scholarly journals The Dual-Specificity Kinase DYRK1A Modulates the Levels of Cyclin L2 To Control HIV Replication in Macrophages

2019 ◽  
Vol 94 (6) ◽  
Author(s):  
Javan K. Kisaka ◽  
Lee Ratner ◽  
George B. Kyei
Keyword(s):  
Public Health Problem ◽  
Critical Factor ◽  
Major Public Health Problem ◽  
Hiv Replication ◽  
Viral Reservoirs ◽  
Dual Specificity ◽  
Cellular Factors ◽  
Latently Infected ◽  
Hiv Eradication ◽  
Hiv 1

ABSTRACT HIV replication in macrophages contributes to the latent viral reservoirs, which are considered the main barrier to HIV eradication. Few cellular factors that facilitate HIV replication in latently infected cells are known. We previously identified cyclin L2 as a critical factor required by HIV-1 and found that depletion of cyclin L2 attenuates HIV-1 replication in macrophages. Here we demonstrate that cyclin L2 promotes HIV-1 replication through interactions with the dual-specificity tyrosine phosphorylation-regulated kinase 1A (DYRK1A). Cyclin L2 and DYRK1A were colocalized in the nucleus and were found together in immunoprecipitation experiments. Knockdown or inhibition of DYRK1A increased HIV-1 replication in macrophages, while depletion of cyclin L2 decreased HIV-1 replication. Furthermore, depletion of DYRK1A increased expression levels of cyclin L2. DYRK1A is a proline-directed kinase that phosphorylates cyclin L2 at serine residues. Mutations of cyclin L2 at serine residues preceding proline significantly stabilized cyclin L2 and increased HIV-1 replication in macrophages. Thus, we propose that DYRK1A controls cyclin L2 expression, leading to restriction of HIV replication in macrophages. IMPORTANCE HIV continues to be a major public health problem worldwide, with over 36 million people living with the virus. Although antiretroviral therapy (ART) can control the virus, it does not provide cure. The virus hides in the genomes of long-lived cells, such as resting CD4+ T cells and differentiated macrophages. To get a cure for HIV, it is important to identify and characterize the cellular factors that control HIV multiplication in these reservoir cells. Previous work showed that cyclin L2 is required for HIV replication in macrophages. However, how cyclin L2 is regulated in macrophages is unknown. Here we show that the protein DYRK1A interacts with and phosphorylates cyclin L2. Phosphorylation makes cyclin L2 amenable to cellular degradation, leading to restriction of HIV replication in macrophages.

scholarly journals Naturally Occurring Compounds Elicit HIV-1 Replication in Chronically Infected Promonocytic Cells

2014 ◽  
Vol 2014 ◽  
pp. 1-6 ◽  
Author(s):  
Andrea Alejandra Barquero ◽  
María Eugenia Dávola ◽  
Diego Ariel Riva ◽  
Susana Esther Mersich ◽  
Laura Edith Alché
Keyword(s):  
Viral Genome ◽  
Nordihydroguaiaretic Acid ◽  
Hiv Replication ◽  
Viral Reservoirs ◽  
Naturally Occurring ◽  
Latently Infected ◽  
Infected Cells ◽  
P24 Antigen ◽  
Hiv 1 ◽  
Promonocytic Cells

Since antiretroviral therapy suppresses but does not eradicate HIV-1 infection, methods to purge viral reservoirs are required. Many strategies involve the reactivation of chronically HIV infected cells to induce the expression of integrated viral genome. In this study, five bioactive compounds, the plant derivatives 1-cinnamoyl-3,11-dihydroxymeliacarpin (CDM), nordihydroguaiaretic acid (NDGA), and curcumin (Cur) and the synthetic stigmasterol analogs (22S,23S)-22,23-dihydroxystigmast-4-en-3-one (compound1) and (22S,23S)-3β-bromo-5α,22,23-trihydroxystigmastan-6-one (compound2), were evaluated for their ability to elicit HIV replication in promonocytic (U1) and lymphocytic (H9+) HIV-1 chronically infected cells. The results revealed that natural compounds CDM, NDGA, and Cur were able to increase HIV-1 p24 antigen, determined by ELISA, only in latently infected promonocytic cells. CDM would reactivate HIV from latency by modulating the release of IL-6 and TNF-α, since the amount of both cytokines measured through ELISA significantly increased in U1 treated cells. Besides, NDGA increased ROS production, which might be related to the increase on p24 level observed in NDGA treated U1. These findings suggest that CDM, NDGA, and Cur might be candidates for further studies on latency-reversing therapeutics to eliminate latently HIV-1 reservoirs.

