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Dengue trafficking

Fabio Zanini, Szu-Yuan Pu, Elena Bekerman, Shirit Einav, Stephen R Quake
Dengue and Zika viral infections affect millions of people annually and can be complicated by hemorrhage or neurological manifestations, respectively. However, a thorough understanding of the host response to these viruses is lacking, partly because conventional approaches ignore heterogeneity in virus abundance across cells. We present viscRNA-Seq (virus-inclusive single cell RNA-Seq), an approach to probe the host transcriptome together with intracellular viral RNA at the single cell level. We applied viscRNA-Seq to monitor dengue and Zika virus infection in cultured cells and discovered extreme heterogeneity in virus abundance...
February 16, 2018: ELife
Rassamon Keawvichit, Ladawan Khowawisetsut, Sakaorat Lertjuthaporn, Kanchana Tangnararatchakit, Nopporn Apiwattanakul, Sutee Yoksan, Ampaiwan Chuansumrit, Kulkanya Chokephaibulkit, Aftab A Ansari, Nattawat Onlamoon, Kovit Pattanapanyasat
Dengue virus (DENV) infection is considered as one of the most important mosquito-borne diseases. It causes a spectrum of illness that could be due to qualitative and/or quantitative difference(s) of the natural killer (NK)-cell responses during acute DENV infection. This view prompted us to perform a detailed phenotypic comparative characterization of NK cell subsets from DENV-infected patients with dengue fever (DF), dengue hemorrhagic fever (DHF) and healthy controls. The activation/differentiation molecules, CD69 and CD57 and a variety of tissue homing molecules were analyzed on the CD56(hi) CD16(-) and CD56(lo) CD16(+) NK cells...
November 4, 2017: Immunology
Vincent Geoghegan, Kirsty Stainton, Stephanie M Rainey, Thomas H Ant, Adam A Dowle, Tony Larson, Svenja Hester, Philip D Charles, Benjamin Thomas, Steven P Sinkins
Wolbachia are intracellular maternally inherited bacteria that can spread through insect populations and block virus transmission by mosquitoes, providing an important approach to dengue control. To better understand the mechanisms of virus inhibition, we here perform proteomic quantification of the effects of Wolbachia in Aedes aegypti mosquito cells and midgut. Perturbations are observed in vesicular trafficking, lipid metabolism and in the endoplasmic reticulum that could impact viral entry and replication...
September 13, 2017: Nature Communications
Sineewanlaya Wichit, Rodolphe Hamel, Eric Bernard, Loïc Talignani, Fodé Diop, Pauline Ferraris, Florian Liegeois, Peeraya Ekchariyawat, Natthanej Luplertlop, Pornapat Surasombatpattana, Frédéric Thomas, Andres Merits, Valérie Choumet, Pierre Roques, Hans Yssel, Laurence Briant, Dorothée Missé
Chikungunya virus (CHIKV) is an emerging arbovirus of the Togaviridae family that poses a present worldwide threat to human in the absence of any licensed vaccine or antiviral treatment to control viral infection. Here, we show that compounds interfering with intracellular cholesterol transport have the capacity to inhibit CHIKV replication in human skin fibroblasts, a major viral entry site in the human host. Pretreatment of these cells with the class II cationic amphiphilic compound U18666A, or treatment with the FDA-approved antidepressant drug imipramine resulted in a near total inhibition of viral replication and production at the highest concentration used without any cytotoxic effects...
June 9, 2017: Scientific Reports
Sivan Padma Priya, S Sakinah, Mok Pooi Ling, Hui-Yee Chee, Akon Higuchi, Rukman Awang Hamat, Vasantha Kumari Neela, Abdullah A Alarfaj, Murugan A Munusamy, Ashraf A Hatamleh, Ahmed E Al-Sabri, Ibrahim Ahmad Abdulaziz Al-Suwailem, Mariappan Rajan, Giovanni Benelli, Marlina, S Suresh Kumar
Dengue virus (DENV) has emerged as a major economic concern in developing countries, with 2.5 billion people believed to be at risk. Vascular endothelial cells (ECs) lining the circulatory system from heart to end vessels perform crucial functions in the human body, by aiding gas exchange in lungs, gaseous, nutritional and its waste exchange in all tissues, including the blood brain barrier, filtration of fluid in the glomeruli, neutrophil recruitment, hormone trafficking, as well as maintenance of blood vessel tone and hemostasis...
