scholarly journals Developing an Antiviral Drug Screening System for Anti-Bovine Viral Diarrhea Virus (BVDV) Therapies

2019 ◽  
Vol 69 (4) ◽  
pp. 1255
Author(s):  
A. Mokhtari ◽  
M. Mahzounieh ◽  
O. Madadgar ◽  
A. Ghalyanchi Langeroudi2
Keyword(s):  
Cell Line ◽  
Drug Screening ◽  
Bovine Viral Diarrhea Virus ◽  
Antiviral Drug ◽  
Bovine Viral Diarrhea ◽  
Screening System ◽  
Egfp Gene ◽  
Diarrhea Virus ◽  
Viral Diarrhea ◽  
Viral Diarrhea Virus

Bovine viral diarrhea virus (BVDV) is an economically important animal pathogen affecting cattle. Despite the use of vaccination, test and slaughter practices, BVD remains a serious problem of cattle breeding. This study was conducted in order to develop a cell line that expresses some of BVDV sub-replicons. BVDV-NADL NS3 and 5’UTR were cloned in pWPI-linker B lentiviral plasmid at the upstream of EGFP gene. Consequently, lentiviral vectors containing BVDV-NS3 and BVDV-5’UTR were produced by using the second-generation lentiviral packaging system. By these lentivectors, MDBK cells expressing BVDV-5’UTR and BVDV-NS3 partial fragments were prepared. The efficiency of the infection was evaluated by fluorescence microscopy, western blotting, and RT-PCR. The results indicated that the development of MDBK cell line expressing these transgenes provides a very sensitive antiviral drug screening system for anti-bovine viral diarrhea virus (BVDV) therapies.

 Evaluating large spontaneous deletions in a bovine cell line selected for bovine viral diarrhea virus resistance - MDBK reads mapping to regions in genome  not shared with CRIB cell line v1

2021 ◽  
Author(s):  
aspen.workman not provided ◽  
mike.heaton not provided ◽  
Dennis A. Webster ◽  
Gregory P Harhay ◽  
Tim Smith ◽  
...  
Keyword(s):  
Cell Line ◽  
Bovine Viral Diarrhea Virus ◽  
Virus Entry ◽  
Mdbk Cell ◽  
Genomic Variation ◽  
Host Factors ◽  
Bovine Viral Diarrhea ◽  
Diarrhea Virus ◽  
Viral Diarrhea ◽  
Viral Diarrhea Virus

Bovine viral diarrhea virus (BVDV) entry into bovine cells involves attachment of virions to cellular receptors, internalization, and pH-dependent fusion with endosomal membranes. The primary host receptor for BVDV is CD46; however, the complete set of host factors required for virus entry is unknown. The Madin-Darby bovine kidney (MDBK) cell line is susceptible to BVDV infection, while a derivative cell line (CRIB) is resistant at the level of virus entry. We performed complete genome sequencing of each to identify genomic variation underlying the resistant phenotype with the aim of identifying host factors essential for BVDV entry. Three large compound deletions in the BVDV-resistant CRIB cell line were identified and predicted to disrupt the function or expression of the genesPTPN12,GRID2, andRABGAP1L. However, CRISPR/Cas9 mediated knockout of these genes, individually or in combination, in the parental MDBK cell line did not impact virus entry or replication. Therefore, resistance to BVDV in the CRIB cell line is not due to the apparent spontaneous loss ofPTPN12,GRID2, orRABGAP1Lgene function. Identifying the functional cause of BVDV resistance in the CRIB cell line may require more detailed comparisons of the genomes and epigenomes.

scholarly journals Complete Genome Sequence of Noncytopathic Bovine Viral Diarrhea Virus 1 Contaminating a High-Passage RK-13 Cell Line

2015 ◽  
Vol 3 (5) ◽  
Author(s):  
Bora Nam ◽  
Ganwu Li ◽  
Ying Zheng ◽  
Jianqiang Zhang ◽  
Kathleen M. Shuck ◽  
...  
Keyword(s):  
Cell Line ◽  
Genome Sequence ◽  
Bovine Viral Diarrhea Virus ◽  
Complete Genome Sequence ◽  
Complete Genome ◽  
Bovine Viral Diarrhea ◽  
High Passage ◽  
Diarrhea Virus ◽  
Viral Diarrhea ◽  
Viral Diarrhea Virus

A high-passage rabbit kidney RK-13 cell line (HP-RK-13[KY], originally derived from the ATCC CCL-37 cell line) used in certain laboratories worldwide is contaminated with noncytopathic bovine viral diarrhea virus (ncpBVDV). On complete genome sequence analysis, the virus strain was found to belong to BVDV group 1b.

scholarly journals Elimination of Non-cytopathic Bovine Viral Diarrhea Virus From the LFBK-αvβ6 Cell Line

