scholarly journals Interference with glycosylation of glycoproteins. Inhibition of formation of lipid-linked oligosaccharides in vivo

1979 ◽  
Vol 184 (1) ◽  
pp. 113-123 ◽  
Author(s):  
R Datema ◽  
R T Schwarz
Keyword(s):  
Molecular Weight ◽  
Influenza Virus ◽  
Kinetic Analysis ◽  
Protein Glycosylation ◽  
Protein Inhibition ◽  
Infected Cells ◽  
Lag Period ◽  
High Mannose ◽  
Virus Proteins

Influenza-virus-infected cells were labelled with radioactive sugars and extracted to give fractions containing lipid-linked oligosaccharides and glycoproteins. The oligosaccharides linked to lipid were of the ‘high-mannose’ type and contained glucose. In the glycoprotein fraction, radioactivity was associated with virus proteins and found to occur predominantly in the ‘high-mannose’ type of glycopeptides. In the presence of the inhibitors 2-deoxy-D-glucose, 2-deoxy-2-amino-D-glucose (glucosamine), 2-deoxy-2-fluoro-D-glucose and 2-deoxy-2-fluoro-D-mannose incorporation of radiolabelled sugars into lipid- and protein-linked oligosaccharides was decreased. Kinetic analysis showed that the inhibitors affected first the assembly of lipid-linked oligosaccharides and then protein glycosylation after a lag period. During inhibition by deoxyglucose and the fluoro sugars lipid-linked oligosaccharides were formed that contained oligosaccharides of decreased molecular weight. No such aberrant forms were found during inhibition by glucosamine. In the case of inhibition by deoxyglucose it was shown that the aberrant oligosaccharides were not transferred to protein. Inhibition of formation of lipid-linked oligosaccharides by deoxyglucose and fluoro sugars was antagonized by mannose, in which case oligosaccharides of normal molecular weight were formed. The inhibition by glucosamine was reversed by its removal from the medium. The reversible effects of these inhibitors exemplify their usefulness as tools in the study of glycosylation processes.

Effect of Influenza Virus Proteins Modulate Hemostasis in vitro and in vivo

2003 ◽  
Vol 30 (6) ◽  
pp. 596-602
Author(s):  
I. N. Zhilinskaya ◽  
L. A. Lyapina ◽  
O. I. Kiselev ◽  
I. P. Ashmarin
Keyword(s):  
Influenza Virus ◽  
Virus Proteins

scholarly journals The N-Glycome regulates the endothelial-to-hematopoietic transition

2019 ◽  
Author(s):  
Dionna M. Kasper ◽  
Jared Hintzen ◽  
Yinyu Wu ◽  
Joey J. Ghersi ◽  
Hanna K. Mandl ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Blood System ◽  
Protein Glycosylation ◽  
Specific Expression ◽  
Hematopoietic Stem ◽  
Single Cell Rna Sequencing ◽  
Stem And Progenitor Cells ◽  
Complex Hybrid ◽  
High Mannose

AbstractHematopoietic stem and progenitor cells (HSPCs) that establish and maintain the blood system in adult vertebrates arise from the transdifferentiation of hemogenic endothelial cells (hemECs) during embryogenesis. This endothelial-to-hematopoietic transition (EHT) is tightly regulated, but the mechanisms are poorly understood. Here, we show that microRNA (miR)-223-mediated regulation of N-glycan biosynthesis in endothelial cells (ECs) regulates EHT. Single cell RNA-sequencing revealed that miR-223 is enriched in hemECs and in oligopotent nascent HSPCs. miR-223 restricts the EHT of lymphoid/myeloid lineages by suppressing the expression of mannosyltransferase alg2 and sialyltransferase st3gal2, two enzymes involved in N-linked protein glycosylation. High-throughput glycomics of ECs lacking miR-223 showed a decrease of high mannose versus sialylated complex/hybrid sugars on N-glycoproteins involved in EHT such as the metalloprotease Adam10. Endothelial-specific expression of an N-glycan Adam10 mutant or of the N-glycoenzymes phenocopied the aberrant HSPC production of miR-223 mutants. Thus, the N-glycome plays a previously unappreciated role as an intrinsic regulator of EHT, with specific mannose and sialic acid modifications serving as key endothelial determinants of their hematopoietic fate.One Sentence SummaryThe N-glycan “sugar code” governs the hematopoietic fate of endothelial cells and regulates blood stem cell production in vivo.

