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https://www.readbyqxmd.com/read/29057295/multicolor-stimulated-emission-depletion-sted-microscopy-to-generate-high-resolution-images-of-respiratory-syncytial-virus-particles-and-infected-cells
#1
Masfique Mehedi, Margery Smelkinson, Juraj Kabat, Sundar Ganesan, Peter L Collins, Ursula J Buchholz
Human respiratory syncytial virus (RSV) infection in human lung epithelial A549 cells induces filopodia, cellular protrusions consisting of F-actin, that extend to neighboring uninfected cells (Mehedi et al., 2016). High-resolution imaging via stimulated emission depletion (STED) microscopy revealed filamentous RSV particles along these filopodia, suggesting that filopodia facilitate RSV cell-to-cell spread (Mehedi et al., 2016). In this protocol, we describe how to fix, permeabilize, immunostain, and mount RSV-infected A549 cells for STED imaging...
September 5, 2017: Bio-protocol
https://www.readbyqxmd.com/read/28759570/camp-dependent-activation-of-protein-kinase-a-attenuates-respiratory-syncytial-virus-induced-human-airway-epithelial-barrier-disruption
#2
Fariba Rezaee, Terri J Harford, Debra T Linfield, Ghaith Altawallbeh, Ronald J Midura, Andrei I Ivanov, Giovanni Piedimonte
Airway epithelium forms a barrier to the outside world and has a crucial role in susceptibility to viral infections. Cyclic adenosine monophosphate (cAMP) is an important second messenger acting via two intracellular signaling molecules: protein kinase A (PKA) and the guanidine nucleotide exchange factor, Epac. We sought to investigate effects of increased cAMP level on the disruption of model airway epithelial barrier caused by RSV infection and the molecular mechanisms underlying cAMP actions. Human bronchial epithelial cells were infected with RSV-A2 and treated with either cAMP releasing agent, forskolin, or cAMP analogs...
2017: PloS One
https://www.readbyqxmd.com/read/28744815/mammalian-diaphanous-related-formin-1-restricts-early-phases-of-influenza-a-nws-33-virus-h1n1-infection-in-llc-mk2-cells-by-affecting-cytoskeleton-dynamics
#3
Flora De Conto, Alessandra Fazzi, Sergey V Razin, Maria Cristina Arcangeletti, Maria Cristina Medici, Silvana Belletti, Carlo Chezzi, Adriana Calderaro
Viruses depend on cellular machinery to efficiently replicate. The host cytoskeleton is one of the first cellular systems hijacked by viruses in order to ensure their intracellular transport and promote the development of infection. Our previous results demonstrated that stable microfilaments and microtubules interfered with human influenza A/NWS/33 virus (H1N1) infection in semi-permissive LLC-MK2 cells. Although formins play a key role in cytoskeletal remodelling, few studies addressed a possible role of these proteins in development of viral infection...
July 25, 2017: Molecular and Cellular Biochemistry
https://www.readbyqxmd.com/read/28702128/a-novel-host-factor-for-human-respiratory-syncytial-virus
#4
COMMENT
Masfique Mehedi, Peter L Collins, Ursula J Buchholz
Human respiratory syncytial virus (RSV) is the leading viral cause of severe lower respiratory disease in young children worldwide. As part of a genome-wide siRNA screen, we recently discovered that actin-related protein 2 (ARP2) is a host factor in the RSV replication cycle. ARP2 is a major constituent of the ARP2/3 complex, which catalyzes actin polymerization involved in cell morphology and motility. In the course of investigating this finding, we also found that RSV infection of human lung epithelial A459 cells induced filopodia formation and stimulated cell motility...
2017: Communicative & Integrative Biology
https://www.readbyqxmd.com/read/28561831/a-dynamic-cell-entry-pathway-of-respiratory-syncytial-virus-revealed-by-tracking-the-quantum-dot-labeled-single-virus
#5
Lin Ling Zheng, Chun Mei Li, Shu Jun Zhen, Yuan Fang Li, Cheng Zhi Huang
Studying the cell entry pathway at the single-particle level can provide detailed and quantitative information for the dynamic events involved in virus entry. Indeed, the viral entry dynamics cannot be monitored by static staining methods used in cell biology, and thus virus dynamic tracking could be useful in the development of effective antiviral strategies. Therefore, the aim of this work was to use a quantum dot-based single-particle tracking approach to monitor the cell entry behavior of the respiratory syncytial virus (RSV) in living cells...