2021 ◽  
Vol 8 ◽  
Author(s):  
Ashley R. Gray ◽  
Britta A. Wood ◽  
Elisabeth Henry ◽  
Donald P. King ◽  
Valérie Mioulet
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Bovine Viral Diarrhea Virus ◽  
Bovine Viral Diarrhea ◽  
Analytical Sensitivity ◽  
Diarrhea Virus ◽  
Viral Diarrhea ◽  
Mouth Disease Virus ◽  
Viral Diarrhea Virus

The LFBK-αvβ6 cell line is highly sensitive for the isolation of foot-and-mouth disease virus (FMDV) and porcinophilic vesicular viruses. However, LFBK-αvβ6 cells are contaminated with a non-cytopathic bovine viral diarrhea virus (BVDV), which complicates handling procedures in areas where other cell lines are maintained, as well downstream use of viral isolates. In this study, we used an aromatic cationic compound (DB772) to treat LFBK-αvβ6 cells using an approach that has been previously used to eliminate persistent BVDV from fetal fibroblast cell lines. After three cell passages with 4 μM DB772, BVDV could no longer be detected in unclarified cell suspensions using a pan-pestivirus real-time RT-PCR assay, and remained undetectable after treatment was stopped (nine passages) for an additional 28 passages. The analytical sensitivity of the DB772-treated LFBK-αvβ6 cultures (renamed WRL-LFBK-αvβ6) to titrations of FMDV and other vesicular virus isolates was comparable to untreated LFBK-αvβ6 cells. These new BVDV-free cells can be handled without the risk of cross-contaminating other cells lines or reagents, and used for routine diagnostics, in vivo studies and/or preparation of new vaccine strains.

scholarly journals Expression of Bovine Viral Diarrhea Virus Glycoprotein E2 as a Soluble Secreted Form in a Mammalian Cell Line

2006 ◽  
Vol 13 (6) ◽  
pp. 698-701 ◽  
Author(s):  
Gaetano Donofrio ◽  
Ezio Bottarelli ◽  
Cavirani Sandro ◽  
Cesidio Filippo Flammini
Keyword(s):  
Cell Line ◽  
Bovine Viral Diarrhea Virus ◽  
Natural Infection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Bovine Viral Diarrhea ◽  
Type I ◽  
Diarrhea Virus ◽  
Viral Diarrhea ◽  
Glycoprotein E2 ◽  
Viral Diarrhea Virus

ABSTRACT Bovine viral diarrhea virus (BVDV) membrane-anchored type I glycoprotein E2 is an ∼53-kDa immunodominant glycoprotein inducing the production of neutralizing antibodies in the animal host after natural infection or following immunization with live or killed vaccines. The E2 coding region lacking the transmembrane domain was constructed in a soluble secreted form (secE2) and expressed in the medium of a transiently transfected human cell line. The crude conditioned medium containing secE2 can be potentially employed to develop an enzyme-linked immunosorbent assay antigen for the diagnosis of BVDV infection or for vaccine purposes.

scholarly journals Effects of Interferon, Ribavirin, and Iminosugar Derivatives on Cells Persistently Infected with Noncytopathic Bovine Viral Diarrhea Virus

2004 ◽  
Vol 48 (2) ◽  
pp. 497-504 ◽  
Author(s):  
David Durantel ◽  
Sandra Carrouée-Durantel ◽  
Norica Branza-Nichita ◽  
Raymond A. Dwek ◽  
Nicole Zitzmann
Keyword(s):  
Cell Line ◽  
Bovine Viral Diarrhea Virus ◽  
Alkyl Chain ◽  
Bovine Viral Diarrhea ◽  
Persistently Infected ◽  
Diarrhea Virus ◽  
Viral Diarrhea ◽  
Viral Diarrhea Virus ◽  
Long Alkyl Chain

ABSTRACT Persistent infection with hepatitis C virus (HCV) is a major cause of chronic hepatitis in humans. In chronic carriers, the viral infection induces liver damage that predisposes the patient for cirrhosis and can lead to hepatocellular carcinoma. Current chemotherapies are limited to alpha interferon (IFN-α) used either alone or in combination with ribavirin (RBV). In addition to its limited efficacy, this treatment is frequently poorly tolerated because of its side effects. The urgently needed development of new drugs is made difficult by the lack of an in vitro or in vivo infectivity model, and no cell line has been found so far to reliably and reproducibly support HCV infection. For this reason, the closely related pestivirus bovine viral diarrhea virus (BVDV) has sometimes been used as a surrogate in vitro infectivity model. In this study we used an MDBK cell line persistently infected with noncytopathic BVDV to assess the antiviral effect of IFN-α and RBV, the two drugs currently in clinical use against HCV. The same system was then used to evaluate the potential of two classes of iminosugar derivates to clear noncytopathic BVDV infection from MDBK cells. We show that treatment with long-alkyl-chain deoxynojirimycin derivatives, which are inhibitors of the endoplasmic reticulum (ER)-resident α-glucosidases, can greatly reduce the amount of secreted enveloped viral RNA. Long-alkyl-chain deoxygalactonojirimycin derivatives, which do not inhibit ER α-glucosidases, were less potent but still more effective in this system than IFN-α or ribavirin.

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