scholarly journals HIV-1 Vpr mediates the depletion of the cellular repressor CTIP2 to counteract viral gene silencing

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
F. Forouzanfar ◽  
S. Ali ◽  
C. Wallet ◽  
M. De Rovere ◽  
C. Ducloy ◽  
...  
Keyword(s):  
Gene Silencing ◽  
Viral Gene ◽  
Target Cells ◽  
Viral Reservoirs ◽  
Ubiquitin Ligase Complex ◽  
Latently Infected ◽  
Cellular Cofactor ◽  
Proteasome Pathway ◽  
Latent Hiv ◽  
Hiv 1

Abstract Mammals have evolved many antiviral factors impacting different steps of the viral life cycle. Associated with chromatin-modifying enzymes, the cellular cofactor CTIP2 contributes to HIV-1 gene silencing in latently infected reservoirs that constitute the major block toward an HIV cure. We report, for the first time, that the virus has developed a strategy to overcome this major transcriptional block. Productive HIV-1 infection results in a Vpr-mediated depletion of CTIP2 in microglial cells and CD4+ T cells, two of the major viral reservoirs. Associated to the Cul4A-DDB1-DCAF1 ubiquitin ligase complex, Vpr promotes CTIP2 degradation via the proteasome pathway in the nuclei of target cells and notably at the latent HIV-1 promoter. Importantly, Vpr targets CTIP2 associated with heterochromatin-promoting enzymes dedicated to HIV-1 gene silencing. Thereby, Vpr reactivates HIV-1 expression in a microglial model of HIV-1 latency. Altogether our results suggest that HIV-1 Vpr mediates the depletion of the cellular repressor CTIP2 to counteract viral gene silencing.

scholarly journals Purification and Characterization of Oligomeric Envelope Glycoprotein from a Primary R5 Subtype B Human Immunodeficiency Virus

2002 ◽  
Vol 76 (6) ◽  
pp. 2835-2847 ◽  
Author(s):  
Indresh K. Srivastava ◽  
Leonidas Stamatatos ◽  
Harold Legg ◽  
Elaine Kan ◽  
Anne Fong ◽  
...  
Keyword(s):  
Human Immunodeficiency Virus ◽  
Envelope Protein ◽  
Structural Integrity ◽  
Public Health Problem ◽  
Major Public Health Problem ◽  
High Avidity ◽  
Subtype B ◽  
Immunodeficiency Virus ◽  
Hiv 1

ABSTRACT Human immunodeficiency virus (HIV) continues to be a major public health problem throughout the world, with high levels of mortality and morbidity associated with AIDS. Considerable efforts to develop an effective vaccine for HIV have been directed towards the generation of cellular, humoral, and mucosal immune responses. A major emphasis of our work has been toward the evaluation of oligomeric (o-gp140) forms of the HIV type 1 (HIV-1) envelope protein for their ability to induce neutralizing antibody responses. We have derived stable CHO cell lines expressing o-gp140 envelope protein from the primary non-syncytium-inducing (R5) subtype B strain HIV-1US4. We have developed an efficient purification strategy to purify oligomers to near homogeneity. Using a combination of three detectors measuring intrinsic viscosity, light scattering, and refractive index, we calculated the molecular mass of the oligomer to be 474 kDa, consistent with either a trimer or a tetramer. The hydrodynamic radius (Rh ) of o-gp140 was determined to be 8.40 nm, compared with 5.07 nm for the monomer. The relatively smaller Rh of the oligomer suggests that there are indeed differences between the foldings of o-gp140 and gp120. To assess the structural integrity of the purified trimers, we performed a detailed characterization of the glycosylation profile of o-gp140, its ability to bind soluble CD4, and also its ability to bind to a panel of monoclonal antibodies with known epitope specificities for the CD4 binding site, the CD4 inducible site, the V3 loop, and gp41. Immunogenicity studies with rabbits indicated that the purified o-gp140 protein was highly immunogenic and induced high-titer, high-avidity antibodies directed predominantly against conformational epitopes. These observations confirm the structural integrity of purified o-gp140 and its potential as a vaccine antigen.