scholarly journals A Novel Sulfonated Derivative of β-Cyclodextrin Effectively Inhibits Influenza A Virus Infection in vitro and in vivo

Acta Naturae ◽  
2019 ◽  
Vol 11 (3) ◽  
pp. 20-30 ◽  
Author(s):  
E. P. Goncharova ◽  
Y. A. Kostyro ◽  
A. V. Ivanov ◽  
M. A. Zenkova
Keyword(s):  
Influenza Virus ◽  
Influenza A ◽  
High Efficiency ◽  
Influenza Infection ◽  
Lethal Infection ◽  
Novel Drugs ◽  
Infected Cells ◽  
Low Toxicity

The development of novel drugs against the influenza virus with high efficiency and low toxicity is an urgent and important task. Previous reports have demonstrated that compounds based on sulfo derivatives of oligo- and polysaccharides possess high antiviral activity. In this study, we have examined the ability of a novel sulfonated derivative of -cyclodextrin (KS-6469) to inhibit the influenza virus A/WSN/33 (H1N1) infection in vitro and in vivo. The antiviral potential of KS-6469 against the influenza virus was evaluated in Madin-Darby Canine Kidney epithelial cells treated with serially diluted KS-6469. We found out that KS-6469 completely inhibited viral reproduction after treatment of the infected cells with the compound for 48 h. Our data show that double intranasal treatment of mice with KS-6469 fully protected the animals from a lethal infection and significantly decreased the viral titers in the lungs of the infected animals. Thus, the novel sulfonated -cyclodextrin derivative KS-6469 is a promising candidate for the development of antiviral drugs for preventing and treating the influenza infection.

scholarly journals Localization of influenza virus proteins to nuclear dot 10 structures in influenza virus-infected cells

Virology ◽  
2003 ◽  
Vol 310 (1) ◽  
pp. 29-40 ◽  
Author(s):  
Yoshiko Sato ◽  
Kenichi Yoshioka ◽  
Chie Suzuki ◽  
Satoshi Awashima ◽  
Yasuhiro Hosaka ◽  
...  
Keyword(s):  
Influenza Virus ◽  
Infected Cells ◽  
Virus Proteins

scholarly journals Glycoprotein biosynthesis in quiescent and stimulated thymocytes and a T-cell lymphoma. The synthesis of high-mannose oligosaccharides

1982 ◽  
Vol 201 (2) ◽  
pp. 377-385 ◽  
Author(s):  
C A Rupar ◽  
G M Cook
Keyword(s):  
Molecular Weight ◽  
Cell Lymphoma ◽  
Malignant Cell ◽  
Protein Glycosylation ◽  
Elution Profile ◽  
Malignant Cells ◽  
Cell Type ◽  
Quiescent Cells ◽  
Metabolic Labelling ◽  
High Mannose

Quiescent thymocytes, mitogen-stimulated thymocytes and acute-leukaemic lymphoblasts provide a model for the study of protein glycosylation in quiescent cells, mitotically active non-malignant and malignant cells respectively. The biosynthesis of both complex and high-mannose-type oligosaccharides was monitored by metabolic labelling with [6-3]fucose and [2-3H]mannose. Bio-Gel P6 elution profiles of [6-3H]fucose-labelled glycopeptides showed that quiescent thymocytes and stimulated thymocytes synthesized qualitatively and quantitatively similar glycopeptides; however, higher-molecular-weight glycopeptides were synthesized by the acute-leukaemic lymphoblasts. The amount of [2(-3)H]mannose incorporated into glycopeptide by quiescent thymocytes was less than 10% of that incorporated by stimulated thymocytes. The Bio-Gel P6 elution profile of [2(-3)H]mannose-labelled glycopeptides from acute leukaemic lymphoblasts was qualitatively similar to that of stimulated thymocytes, with about 40% of the radioactivity incorporated into one glycopeptide peak. This glycopeptide was characterized by Bio-Gel P6 and concanavalin A affinity chromatography, radioactive-sugar analysis, sensitivity to alpha-mannosidase and endoglycosidase H and resistance to beta-glucosaminidase as containing a high-mannose oligosaccharide, possible of Man7-8GlcNAc2 structure. Pulse/chase experiments indicated that this high-mannose oligosaccharide was an end product and not a biosynthetic intermediate. It is concluded that higher-molecular-weight fucose-labelled glycopeptides are characteristic of the malignant cell type, and the synthesis of high-mannose oligosaccharide, Man7-8GlcNAc2, in stimulated thymocytes and acute-leukaemic lymphoblasts is associated with mitotically active cells.

