Inhibition of Mayaro Virus Replication by Prostaglandin A1 in Vero Cells

Intervirology ◽  
2001 ◽  
Vol 44 (6) ◽  
pp. 344-349 ◽  
Author(s):  
F.M. Burlandy ◽  
M.A. Rebello
Keyword(s):  
Virus Replication ◽  
Vero Cells ◽  
Mayaro Virus

shRNA targeting nonstructural protein inhibits the replication of severe fever with thrombocytopenia syndrome virus

2021 ◽  
Author(s):  
Lili Dou ◽  
Xiaoli Tao ◽  
Wei Zhao ◽  
Guofeng Zheng ◽  
Ying Lu ◽  
...  
Keyword(s):  
Tissue Culture ◽  
Western Blotting ◽  
Virus Replication ◽  
Nonstructural Protein ◽  
Vero Cells ◽  
Antiviral Effects ◽  
Infective Dose ◽  
Shrna Plasmid ◽  
Dose Dependent ◽  
Severe Fever

Aim: To explore whether shRNA targeting nonstructural protein (NSs) of severe fever with thrombocytopenia syndrome virus (SFTSV) could inhibit SFTSV replication in Vero cells. Materials & methods: SFTSV used in this experiment was propagated in Vero cells and stored at -20°C. shRNA plasmid against NSs of SFTSV was transfected to Vero cells and infected with SFTSV, after which western blotting and tissue culture infective dose (TCID50) were used to measure the virus titers. Results: shRNA against NSs protein decreased the expression of NSs and inhibited the replication of SFTSV. Conclusion: The constructed SFTSV NSs-shRNA plasmid could inhibit the replication of SFTSV. It was concluded that SFTSV NSs-shRNA could inhibit virus replication for at least 72 h. shRNA-mediated antiviral effects were dose-dependent.

scholarly journals The Major Phosphorylation Sites of the Respiratory Syncytial Virus Phosphoprotein Are Dispensable for Virus Replication In Vitro

2002 ◽  
Vol 76 (21) ◽  
pp. 10776-10784 ◽  
Author(s):  
Bin Lu ◽  
Chien-Hui Ma ◽  
Robert Brazas ◽  
Hong Jin
Keyword(s):  
Respiratory Syncytial Virus ◽  
Virus Replication ◽  
Vero Cells ◽  
Phosphorylation Sites ◽  
N Protein ◽  
P Protein ◽  
Infected Cells ◽  
Syncytial Virus

ABSTRACT The phosphoprotein (P protein) of respiratory syncytial virus (RSV) is a key component of the viral RNA-dependent RNA polymerase complex. The protein is constitutively phosphorylated at the two clusters of serine residues (116, 117, and 119 [116/117/119] and 232 and 237 [232/237]). To examine the role of phosphorylation of the RSV P protein in virus replication, these five serine residues were altered to eliminate their phosphorylation potential, and the mutant proteins were analyzed for their functions with a minigenome assay. The reporter gene expression was reduced by 20% when all five phosphorylation sites were eliminated. Mutants with knockout mutations at two phosphorylation sites (S232A/S237A [PP2]) and at five phosphorylation sites (S116L/S117R/S119L/S232A/S237A [PP5]) were introduced into the infectious RSV A2 strain. Immunoprecipitation of 33Pi-labeled infected cells showed that P protein phosphorylation was reduced by 80% for rA2-PP2 and 95% for rA2-PP5. The interaction between the nucleocapsid (N) protein and P protein was reduced in rA2-PP2- and rA2-PP5-infected cells by 30 and 60%, respectively. Although the two recombinant viruses replicated well in Vero cells, rA2-PP2 and, to a greater extent, rA2-PP5, replicated poorly in HEp-2 cells. Virus budding from the infected HEp-2 cells was affected by dephosphorylation of P protein, because the majority of rA2-PP5 remained cell associated. In addition, rA2-PP5 was also more attenuated than rA2-PP2 in replication in the respiratory tracts of mice and cotton rats. Thus, our data suggest that although the major phosphorylation sites of RSV P protein are dispensable for virus replication in vitro, phosphorylation of P protein is required for efficient virus replication in vitro and in vivo.