July 2017: Acta Tropica
Elena Bekerman, Gregory Neveu, Ana Shulla, Jennifer Brannan, Szu-Yuan Pu, Stanley Wang, Fei Xiao, Rina Barouch-Bentov, Russell R Bakken, Roberto Mateo, Jennifer Govero, Claude M Nagamine, Michael S Diamond, Steven De Jonghe, Piet Herdewijn, John M Dye, Glenn Randall, Shirit Einav
Global health is threatened by emerging viral infections, which largely lack effective vaccines or therapies. Targeting host pathways that are exploited by multiple viruses could offer broad-spectrum solutions. We previously reported that AAK1 and GAK, kinase regulators of the host adaptor proteins AP1 and AP2, are essential for hepatitis C virus (HCV) infection, but the underlying mechanism and relevance to other viruses or in vivo infections remained unknown. Here, we have discovered that AP1 and AP2 cotraffic with HCV particles in live cells...
April 3, 2017: Journal of Clinical Investigation
Yuan Tian, Alessandro Sette, Daniela Weiskopf
Dengue virus (DENV) has spread through most tropical and subtropical areas of the world and represents a serious public health problem. The control of DENV infection has not yet been fully successful due to lack of effective therapeutics or vaccines. Nevertheless, a better understanding of the immune responses against DENV infection may reveal new strategies for eliciting and improving antiviral immunity. T cells provide protective immunity against various viral infections by generating effector cells that cooperate to eliminate antigens and memory cells that can survive for long periods with enhanced abilities to control recurring pathogens...
2016: Frontiers in Immunology
Andrea Troupin, Devon Shirley, Berlin Londono-Renteria, Alan M Watson, Cody McHale, Alex Hall, Adam Hartstone-Rose, William B Klimstra, Gregorio Gomez, Tonya M Colpitts
Dengue virus (DENV) is a mosquito-borne flavivirus that causes serious global human disease and mortality. Skin immune cells are an important component of initial DENV infection and systemic spread. Here, we show that mast cells are a target of DENV in human skin and that DENV infection of skin mast cells induces degranulation and alters cytokine and growth factor expression profiles. Importantly, to our knowledge, we also demonstrate for the first time that DENV localizes within secretory granules in infected skin mast cells...
December 1, 2016: Journal of Immunology: Official Journal of the American Association of Immunologists
Stefan W Metz, Shaomin Tian, Gabriel Hoekstra, Xianwen Yi, Michelle Stone, Katie Horvath, Michael J Miley, Joseph DeSimone, Chris J Luft, Aravinda M de Silva
Dengue virus (DENV) is the causative agent of dengue fever and dengue hemorrhagic fever. The virus is endemic in over 120 countries, causing over 350 million infections per year. Dengue vaccine development is challenging because of the need to induce simultaneous protection against four antigenically distinct DENV serotypes and evidence that, under some conditions, vaccination can enhance disease due to specific immunity to the virus. While several live-attenuated tetravalent dengue virus vaccines display partial efficacy, it has been challenging to induce balanced protective immunity to all 4 serotypes...
October 2016: PLoS Neglected Tropical Diseases
Yi-ying Chou, Christian Cuevas, Margot Carocci, Sarah H Stubbs, Minghe Ma, David K Cureton, Luke Chao, Frances Evesson, Kangmin He, Priscilla L Yang, Sean P Whelan, Susan R Ross, Tom Kirchhausen, Raphaël Gaudin
UNLABELLED: Virus entry into cells is a multistep process that often requires the subversion of subcellular machineries. A more complete understanding of these steps is necessary to develop new antiviral strategies. While studying the potential role of the actin network and one of its master regulators, the small GTPase Cdc42, during Junin virus (JUNV) entry, we serendipitously uncovered the small molecule ZCL278, reported to inhibit Cdc42 function as an entry inhibitor for JUNV and for vesicular stomatitis virus, lymphocytic choriomeningitis virus, and dengue virus but not for the nonenveloped poliovirus...
