Deciphering the Role of Bovine Viral Diarrhea Virus Non-Structural NS4B Protein in Viral Pathogenesis

Bovine viral diarrhea virus (BVDV) is a (+) ssRNA virus that belongs to the family <i>Flaviviridae</i>. BVDV is a significant animal pathogen causing substantial economic losses to the cattle industry worldwide through respiratory and gastrointestinal infections and abortion or birth of...

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Main Authors: Shahbaz Bashir, Andrey Kossarev, Violeta Cascon Martin, Jan Paeshuyse
Format: Article
Language:English
Published: MDPI AG 2020-10-01
Series:Veterinary Sciences
Subjects:
Online Access:https://www.mdpi.com/2306-7381/7/4/169
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author Shahbaz Bashir
Andrey Kossarev
Violeta Cascon Martin
Jan Paeshuyse
author_facet Shahbaz Bashir
Andrey Kossarev
Violeta Cascon Martin
Jan Paeshuyse
author_sort Shahbaz Bashir
collection DOAJ
description Bovine viral diarrhea virus (BVDV) is a (+) ssRNA virus that belongs to the family <i>Flaviviridae</i>. BVDV is a significant animal pathogen causing substantial economic losses to the cattle industry worldwide through respiratory and gastrointestinal infections and abortion or birth of persistently infected calves. While the immunogenic profile of some of the BVDV proteins (i.e., E<sup>rns</sup>, E2 and NS3) is well established during viral pathogenesis, very little information is available about most of BVDV’s non-structural proteins in this regard. In recent times, the NS4B protein has emerged as an interesting target of diagnostic, vaccination and therapeutic value in viral infections of other members of the family <i>Flaviviridae</i> due to its key scaffold-like contribution in the viral replication complex. Although, BVDV-NS4B has a membrane topology alongside its role in induction of autophagosomes <i>in vitro</i>. However, information on its immunogenicity during BVDV pathogenesis and vaccination is scarce. To characterize the immunogenic profile of the NS4B, five cows were vaccinated with the live attenuated BVDV vaccine Bovela<sup>®</sup> and blood samples were taken pre- and post-immunization for serum isolation. Virus neutralization assay (VNA) confirmed the presence of anti-BVDV antibodies in the sera of vaccinated cows. VNA also revealed pre-existing antibodies against BVDV in the pre-immunization sera of two cows. To identify BVDV-NS4B specific antibodies, the NS4B protein was expressed in mammalian cells by using the pCI-neo vector system. The sera from BVDV vaccinated cows were evaluated for the presence of BVDV-NS4B specific antibodies through western blot and indirect ELISA. Interestingly, t sera from cows with pre-existing immunity against BVDV were able to detect NS4B in western blot and ELISA, suggesting the presence of NS4B-specific antibodies. The obtained results provide the first indication of the immunogenic nature of BVDV-NS4B protein in sero-converted animals. These findings are consistent with the observation made for NS4B in other <i>Flaviviridae</i> members and confirm this protein as an interesting target with diagnostic, vaccination and therapeutic value.
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spelling doaj.art-23d17347d46f41699eea4a51da524dec2023-11-20T19:23:39ZengMDPI AGVeterinary Sciences2306-73812020-10-017416910.3390/vetsci7040169Deciphering the Role of Bovine Viral Diarrhea Virus Non-Structural NS4B Protein in Viral PathogenesisShahbaz Bashir0Andrey Kossarev1Violeta Cascon Martin2Jan Paeshuyse3Laboratory of Host Pathogen Interaction in Livestock, Department of Biosystems, Division of Animal and Human Health Engineering, KU Leuven University, 3000 Leuven, BelgiumLaboratory of Host Pathogen Interaction in Livestock, Department of Biosystems, Division of Animal and Human Health Engineering, KU Leuven University, 3000 Leuven, BelgiumLaboratory of Host Pathogen Interaction in Livestock, Department of Biosystems, Division of Animal and Human Health Engineering, KU Leuven University, 3000 Leuven, BelgiumLaboratory of Host Pathogen Interaction in Livestock, Department of Biosystems, Division of Animal and Human Health Engineering, KU Leuven University, 3000 Leuven, BelgiumBovine viral diarrhea virus (BVDV) is a (+) ssRNA virus that belongs to the family <i>Flaviviridae</i>. BVDV is a significant animal pathogen causing substantial economic losses to the cattle industry worldwide through respiratory and gastrointestinal infections and abortion or birth of persistently infected calves. While the immunogenic profile of some of the BVDV proteins (i.e., E<sup>rns</sup>, E2 and NS3) is well established during viral pathogenesis, very little information is available about most of BVDV’s non-structural proteins in this regard. In recent times, the NS4B protein has emerged as an interesting target of diagnostic, vaccination and therapeutic value in viral infections of other members of the family <i>Flaviviridae</i> due to its key scaffold-like contribution in the viral replication complex. Although, BVDV-NS4B has a membrane topology alongside its role in induction of autophagosomes <i>in vitro</i>. However, information on its immunogenicity during BVDV pathogenesis and vaccination is scarce. To characterize the immunogenic profile of the NS4B, five cows were vaccinated with the live attenuated BVDV vaccine Bovela<sup>®</sup> and blood samples were taken pre- and post-immunization for serum isolation. Virus neutralization assay (VNA) confirmed the presence of anti-BVDV antibodies in the sera of vaccinated cows. VNA also revealed pre-existing antibodies against BVDV in the pre-immunization sera of two cows. To identify BVDV-NS4B specific antibodies, the NS4B protein was expressed in mammalian cells by using the pCI-neo vector system. The sera from BVDV vaccinated cows were evaluated for the presence of BVDV-NS4B specific antibodies through western blot and indirect ELISA. Interestingly, t sera from cows with pre-existing immunity against BVDV were able to detect NS4B in western blot and ELISA, suggesting the presence of NS4B-specific antibodies. The obtained results provide the first indication of the immunogenic nature of BVDV-NS4B protein in sero-converted animals. These findings are consistent with the observation made for NS4B in other <i>Flaviviridae</i> members and confirm this protein as an interesting target with diagnostic, vaccination and therapeutic value.https://www.mdpi.com/2306-7381/7/4/169bovine viral diarrhea virusNS4Bimmunogenicantibodieswestern blot
spellingShingle Shahbaz Bashir
Andrey Kossarev
Violeta Cascon Martin
Jan Paeshuyse
Deciphering the Role of Bovine Viral Diarrhea Virus Non-Structural NS4B Protein in Viral Pathogenesis
Veterinary Sciences
bovine viral diarrhea virus
NS4B
immunogenic
antibodies
western blot
title Deciphering the Role of Bovine Viral Diarrhea Virus Non-Structural NS4B Protein in Viral Pathogenesis
title_full Deciphering the Role of Bovine Viral Diarrhea Virus Non-Structural NS4B Protein in Viral Pathogenesis
title_fullStr Deciphering the Role of Bovine Viral Diarrhea Virus Non-Structural NS4B Protein in Viral Pathogenesis
title_full_unstemmed Deciphering the Role of Bovine Viral Diarrhea Virus Non-Structural NS4B Protein in Viral Pathogenesis
title_short Deciphering the Role of Bovine Viral Diarrhea Virus Non-Structural NS4B Protein in Viral Pathogenesis
title_sort deciphering the role of bovine viral diarrhea virus non structural ns4b protein in viral pathogenesis
topic bovine viral diarrhea virus
NS4B
immunogenic
antibodies
western blot
url https://www.mdpi.com/2306-7381/7/4/169
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