scholarly journals Deletion of Nonstructural Proteins NS1 and NS2 from Pneumonia Virus of Mice Attenuates Viral Replication and Reduces Pulmonary Cytokine Expression and Disease

2008 ◽  
Vol 83 (4) ◽  
pp. 1969-1980 ◽  
Author(s):  
Ursula J. Buchholz ◽  
Jerrold M. Ward ◽  
Elaine W. Lamirande ◽  
Britta Heinze ◽  
Christine D. Krempl ◽  
...  
Keyword(s):  
Virus Replication ◽  
Vero Cells ◽  
Type I ◽  
Nonstructural Proteins ◽  
Virus Growth ◽  
Broad Array ◽  
Syncytial Virus ◽  
Inducible Genes ◽  
Overt Disease ◽  
Convenient Model

ABSTRACT Pneumonia virus of mice (PVM) strain 15 causes fatal pneumonia in mice and provides a convenient model for human respiratory syncytial virus pathogenesis and immunobiology. We prepared PVM mutants lacking the genes for nonstructural proteins NS1 and/or NS2. In Vero cells, which lack type I interferon (IFN), deletion of these proteins had no effect on the efficiency of virus growth. In IFN-competent mouse embryo fibroblasts, wild-type (wt) PVM and the ΔNS1 virus grew efficiently and strongly inhibited the IFN response, whereas virus lacking NS2 was highly attenuated and induced high levels of IFN and IFN-inducible genes. In BALB/c mice, intranasal infection with wt PVM caused overt disease that began on day 6 and was lethal by day 9 postinoculation. In comparison, ΔNS1 induced transient, reduced disease, and ΔNS2 and ΔNS12 caused no disease. Thus, NS1 and NS2 are virulence factors, with NS2 being a major antagonist of the type I IFN system. The pulmonary titers of wt PVM and ΔNS1 were high on day 3 and increased further by day 6; in addition, expression of IFN and representative proinflammatory cytokines/chemokines and T lymphocyte-related cytokines was undetectable on day 3 but increased dramatically by day 6 coincident with the onset of disease. The titers of ΔNS2 and ΔNS12 were somewhat lower on day 3 and decreased further by day 6; in addition, these viruses induced a more circumscribed set of cytokines/chemokines (IFN, interleukin-6 [IL-6], and CXCL10) that were detected on day 3 and had largely subsided by day 6. Lung immunohistology revealed abundant PVM-positive pneumocytes and bronchial and bronchiolar epithelial cells in wt PVM- and ΔNS1-infected mice on day 6 compared to few PVM-positive foci with ΔNS2 and ΔNS12. These results indicate that severe PVM disease is associated with high, poorly controlled virus replication driving the expression of high levels of pulmonary IFN and a broad array of cytokines/chemokines. In contrast, in the absence of NS2, there was an early, transient innate response involving moderate levels of IFN, IL-6, and CXCL10 that restricted virus replication and prevented disease.

scholarly journals APOBEC3G-Regulated Host Factors Interfere with Measles Virus Replication: Role of REDD1 and Mammalian TORC1 Inhibition

2018 ◽  
Vol 92 (17) ◽  
Author(s):  
Vishakha Tiwarekar ◽  
Julia Wohlfahrt ◽  
Markus Fehrholz ◽  
Claus-Jürgen Scholz ◽  
Susanne Kneitz ◽  
...  
Keyword(s):  
Host Cell ◽  
Virus Replication ◽  
Peripheral Blood ◽  
Measles Virus ◽  
Human Peripheral Blood ◽  
Vero Cells ◽  
Peripheral Blood Lymphocytes ◽  
Host Factors ◽  
Human Peripheral Blood Lymphocytes