May 2016: Journal of Virology
Valerie A Villareal, Mary A Rodgers, Deirdre A Costello, Priscilla L Yang
Lipids are necessary for every step in the replication cycle of hepatitis C virus (HCV) and dengue virus (DENV), members of the family Flaviviridae. Recent studies have demonstrated that discrete steps in the replication cycles of these viruses can be inhibited by pharmacological agents that target host factors mediating lipid synthesis, metabolism, trafficking, and signal transduction. Despite this, targeting host lipid metabolism and trafficking as an antiviral strategy by blockade of entire pathways may be limited due to host toxicity...
December 2015: Antiviral Research
Luana E Piccini, Viviana Castilla, Elsa B Damonte
The endocytic uptake and intracellular trafficking for penetration of DENV-3 strain H-87 into Vero cells was analyzed by using several biochemical inhibitors and dominant negative mutants of cellular proteins. The results presented show that the infective entry of DENV-3 into Vero cells occurs through a non-classical endocytosis pathway dependent on low pH and dynamin, but non-mediated by clathrin. After uptake, DENV-3 transits through early endosomes to reach Rab 7-regulated late endosomes, and according with the half-time for ammonium chloride resistance viral nucleocapsid is released into the cytosol approximately at 12 min post-infection...
2015: PloS One
Qian Shu, Nicholas J Lennemann, Saumendra N Sarkar, Yoel Sadovsky, Carolyn B Coyne
Interferon stimulated genes (ISGs) target viruses at various stages of their infectious life cycles, including at the earliest stage of viral entry. Here we identify ArfGAP with dual pleckstrin homology (PH) domains 2 (ADAP2) as a gene upregulated by type I IFN treatment in a STAT1-dependent manner. ADAP2 functions as a GTPase-activating protein (GAP) for Arf6 and binds to phosphatidylinositol 3,4,5-trisphosphate (PI(3,4,5)P3) and PI(3,4)P2. We show that overexpression of ADAP2 suppresses dengue virus (DENV) and vesicular stomatitis virus (VSV) infection in an Arf6 GAP activity-dependent manner, while exerting no effect on coxsackievirus B (CVB) or Sendai virus (SeV) replication...
September 2015: PLoS Pathogens
Qiang Fu, Bahar Inankur, John Yin, Rob Striker, Que Lan
Host factors that enable dengue virus (DENV) to propagate in the mosquito host cells are unclear. It is known that cellular cholesterol plays an important role in the life cycle of DENV in human host cells but unknown if the lipid requirements differ for mosquito versus mammalian. In mosquito Aedes aegypti, sterol carrier protein 2 (SCP-2) is critical for cellular cholesterol homeostasis. In this study, we identified SCP-2 as a critical host factor for DENV production in mosquito Aag2 cells. Treatment with a small molecule commonly referred to as SCPI-1, (N-(4-{[4-(3,4-dichlorophenyl)-1,3-thiazol-2-yl]amino}phenyl)acetamide hydrobromide, a known inhibitor of SCP-2, or knockdown of SCP-2 dramatically repressed the virus production in mosquito but not mammalian cells...
September 2015: Journal of Medical Entomology
Philippe Metz, Abhilash Chiramel, Laurent Chatel-Chaix, Gualtiero Alvisi, Peter Bankhead, Rodrigo Mora-Rodriguez, Gang Long, Anne Hamacher-Brady, Nathan R Brady, Ralf Bartenschlager
UNLABELLED: Autophagic flux involves formation of autophagosomes and their degradation by lysosomes. Autophagy can either promote or restrict viral replication. In the case of Dengue virus (DENV), several studies report that autophagy supports the viral replication cycle, and describe an increase of autophagic vesicles (AVs) following infection. However, it is unknown how autophagic flux is altered to result in increased AVs. To address this question and gain insight into the role of autophagy during DENV infection, we established an unbiased, image-based flow cytometry approach to quantify autophagic flux under normal growth conditions and in response to activation by nutrient deprivation or them TOR inhibitor Torin1...