May 31, 2017: Nanoscale
https://www.readbyqxmd.com/read/28154158/caveolae-provide-a-specialized-membrane-environment-for-respiratory-syncytial-virus-assembly
#6
Alexander Ludwig, Tra Huong Nguyen, Daniel Leong, Laxmi Iyer Ravi, Boon Huan Tan, Sara Sandin, Richard J Sugrue
Respiratory syncytial virus (RSV) is an enveloped virus that assembles into filamentous virus particles on the surface of infected cells. Morphogenesis of RSV is dependent upon cholesterol-rich (lipid raft) membrane microdomains, but the specific role of individual raft molecules in RSV assembly is not well defined. Here, we show that RSV morphogenesis occurs within caveolar membranes and that both caveolin-1 and cavin-1 (also known as PTRF), the two major structural and functional components of caveolae, are actively recruited to and incorporated into the RSV envelope...
March 15, 2017: Journal of Cell Science
https://www.readbyqxmd.com/read/27926942/actin-related-protein-2-arp2-and-virus-induced-filopodia-facilitate-human-respiratory-syncytial-virus-spread
#7
Masfique Mehedi, Thomas McCarty, Scott E Martin, Cyril Le Nouën, Eugen Buehler, Yu-Chi Chen, Margery Smelkinson, Sundar Ganesan, Elizabeth R Fischer, Linda G Brock, Bo Liang, Shirin Munir, Peter L Collins, Ursula J Buchholz
Human respiratory syncytial virus (RSV) is an enveloped RNA virus that is the most important viral cause of acute pediatric lower respiratory tract illness worldwide, and lacks a vaccine or effective antiviral drug. The involvement of host factors in the RSV replicative cycle remains poorly characterized. A genome-wide siRNA screen in human lung epithelial A549 cells identified actin-related protein 2 (ARP2) as a host factor involved in RSV infection. ARP2 knockdown did not reduce RSV entry, and did not markedly reduce gene expression during the first 24 hr of infection, but decreased viral gene expression thereafter, an effect that appeared to be due to inhibition of viral spread to neighboring cells...
December 2016: PLoS Pathogens
https://www.readbyqxmd.com/read/27670781/host-cytoskeleton-in-respiratory-syncytial-virus-assembly-and-budding
#8
Shadi Shahriari, James Gordon, Reena Ghildyal
Respiratory syncytial virus (RSV) is one of the major pathogens responsible for lower respiratory tract infections (LRTI) in young children, the elderly, and the immunosuppressed. Currently, there are no antiviral drugs or vaccines available that effectively target RSV infections, proving a significant challenge in regards to prevention and treatment. An in-depth understanding of the host-virus interactions that underlie assembly and budding would inform new targets for antiviral development.Current research suggests that the polymerised form of actin, the filamentous or F-actin, plays a role in RSV assembly and budding...
September 26, 2016: Virology Journal
https://www.readbyqxmd.com/read/27464695/a-proteomic-based-workflow-using-purified-respiratory-syncytial-virus-particles-to-identify-cellular-factors-as-drug-targets
#9
Tra Nguyen Huong, Boon Huan Tan, Richard J Sugrue
The identification of cellular factors that play a role in respiratory syncytial virus (RSV) replication is an alternative strategy in the identification of druggable cellular protein that are essential for RSV replication. In this regard experimental strategies that are able to screen relevant proteins from the vast array of proteins in the cellular milieu will facilitate the identification of potential drug targets. In this chapter we describe a procedure where RSV particles are purified from cells that are permissive for RSV infection, and the protein composition of the purified virus particles characterized using a proteomics-based strategy...
2016: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27004689/actin-and-clathrin-dependent-mechanisms-regulate-interferon-gamma-release-after-stimulation-of-human-immune-cells-with-respiratory-syncytial-virus
#10
Jop Jans, Hicham elMoussaoui, Ronald de Groot, Marien I de Jonge, Gerben Ferwerda
BACKGROUND: Respiratory syncytial virus (RSV) can cause recurrent and severe respiratory tract infections. Cytoskeletal proteins are often involved during viral infections, either for cell entry or the initiation of the immune response. The importance of actin and clathrin dynamics for cell entry and the initiation of the cellular immune response against RSV in human immune cells is not known yet. The aim of this study was to investigate the role of actin and clathrin on cell entry of RSV and the subsequent effect on T cell activation and interferon gamma release in human immune cells...
March 22, 2016: Virology Journal
https://www.readbyqxmd.com/read/26909955/effect-of-co-culturing-human-primary-basic-fibroblasts-with-respiratory-syncytial-virus-infected-16-hbe-cells
#11
J-H Wang, L-H Sun, S-K Huang, A-H Chen
This study investigated the effects induced by co-culturing human primary basic fibroblasts (HPBFs) with 16-human bronchial epithelial cells (16-HBE) infected with respiratory syncytial virus (RSV), in particular the transformation of HPBFs into myofibroblasts and secretion of extracellular matrix proteins. HPBFs were co-cultured with 16-HBE cells infected with RSV and quantitatively analyzed. We constructed models of HPBFs co-cultured with 16-HBE cells that were either uninfected (control group) or infected with RSV (experimental group)...