ABSTRACTWe found earlier that ectopic expression of the cytidine deaminase APOBEC3G (A3G) in Vero cells inhibits measles virus (MV), respiratory syncytial virus, and mumps virus, while the mechanism of inhibition remained unclear. A microarray analysis revealed that in A3G-transduced Vero cells, several cellular transcripts were differentially expressed, suggesting that A3G regulates the expression of host factors. One of the most upregulated host cell factors, REDD1 (regulated in development and DNA damage response-1, also called DDIT4), reduced MV replication ∼10-fold upon overexpression in Vero cells. REDD1 is an endogenous inhibitor of mTORC1 (mammalian target of rapamycin complex-1), the central regulator of cellular metabolism. Interestingly, rapamycin reduced the MV replication similarly to REDD1 overexpression, while the combination of both did not lead to further inhibition, suggesting that the same pathway is affected. REDD1 silencing in A3G-expressing Vero cells abolished the inhibitory effect of A3G. In addition, silencing of A3G led to reduced REDD1 expression, confirming that its expression is regulated by A3G. In primary human peripheral blood lymphocytes (PBL), expression of A3G and REDD1 was found to be stimulated by phytohemagglutinin (PHA) and interleukin-2. Small interfering RNA (siRNA)-mediated depletion of A3G in PHA-stimulated PBL reduced REDD1 expression and increased viral titers, which corroborates our findings in Vero cells. Silencing of REDD1 also increased viral titers, confirming the antiviral role of REDD1. Finally, pharmacological inhibition of mTORC1 by rapamycin in PHA-stimulated PBL reduced viral replication to the level found in unstimulated lymphocytes, indicating that mTORC1 activity supports MV replication as a proviral host factor.IMPORTANCEKnowledge about host factors supporting or restricting virus replication is required for a deeper understanding of virus-cell interactions and may eventually provide the basis for therapeutic intervention. This work was undertaken predominantly to explain the mechanism of A3G-mediated inhibition of MV, a negative-strand RNA virus that is not affected by the deaminase activity of A3G acting on single-stranded DNA. We found that A3G regulates the expression of several cellular proteins, which influences the capacity of the host cell to replicate MV. One of these, REDD1, which modulates the cellular metabolism in a central position by regulating the kinase complex mTORC1, was identified as the major cellular factor impairing MV replication. These findings show interesting aspects of the function of A3G and the dependence of the MV replication on the metabolic state of the cell. Interestingly, pharmacological inhibition of mTORC1 can be utilized to inhibit MV replication in Vero cells and primary human peripheral blood lymphocytes.

scholarly journals RNA interference targeting virion core protein ORF095 inhibits Goatpox virus replication in Vero cells

2012 ◽  
Vol 9 (1) ◽  
pp. 48 ◽  
Author(s):  
Zhixun Zhao ◽  
Guohua Wu ◽  
Xueliang Zhu ◽  
Xinmin Yan ◽  
Yongxi Dou ◽  
...  
Keyword(s):  
Rna Interference ◽  
Virus Replication ◽  
Core Protein ◽  
Vero Cells ◽  
Goatpox Virus ◽  
Virion Core

Role of Actin Microfilaments in Canine Distemper Virus Replication in Vero Cells

2004 ◽  
Vol 66 (4) ◽  
pp. 409-415 ◽  
Author(s):  
Hideo KATAYAMA ◽  
Masatoshi HORI ◽  
Koichi SATO ◽  
Masahiro KAJITA ◽  
Hiroshi OZAKI ◽  
...  
Keyword(s):  
Virus Replication ◽  
Canine Distemper Virus ◽  
Vero Cells ◽  
Actin Microfilaments ◽  
Canine Distemper

Ribavirin inhibits Chandipura virus replication in Vero cells

2020 ◽  
Vol 92 (12) ◽  
pp. 2969-2975
Author(s):  
Anukumar Balakrishnan ◽  
Amol B. Mun
Keyword(s):  
Virus Replication ◽  
Vero Cells ◽  
Chandipura Virus
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