August 2015: Journal of Virology
Ming Yuan Li, Marc Grandadam, Kevin Kwok, Thibault Lagache, Yu Lam Siu, Jing Shu Zhang, Kouxiong Sayteng, Mateusz Kudelko, Cheng Feng Qin, Jean-Christophe Olivo-Marin, Roberto Bruzzone, Pei Gang Wang
Membrane receptors at the surface of target cells are key host factors for virion entry; however, it is unknown whether trafficking and secretion of progeny virus requires host intracellular receptors. In this study, we demonstrate that dengue virus (DENV) interacts with KDEL receptors (KDELR), which cycle between the ER and Golgi apparatus, for vesicular transport from ER to Golgi. Depletion of KDELR by siRNA reduced egress of both DENV progeny and recombinant subviral particles (RSPs). Coimmunoprecipitation of KDELR with dengue structural protein prM required three positively charged residues at the N terminus, whose mutation disrupted protein interaction and inhibited RSP transport from the ER to the Golgi...
March 3, 2015: Cell Reports
Pratap Parida, R N S Yadav, Kishore Sarma
Dengue infections produce a distinct character of virus-induced intracellular membrane alterations which are associated with the viral replication machinery. Currently, the NS3 protein is being targeted for antiviral therapy against dengue. NS3 protein of dengue virus interacts with nuclear receptor binding protein (NRBP) of human causing cell trafficking between the Endoplasmic Reticulum (ER) and Golgi, which interacts with Rac3, a member of the Rho-GTPase family. No crystal structure of the NRBP is available for any species, thus limiting the complete understanding of structure- function relationships of this protein...
2014: Current Pharmaceutical Biotechnology
Wei-Chun Tang, Ren-Jye Lin, Ching-Len Liao, Yi-Ling Lin
UNLABELLED: Positive-sense RNA viruses, such as dengue virus (DENV), hijack the intracellular membrane machinery for their own replication. The Rab18 protein, a member of the Rab GTPase family, key regulators of membrane trafficking, is located on the organelles involved in DENV infection, such as the endoplasmic reticulum (ER) and lipid droplets (LDs). In this study, we addressed the potential involvement of Rab18 in DENV infection by using cells overexpressing the wild-type, GTP-bound active form, or GDP-bound inactive form of Rab18 and cells with Rab18 knockdown...
June 2014: Journal of Virology
Summer Zhang, Kuan Rong Chan, Hwee Cheng Tan, Eng Eong Ooi
The early events of the dengue virus life cycle involve virus binding, internalization, trafficking, and fusion. Fluorescently labeled viruses can be used to visualize these early processes. As dengue virus has 180 identical copies of the envelope protein attached to the membrane surface and is surrounded by a lipid membrane, amine-reactive (Alexa Fluor) or lipophilic (DiD) dyes can be used for virus labeling. These dyes are highly photostable and are ideal for studies involving cellular uptake and endosomal transport...
2014: Methods in Molecular Biology
Hongliang Wang, Jeffrey W Perry, Adam S Lauring, Petra Neddermann, Raffaele De Francesco, Andrew W Tai
BACKGROUND & AIMS: Positive-sense RNA viruses remodel intracellular membranes to generate specialized membrane compartments for viral replication. Several RNA viruses, including poliovirus and hepatitis C virus (HCV), require phosphatidylinositol (PI) 4-kinases for their replication. However, it is not known how PI 4-kinases and their product, PI(4)P, facilitate host membrane reorganization and viral replication. In addition, although the HCV replication compartment, known as the membranous web, is believed to be cholesterol enriched, the mechanisms by which this occurs have not been elucidated...
May 2014: Gastroenterology
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