scholarly journals Transcriptome Analysis of Infected and Bystander Type 2 Alveolar Epithelial Cells during Influenza A Virus Infection Reveals In Vivo Wnt Pathway Downregulation

2018 ◽  
Vol 92 (21) ◽  
Author(s):  
Aidan S. Hancock ◽  
Christopher J. Stairiker ◽  
Alina C. Boesteanu ◽  
Elisa Monzón-Casanova ◽  
Sebastian Lukasiak ◽  
...  
Keyword(s):  
Influenza Virus ◽  
Wnt Signaling ◽  
Wnt Signaling Pathway ◽  
A549 Cells ◽  
Alveolar Epithelial Cells ◽  
Epithelial Stem Cells ◽  
Alveolar Epithelial ◽  
Lung Epithelial ◽  
Infected Cells

ABSTRACT Influenza virus outbreaks remain a serious threat to public health. A greater understanding of how cells targeted by the virus respond to the infection can provide insight into the pathogenesis of disease. Here we examined the transcriptional profile of in vivo-infected and uninfected type 2 alveolar epithelial cells (AEC) in the lungs of influenza virus-infected mice. We show for the first time the unique gene expression profiles induced by the in vivo infection of AEC as well as the transcriptional response of uninfected bystander cells. This work allows us to distinguish the direct and indirect effects of infection at the cellular level. Transcriptome analysis revealed that although directly infected and bystander AEC from infected animals shared many transcriptome changes compared to AEC from uninfected animals, directly infected cells produce more interferon and express lower levels of Wnt signaling-associated transcripts, while concurrently expressing more transcripts associated with cell death pathways, than bystander uninfected AEC. The Wnt signaling pathway was downregulated in both in vivo-infected AEC and in vitro-infected human lung epithelial A549 cells. Wnt signaling did not affect type I and III interferon production by infected A549 cells. Our results reveal unique transcriptional changes that occur within infected AEC and show that influenza virus downregulates Wnt signaling. In light of recent findings that Wnt signaling is essential for lung epithelial stem cells, our findings reveal a mechanism by which influenza virus may affect host lung repair. IMPORTANCE Influenza virus infection remains a major public health problem. Utilizing a recombinant green fluorescent protein-expressing influenza virus, we compared the in vivo transcriptomes of directly infected and uninfected bystander cells from infected mouse lungs and discovered many pathways uniquely regulated in each population. The Wnt signaling pathway was downregulated in directly infected cells and was shown to affect virus but not interferon production. Our study is the first to discern the in vivo transcriptome changes induced by direct viral infection compared to mere exposure to the lung inflammatory milieu and highlight the downregulation of Wnt signaling. This downregulation has important implications for understanding influenza virus pathogenesis, as Wnt signaling is critical for lung epithelial stem cells and lung epithelial cell differentiation. Our findings reveal a mechanism by which influenza virus may affect host lung repair and suggest interventions that prevent damage or accelerate recovery of the lung.

Influence of the host cell on influenza virus replication

1980 ◽  
Vol 288 (1029) ◽  
pp. 349-357 ◽  
Keyword(s):  
Influenza Virus ◽  
Host Cell ◽  
Messenger Rna ◽  
Actinomycin D ◽  
Late Protein ◽  
Abnormal Accumulation ◽  
Infected Cells ◽  
P Proteins ◽  
Cellular Dna ◽  
Virus Proteins

The replication of influenza virus is characterized by a unique dependence upon host cell nuclear function. In contrast to all other negative strand RNA viruses, transcription from host cellular DNA is a prerequisite for the synthesis of virus-specific messenger RNA; new DNA synthesis is not required. We have analysed the distribution of each of the nine virus-specified proteins between the nucleus and cytoplasm of virus-infected cells, and find that in addition to the NP and the NSj proteins, two of the three P proteins show preferential migration into the nucleus. This subgroup of virus proteins may be involved in the early transcription of the viral genome which probably occurs in the nucleus. In non-permissive cell lines and in cells whose DNA function has been impaired by treatment with ultraviolet light, A-acetoxyacetaminofluorene or low doses of actinomycin D, production of some late virus proteins is inhibited. The specific host function required for this switch to late protein synthesis is unknown but in the cells treated with actinomycin D an abnormal accumulation of virus-specific mRNA occurs in the nucleus. In all cases studied, synthesis of new vRNA ceases when production of these late proteins has been blocked.

scholarly journals Fine specificity of cytotoxic T lymphocytes primed in vivo either with virus or synthetic lipopeptide vaccine or primed in vitro with peptide.