February 5, 2016: Genetics and Molecular Research: GMR
https://www.readbyqxmd.com/read/25926640/the-nectin-4-afadin-protein-complex-and-intercellular-membrane-pores-contribute-to-rapid-spread-of-measles-virus-in-primary-human-airway-epithelia
#12
Brajesh K Singh, Andrew L Hornick, Sateesh Krishnamurthy, Anna C Locke, Crystal A Mendoza, Mathieu Mateo, Catherine L Miller-Hunt, Roberto Cattaneo, Patrick L Sinn
UNLABELLED: The discovery that measles virus (MV) uses the adherens junction protein nectin-4 as its epithelial receptor provides a new vantage point from which to characterize its rapid spread in the airway epithelium. We show here that in well-differentiated primary cultures of airway epithelial cells from human donors (HAE), MV infectious centers form rapidly and become larger than those of other respiratory pathogens: human respiratory syncytial virus, parainfluenza virus 5, and Sendai virus...
July 2015: Journal of Virology
https://www.readbyqxmd.com/read/25556234/new-host-factors-important-for-respiratory-syncytial-virus-rsv-replication-revealed-by-a-novel-microfluidics-screen-for-interactors-of-matrix-m-protein
#13
Sarit Kipper, Samar Hamad, Leon Caly, Dorit Avrahami, Eran Bacharach, David A Jans, Doron Gerber, Monika Bajorek
Although human respiratory syncytial virus (RSV) is the most common cause of bronchiolitis and pneumonia in infants and elderly worldwide, there is no licensed RSV vaccine or effective drug treatment available. The RSV Matrix protein plays key roles in virus life cycle, being found in the nucleus early in infection in a transcriptional inhibitory role, and later localizing in viral inclusion bodies before coordinating viral assembly and budding at the plasma membrane. In this study, we used a novel, high throughput microfluidics platform and custom human open reading frame library to identify novel host cell binding partners of RSV matrix...
March 2015: Molecular & Cellular Proteomics: MCP
https://www.readbyqxmd.com/read/25408253/imaging-analysis-of-human-metapneumovirus-infected-cells-provides-evidence-for-the-involvement-of-f-actin-and-the-raft-lipid-microdomains-in-virus-morphogenesis
#14
Muhammad Raihan Jumat, Tra Nguyen Huong, Puisan Wong, Liat Hui Loo, Boon Huan Tan, Fiona Fenwick, Geoffrey L Toms, Richard J Sugrue
BACKGOUND: Due to difficulties of culturing Human metapneumovirus (HMPV) much of the current understanding of HMPV replication can be inferred from other closely related viruses. The slow rates of virus replication prevent many biochemical analyses of HMPV particles. In this study imaging was used to examine the process of HMPV morphogenesis in individually infected LLC-MK2 cells, and to better characterise the sites of HMPV assembly. This strategy has circumvented the problems associated with slow replication rates and allowed us to characterise both the HMPV particles and the sites of HMPV morphogenesis...
2014: Virology Journal
https://www.readbyqxmd.com/read/25379375/fungal-colonization-with-pneumocystis-correlates-to-increasing-chloride-channel-accessory-1-hclca1-suggesting-a-pathway-for-up-regulation-of-airway-mucus-responses-in-infant-lungs
#15
Francisco J Pérez, Carolina A Ponce, Diego A Rojas, Pablo A Iturra, Rebeca I Bustamante, Myriam Gallo, Karime Hananias, Sergio L Vargas
Fungal colonization with Pneumocystis is associated with increased airway mucus in infants during their primary Pneumocystis infection, and to severity of COPD in adults. The pathogenic mechanisms are under investigation. Interestingly, increased levels of hCLCA1 - a member of the calcium-sensitive chloride conductance family of proteins that drives mucus hypersecretion - have been associated with increased mucus production in patients diagnosed with COPD and in immunocompetent rodents with Pneumocystis infection...
2014: Results in Immunology
https://www.readbyqxmd.com/read/24777856/dngr-1-is-dispensable-for-cd8-t-cell-priming-during-respiratory-syncytial-virus-infection
#16
Lydia R Durant, Catherine Pereira, Aime Boakye, Spyridon Makris, Fahima Kausar, Michelle Goritzka, Cecilia Johansson
During respiratory syncytial virus (RSV) infection CD8(+) T cells both assist in viral clearance and contribute to immunopathology. CD8(+) T cells recognize viral peptides presented by dendritic cells (DCs), which can directly present viral antigens when infected or, alternatively, "cross-present" antigens after endocytosis of dead or dying infected cells. Mouse CD8α(+) and CD103(+) DCs excel at cross-presentation, in part because they express the receptor DNGR-1 that detects dead cells by binding to exposed F-actin and routes internalized cell debris into the cross-presentation pathway...