scholarly journals In vitro anti-HIV activity of ethanol extract from gandarusa (Justicia gendarussa Burm. f) leaves

2020 ◽  
Author(s):  
Ni Putu Ermi Hikmawanti ◽  
Prihartini Widiyanti ◽  
Bambang Prajogo EW
Keyword(s):  
Ethanol Extract ◽  
Hiv Replication ◽  
Acute Hiv Infection ◽  
Hiv Eradication ◽  
Acute Hiv ◽  
P24 Antigen ◽  
Syncytia Formation ◽  
Anti Hiv ◽  
Hiv 1

Anti retroviral drugs for HIV has problems with uncomfortable side effects and that endanger the lives of HIV sufferers. Several herbs have been empirically proven to have an effect on HIV eradication through inhibition of reverse transcriptase. One of such antiviral herbs is Justicia gendarussa (J. gendarussa). The aim of research is to evaluate anti-HIV activity of 70% fractionated-ethanol extract (with releasing alkaloids) and 70% ethanol extract (without releasing alkaloids) of J. gendarussa leaves on in vitro HIV-infected of MOLT-4 cells. The effect of the extracts in inhibiting viral replication and fusion process on acute HIV infection was identi- fied through syncytia formation assay. Effect of the extracts on HIV p24 antigen was evaluated using HIV-1 p24 ELISA kit. It was found that 70% fractionated-ethanol extract and 70% ethanol extract of J. gendarussa leaves significantly inhibited of HIV replication by inhibition of syncytia formation, where the 50% effective concen- tration (EC50) values of the 70% fractionated-ethanol extract and 70% ethanol extract are 70.5 μg/mL and 228.7 μg/mL, respec- tively. Both of the extracts were also significantly inhibited HIV replication by decreasing HIV p24 antigen level where the EC 50 values of the 70% fractionated-ethanol extract and 70% ethanol extract are 88.8 μg/mL and 540.7 μg/mL, respectively. Moreover, it was found that 70% fractionated-ethanol extract of J. gendarussa leaves has anti-HIV activity since its EC50 values less than 100 μg/mL. It was concluded that J. gendarussa could be potentially developed into a phytopharmaceutical product due to its anti-HIV activity.

scholarly journals FISHing Out the Hidden Enemy: Advances in Detecting and Measuring Latent HIV-Infected Cells

mBio ◽  
2017 ◽  
Vol 8 (5) ◽  
Author(s):  
Vinayaka R. Prasad ◽  
Ganjam V. Kalpana
Keyword(s):  
Immune Surveillance ◽  
Gag Protein ◽  
Link Type ◽  
Latently Infected ◽  
Hiv Eradication ◽  
Infected Cells ◽  
Latent Hiv ◽  
Cell Host Microbe ◽  
Unique Cell ◽  
Hiv 1