1991 ◽  
Vol 174 (6) ◽  
pp. 1665-1668 ◽  
Author(s):  
H Schild ◽  
M Norda ◽  
K Deres ◽  
K Falk ◽  
O Rötzschke ◽  
...  
Keyword(s):  
Influenza Virus ◽  
T Lymphocytes ◽  
Cytotoxic T Lymphocytes ◽  
Synthetic Peptide ◽  
Fine Specificity ◽  
Peptide Mixtures ◽  
Infected Cells ◽  
Do So

Standard synthetic peptide preparations contain numerous peptidic byproducts in small amounts, which may be efficiently recognized by cytotoxic T lymphocytes (CTL). Recognition patterns of such peptide mixtures by CTL may serve as a kind of fingerprint for CTL fine specificity. Three types of H-2Db-restricted CTL were compared in this way. CTL primed in vivo either with A/PR/8/34 influenza virus or with a synthetic lipopeptide vaccine prepared from influenza nucleoprotein (NP) peptide 365-380 showed identical fine specificity. Both recognize virus-infected cells. In contrast, CTL primed in vitro with NP 365-380 had a different fine specificity and they did not recognize virus-infected cells. Most significantly, the two in vivo primed CTL types efficiently recognized the natural viral nonapeptide NP 366-374 presented by virus-infected H-2b cells, whereas the in vitro primed CTL failed to do so.

scholarly journals Glycoprotein biosynthesis in animal cells grown in suspension culture. Assembly of lipid-linked saccharides and formation of protein-bound ‘high-mannose’ oligosaccharides

1981 ◽  
Vol 195 (1) ◽  
pp. 139-151 ◽  
Author(s):  
D S Bailey ◽  
J Burke ◽  
R Sinclair ◽  
B B Mukherjee
Keyword(s):  
Molecular Weight ◽  
Suspension Culture ◽  
High Molecular Weight ◽  
Deae Cellulose ◽  
Glucose Deprivation ◽  
Animal Cells ◽  
Glycoprotein Biosynthesis ◽  
High Mannose

Glycoprotein biosynthesis was studied with mouse L-cells grown in suspension culture. Glucose-deprived cells incorporated [3H]mannose into ‘high-mannose’ protein-bound oligosaccharides and a few relatively high-molecular-weight lipid-linked oligosaccharides. The latter were retained by DEAE-cellulose and turned over quite slowly during pulse--chase experiments. Increased heterogeneity in size of lipid-linked oligosaccharides developed during prolonged glucose deprivation. Sequential elongation of lipid-linked oligosaccharides was also observed, and conditions that prevented the assembly of the higher lipid-linked oligosaccharides also prevented the formation of the larger protein-bound ‘high-mannose’ oligosaccharides. In parallel experiments, [3H]mannose was incorporated into a total polyribosome fraction, suggesting that mannose residues were transferred co-translationally to nascent protein. Membrane preparations from these cells catalysed the assembly from UDP-N-acetyl-D-[6-3H]glucosamine and GDP-D-[U-14C]mannose of polyisoprenyl diphosphate derivatives whose oligosaccharide moieties were heterogeneous in size. Elongation of the N-acetyl-D-[6-3H]glucosamine-initiated glycolipids with mannose residues produced several higher lipid-linked oligosaccharides similar to those seen during glucose deprivation in vivo. Glucosylation of these mannose-containing oligosaccharides from UDP-D-[6-3H]glucose was restricted to those of a relatively high molecular weight. Protein-bound saccharides formed in vitro were mainly smaller in size than those assembled on the lipid acceptors. These results support the involvement of lipid-linked saccharides in the synthesis of asparagine-linked glycoproteins, but show both in vivo and in vitro that protein-bound ‘high-mannose’ oligosaccharide formation can occur independently of higher lipid-linked oligosaccharide synthesis.

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