August 2014: European Journal of Immunology
https://www.readbyqxmd.com/read/24603118/resveratrol-inhibits-phenotypic-switching-of-neointimal-vascular-smooth-muscle-cells-after-balloon-injury-through-blockade-of-notch-pathway
#17
Jing Zhang, Jing Chen, Changwu Xu, Jian Yang, Qing Guo, Qi Hu, Hong Jiang
BACKGROUND: Phenotypic switching of vascular smooth muscle cells (VSMCs) plays an initial role in neointimal hyperplasia, the main cause of many occlusive vascular diseases. The aim of this study was to measure the effects of resveratrol (RSV) on the phenotypic transformation of VSMCs and to investigate its mechanism of action. METHODS: Cultured VSMCs isolated from rat thoracic aorta were prepared with serum starvation for 72 hours followed by RSV treatment (50-200 μmol/L) and 10% serum stimulation...
March 2014: Journal of Cardiovascular Pharmacology
https://www.readbyqxmd.com/read/24462457/myosin-1e-is-a-component-of-the-invadosome-core-that-contributes-to-regulation-of-invadosome-dynamics
#18
Jessica L Ouderkirk, Mira Krendel
Myosin 1e (myo1e) is an actin-based motor protein that has been implicated in cell adhesion and migration. We examined the role of myo1e in invadosomes, actin-rich adhesion structures that are important for degradation and invasion of the extracellular matrix. RSV-transformed BHK-21 cells, which readily form invadosomes and invadosome rosettes, were used as the experimental model. Myo1e localization to the actin-rich core of invadosomes required the proline-rich Tail Homology 2 (TH2) domain. During invadosome rosette expansion, we observed myo1e recruitment to newly forming invadosomes via Tail Homology 1 (TH1)-dependent interactions with the plasma membrane, where it preceded actin and paxillin...
April 1, 2014: Experimental Cell Research
https://www.readbyqxmd.com/read/24390331/rice-stripe-tenuivirus-nsvc2-glycoproteins-targeted-to-the-golgi-body-by-the-n-terminal-transmembrane-domain-and-adjacent-cytosolic-24-amino-acids-via-the-cop-i-and-cop-ii-dependent-secretion-pathway
#19
Min Yao, Xiaofan Liu, Shuo Li, Yi Xu, Yijun Zhou, Xueping Zhou, Xiaorong Tao
UNLABELLED: The NSvc2 glycoproteins encoded by Rice stripe tenuivirus (RSV) share many characteristics common to the glycoproteins found among Bunyaviridae. Within this viral family, glycoproteins targeting to the Golgi apparatus play a pivotal role in the maturation of the enveloped spherical particles. RSV particles, however, adopt a long filamentous morphology. Recently, RSV NSvc2 glycoproteins were shown to localize exclusively to the ER in Sf9 insect cells. Here, we demonstrate that the amino-terminal NSvc2 (NSvc2-N) targets to the Golgi apparatus in Nicotiana benthamiana cells, whereas the carboxyl-terminal NSvc2 (NSvc2-C) accumulates in the endoplasmic reticulum (ER)...
March 2014: Journal of Virology
https://www.readbyqxmd.com/read/24188202/respiratory-syncytial-virus-induces-leukotriene-c4-synthase-expression-in-bronchial-epithelial-cells
#20
Li-hong Sun, Ai-huan Chen, Zi-feng Yang, Jason J Chen, Wen-da Guan, Jie-ling Wu, Sheng Qin, Nan-shan Zhong
BACKGROUND AND OBJECTIVE: Respiratory syncytial virus (RSV) results in acute wheezing in infants and is frequently associated with recurrent wheezing. Although RSV-induced wheezing clinically resembles that of asthma, corticosteroids are not equivalently effective in RSV-associated wheezing. The study sought to determine the mechanisms of RSV-induced wheezing by establishing an in vitro model of RSV-infected human bronchial epithelial cells (16-HBEC). METHODS: Leukotriene C4 synthase (LTC4 S) messenger RNA (mRNA) expression in 16-HBEC was detected using fluorescence quantitative polymerase chain reaction, and the relative level of LTC4 S mRNA was expressed as quotient cycle threshold (qCt) based on the threshold cycle number value compared with that of β-actin...
November 2013: Respirology: Official Journal of the Asian Pacific Society of Respirology
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