ABSTRACT The indomitable aspect of HIV-1 infection is not that HIV-1 proviral DNA is integrated into host DNA but that it can also turn itself off, remaining invisible to drug or immune surveillance. Thus, the goals of eradication include ways to precisely excise HIV-1 DNA or wake up the silent HIV-1 provirus and eliminate the infected cells thus identified. Methods to identify and fish out the latently infected cells or to delineate their characteristics are being rapidly developed. In 2016, Baxter et al. (A. E. Baxter, J. Niessl, R. Fromentin, J. Richard, F. Porichis, R. Charlebois, M. Massanella, N. Brassard, N. Alsahafi, G. G. Delgado, J. P. Routy, B. D. Walker, A. Finzi, N. Chomont, and D. E. Kaufmann, Cell Host Microbe 20:368–380, 2016, https://doi.org/10.1016/j.chom.2016.07.015 ) and Martrus et al. (G. Martrus, A. Niehrs, R. Cornelis, A. Rechtien, W. García-Beltran, M. Lütgehetmann, C. Hoffmann, and M. Altfeld, J Virol 90:9018–9028, 2016, https://doi.org/10.1128/JVI.01448-16 ) reported using the fluorescence in situ hybridization-flow cytometry technique to identify and quantify cells expressing HIV-1 RNA and Gag protein, as well as bearing unique cell surface markers. In a recent article in mBio, Grau-Expósito et al. (J. Grau-Expósito, C. Serra-Peinado, L. Miguel, J. Navarro, A. Curran, J. Burgos, I. Ocaña, E. Ribera, A. Torrella, B. Planas, R. Badía, J. Castellví, V. Falcó, M. Crespo, and M. J. Buzon, mBio 8:e00876-17, 2017, https://doi.org/10.1128/mBio.00876-17 !) reported a similar method that they claim to be more sensitive. With these methods, researchers are one step closer to measuring latent reservoirs and eliminating critical barriers to HIV eradication.

scholarly journals Higher Native Peruvian ancestry proportion is associated with tuberculosis progression risk

2020 ◽  
Author(s):  
Samira Asgari ◽  
Yang Luo ◽  
Kamil Slowikowski ◽  
Chuan-Chin Huang ◽  
Roger Calderon ◽  
...  
Keyword(s):  
Risk Factors ◽  
Genetic Risk ◽  
Public Health Problem ◽  
Genetic Risk Factors ◽  
Indigenous Populations ◽  
Major Public Health Problem ◽  
Road Map ◽  
Increased Risk ◽  
Latently Infected ◽  
Progression Risk

The global burden of pulmonary tuberculosis (TB) remains a major public health problem that is particularly severe among and disproportionately affects indigenous populations. We aimed to investigate whether genetic factors related to indigeneity affect TB progression risk in a cohort of admixed Peruvians with active TB and their latently infected household contacts. Our results show that Native Peruvian ancestry is positively associated with TB progression risk: a 10% increase in native ancestry tracks with a 25% increased risk of TB progression. This risk is independent of the potentially confounding socio-demographic and environmental factors that we tested here. Our results demonstrate that the genetic contribution to TB risk varies among populations and brings new insight to the long-standing debate on the role of genetic ancestry in susceptibility to TB. Additionally, our study highlights the value of including diverse populations in genetic studies of infectious diseases and other complex phenotypes, and provides a road map for future similar studies where it is important to account for confounding non-genetic risk factors to identify genetic risk factors.

Novel Phorbol Esters Exert Dichotomous Effects on Inhibition of HIV-1 Infection and Activation of Latent HIV-1 Expression

2005 ◽  
Vol 16 (5) ◽  
pp. 303-313
Author(s):  
Yu Zhong ◽  
Yuji Matsuya ◽  
Hideo Nemoto ◽  
Masao Mori ◽  
Haruo Saito ◽  
...  
Keyword(s):  
Mtt Assay ◽  
Mononuclear Cells ◽  
Phorbol Esters ◽  
High Concentration ◽  
Viral Reservoirs ◽  
Peripheral Blood Mononuclear ◽  
Latently Infected ◽  
Low Cytotoxicity ◽  
Latent Hiv ◽  
Hiv 1

