of sensitivity or reproducibility. Our results demonstrate that the ‘homebrew’ planar p24 ELISA immunoassay is a broadly applicable new tool to evaluate HIV persistence in diverse biological fluids and cells.

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Multi-organ Targeting of HIV-1 Viral Reservoirs with Etravirine Loaded Nanostructured Lipid Carrier: An In-vivo Proof of Concept

European Journal of Pharmaceutical Sciences ◽

10.1016/j.ejps.2021.105916 ◽

2021 ◽

pp. 105916

Author(s):

Satish Rojekar ◽

Leila Fotooh ◽

Rohan Pai ◽

Ketan Mahajan ◽

Smita Kulkarni ◽

...

Keyword(s):

Nanostructured Lipid Carrier ◽

Proof Of Concept ◽

Viral Reservoirs ◽

Organ Targeting ◽

Hiv 1

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Antiretroviral Therapy in Macrophages: Implication for HIV Eradication

Antiviral Chemistry and Chemotherapy ◽

10.3851/imp1374 ◽

2009 ◽

Vol 20 (2) ◽

pp. 63-78 ◽

Cited By ~ 62

Author(s):

Christina Gavegnano ◽

Raymond F Schinazi

Keyword(s):

Antiretroviral Therapy ◽

Antiviral Activity ◽

Productive Infection ◽

Viral Reservoir ◽

Viral Reservoirs ◽

Hiv Therapy ◽

Cellular Pharmacology ◽

Latently Infected ◽

Hiv 1

HIV type-1 (HIV-1) accounts for more than 25 million deaths and nearly 40 million people are infected worldwide. A significant obstacle in clearing virus from infected individuals is latently infected viral reservoirs. Latent HIV-1 can emerge with recrudescence as a productive infection later in disease progression and could provide a source for the emergence of resistant HIV-1. It is widely recognized that macrophages represent a latently infected viral reservoir and are a significant and critical HIV-1 target cell in vivo. Macrophages can be divided into multiple subsets of macrophage-like cells, all of which are susceptible to HIV-1 infection, including dendritic cells, Langerhans cells, alveolar macrophages, mucosal macrophages and microglial cells. Current antiretroviral therapy (ART) often displays differential antiviral activity in macrophages relative to CD4+ T-lymphocytes. Significant work has been performed to establish antiviral activity of many clinically approved ART in macrophages; however, a direct link between antiviral activity and specific mechanisms responsible for these antiviral effects are incompletely understood. This review identifies many understudied areas of research, along with topics for further research in the field of HIV therapy and eradication. Discussion focuses upon the known cellular pharmacology and antiviral activity of antiretroviral agents in macrophages and its relationship to latency, chronic HIV-1 infection and therapeutic strategies to eradicate systemic HIV-1 infection.

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Potential Utilization of APOBEC3-Mediated Mutagenesis for an HIV-1 Functional Cure

Frontiers in Microbiology ◽

10.3389/fmicb.2021.686357 ◽

2021 ◽

Vol 12 ◽

Author(s):

Terumasa Ikeda ◽

Yuan Yue ◽

Ryo Shimizu ◽

Hesham Nasser

Keyword(s):

T Cells ◽

Cytotoxic T Lymphocyte ◽

Mutagenic Activity ◽

Primary Source ◽

Virus Infectivity ◽

Functional Cure ◽

Viral Reservoirs ◽

Proviral Dna ◽

Hiv 1

The introduction of combination antiretroviral therapy (cART) has managed to control the replication of human immunodeficiency virus type 1 (HIV-1) in infected patients. However, a complete HIV-1 cure, including a functional cure for or eradication of HIV-1, has yet to be achieved because of the persistence of latent HIV-1 reservoirs in adherent patients. The primary source of these viral reservoirs is integrated proviral DNA in CD4+ T cells and other non-T cells. Although a small fraction of this proviral DNA is replication-competent and contributes to viral rebound after the cessation of cART, >90% of latent viral reservoirs are replication-defective and some contain high rates of G-to-A mutations in proviral DNA. At least in part, these high rates of G-to-A mutations arise from the APOBEC3 (A3) family proteins of cytosine deaminases. A general model has shown that the HIV-1 virus infectivity factor (Vif) degrades A3 family proteins by proteasome-mediated pathways and inactivates their antiviral activities. However, Vif does not fully counteract the HIV-1 restriction activity of A3 family proteins in vivo, as indicated by observations of A3-mediated G-to-A hypermutation in the proviral DNA of HIV-1-infected patients. The frequency of A3-mediated hypermutation potentially contributes to slower HIV-1/AIDS disease progression and virus evolution including the emergence of cytotoxic T lymphocyte escape mutants. Therefore, combined with other strategies, the manipulation of A3-mediated mutagenesis may contribute to an HIV-1 functional cure aimed at cART-free remission. In this mini-review, we discuss the possibility of an HIV-1 functional cure arising from manipulation of A3 mutagenic activity.

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