Two new phorbol esters, NPB-11 (12- O-methoxymethylphorbol-13-decanoate) and NPB-15 (12- O-benzyloxymethylphorbol-13-decanoate) were synthesized. The compounds exhibited potent anti-HIV-1 activity and low cytotoxicity in MT-4 cells by MTT assay even at a high concentration [50% cytotoxic concentrations (CC50) were 8.32 and 4.39 μg/ml, respectively]. Two inhibitors strongly suppressed HIV-1 (IIIB strain) replication in MT-4 cells with a 50% effective concentration (EC50) of 1.3 and 0.27 ng/ml, respectively. NPB-11 efficiently blocked replication of both X4 and R5 HIV-1 in PHA-activated peripheral blood mononuclear cells and MT-4 cells as revealed by p24 assay. The antiviral activity appeared to be mediated, at least partially, by the down-regulation of the expression of CD4 and the HIV-1 co-receptors, CXCR4 and CCR5. The compounds were also capable of selectively up-regulating HIV-1 expression in a variety of latently infected cell lines and inducing cell death in HIV-1 infected cells. The effect of NPBs on the induction of HIV-1 was specifically blocked by nontoxic doses of a protein kinase C blocker, staurosporine. NPB-11 blocked the spread of HIV-1 released from latently infected ACH-2 cells to MT-4 cells in a co-culture system. When combined with AZT, NPB-11 synergistically inhibited HIV-1 replication in MTT assay using MT-4 cells. These data suggest that these agents might be useful in reducing persistent viral reservoirs in patients and as adjuvant therapy in patients treated with HAART.

scholarly journals Low-Level Ionizing Radiation Induces Selective Killing of HIV-1-Infected Cells with Reversal of Cytokine Induction Using mTOR Inhibitors

Viruses ◽  
2020 ◽  
Vol 12 (8) ◽  
pp. 885
Author(s):  
Daniel O. Pinto ◽  
Catherine DeMarino ◽  
Thy T. Vo ◽  
Maria Cowen ◽  
Yuriy Kim ◽  
...  
Keyword(s):  
Ionizing Radiation ◽  
Mtor Inhibitors ◽  
Viral Reservoirs ◽  
Viral Loads ◽  
Viral Spread ◽  
Novel Approach ◽  
Latently Infected ◽  
Shock And Kill ◽  
Infected Cells ◽  
Hiv 1

HIV-1 infects 39.5 million people worldwide, and cART is effective in preventing viral spread by reducing HIV-1 plasma viral loads to undetectable levels. However, viral reservoirs persist by mechanisms, including the inhibition of autophagy by HIV-1 proteins (i.e., Nef and Tat). HIV-1 reservoirs can be targeted by the “shock and kill” strategy, which utilizes latency-reversing agents (LRAs) to activate latent proviruses and immunotarget the virus-producing cells. Yet, limitations include reduced LRA permeability across anatomical barriers and immune hyper-activation. Ionizing radiation (IR) induces effective viral activation across anatomical barriers. Like other LRAs, IR may cause inflammation and modulate the secretion of extracellular vesicles (EVs). We and others have shown that cells may secrete cytokines and viral proteins in EVs and, therefore, LRAs may contribute to inflammatory EVs. In the present study, we mitigated the effects of IR-induced inflammatory EVs (i.e., TNF-α), through the use of mTOR inhibitors (mTORi; Rapamycin and INK128). Further, mTORi were found to enhance the selective killing of HIV-1-infected myeloid and T-cell reservoirs at the exclusion of uninfected cells, potentially via inhibition of viral transcription/translation and induction of autophagy. Collectively, the proposed regimen using cART, IR, and mTORi presents a novel approach allowing for the targeting of viral reservoirs, prevention of immune hyper-activation, and selectively killing latently infected HIV-1 cells.

scholarly journals Th17 cell master transcription factor RORC2 regulates HIV-1 gene expression and viral outgrowth

2021 ◽  
Vol 118 (48) ◽  
pp. e2105927118
Author(s):  
Tomas Raul Wiche Salinas ◽  
Yuwei Zhang ◽  
Daniele Sarnello ◽  
Alexander Zhyvoloup ◽  
Laurence Raymond Marchand ◽  
...  
Keyword(s):  
Gene Expression ◽  
T Cells ◽  
Th17 Cell ◽  
Th17 Cells ◽  
Nuclear Hormone Receptor ◽  
Long Terminal Repeats ◽  
T Helper 17 ◽  
Hiv Replication ◽  
Latently Infected ◽  
Hiv 1

Among CD4+ T cells, T helper 17 (Th17) cells are particularly susceptible to HIV-1 infection and are depleted from mucosal sites, which causes damage to the gut barrier, resulting in a microbial translocation-induced systemic inflammation, a hallmark of disease progression. Furthermore, a proportion of latently infected Th17 cells persist long term in the gastrointestinal lymphatic tract where a low-level HIV-1 transcription is observed. This residual viremia contributes to chronic immune activation. Thus, Th17 cells are key players in HIV pathogenesis and viral persistence. It is, however, unclear why these cells are highly susceptible to HIV-1 infection. Th17 cell differentiation depends on the expression of the master transcriptional regulator RORC2, a retinoic acid-related nuclear hormone receptor that regulates specific transcriptional programs by binding to promoter/enhancer DNA. Here, we report that RORC2 is a key host cofactor for HIV replication in Th17 cells. We found that specific inhibitors that bind to the RORC2 ligand-binding domain reduced HIV replication in CD4+ T cells. The depletion of RORC2 inhibited HIV-1 infection, whereas its overexpression enhanced it. RORC2 was also found to promote HIV-1 gene expression by binding to the nuclear receptor responsive element in the HIV-1 long terminal repeats (LTR). In treated HIV-1 patients, RORC2+ CD4 T cells contained more proviral DNA than RORC2− cells. Pharmacological inhibition of RORC2 potently reduced HIV-1 outgrowth in CD4+ T cells from antiretroviral-treated patients. Altogether, these results provide an explanation as to why Th17 cells are highly susceptible to HIV-1 infection and suggest that RORC2 may be a cell-specific target for HIV-1 therapy.

scholarly journals Immune profiling in M. tuberculosis infection enables stratification of patients with active disease

2019 ◽  
Author(s):  
Darragh Duffy ◽  
Elisa Nemes ◽  
Alba Llibre ◽  
Vincent Rouilly ◽  
Elizabeth Filander ◽  
...  
Keyword(s):  
Whole Blood ◽  
Public Health Problem ◽  
Blood Collection ◽  
Major Public Health Problem ◽  
Antigen Stimulation ◽  
Early Treatment Response ◽  
Before And After ◽  
Latently Infected ◽  
Response Current ◽  
Active Disease

AbstractTuberculosis (TB) is caused byMycobacterium tuberculosis(Mtb) infection and is a major public health problem with an estimated 1.7 billion persons infected worldwide. Clinical challenges in TB include the lack of a blood-based test for active disease, and the absence of prognostic biomarkers for early treatment response. Current blood based tests, such as QuantiFERON-TB Gold (QFT), are based on an IFNγ readout followingMtbantigen stimulation. However, they do not distinguish active TB disease from asymptomaticMtbinfection. We hypothesized that the use of TruCulture, an improved immunomonitoring method for whole blood collection and immune stimulation, could improve the discrimination of active disease from latentMtbinfection. To test our hypothesis, we stimulated whole blood from active TB patients (before and after successful treatment), comparing them to asymptomatic latently infected individuals.Mtb-specific antigens (ESAT-6, CFP-10, TB7.7) and live bacillus Calmette-Guerin (BCG) were used for TruCulture stimulation conditions, with direct comparison to QFT. Protein analyses were performed on the culture supernatants using ELISA and Luminex multi-analyte profiling. TruCulture showed an ability to discriminate active TB cases from latent controls (p < 0.0001, AUC = 0.81, 95% CI: 0.69-0.93) as compared to QFT (p = 0.47 AUC = 0.56, 95% CI: 0.40-0.72), based on an IFNγ readout afterMtbantigen stimulation. The stratification of the two groups could be further improved by using theMtbAg/BCG IFNγ ratio response (p < 0.0001, AUC = 0.918, 95% CI: 0.84-0.98). We also identified additional cytokines that distinguished latent infection from TB disease; and show that the primary differences between the TruCulture and QFT systems were a result of higher levels of non-specific innate immune activation in QFT tubes, due to the lack of a buffering solution in the latter. We conclude that TruCulture offers a next-generation solution for whole blood stimulation and immunomonitoring with the possibility to discriminate active and latently infected persons.

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