Immunofluorescence Studies of Biotype-Specific Expression of Bovine Viral Diarrhoea Virus Epitopes in Infected Cells

Greiser‐Wilke, I.; Dittmar, Kurt E.J.; Ließ, B.; Moennig, V.

doi:10.1099/0022-1317-72-8-2015

Cited by 19 publications

(13 citation statements)

References 18 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Electron microscopy of infected cells has suggested that particles bud into the lumen of modified endoplasmic reticulum structures (15). This hypothesis is also supported by the absence of viral envelope protein expression on the cell surface (16,17,41). In pestivirus virions, the envelope proteins form E2 homodimers and E1-E2 heterodimers (40).…”

Section: The Genus Pestivirus Includes the Important Animal Pathogensmentioning

confidence: 70%

Uncleaved NS2-3 Is Required for Production of Infectious Bovine Viral Diarrhea Virus

Agapov

Murray

Frolov

et al. 2004

J Virol

View full text Add to dashboard Cite

Despite increasing characterization of pestivirus-encoded proteins, functions for nonstructural (NS) proteins NS2, NS2-3, NS4B, and NS5A have not yet been reported. Here we investigated the function of bovine viral diarrhea virus (BVDV) uncleaved NS2-3. To test whether NS2-3 has a discrete function, the uncleaved protein was specifically abolished in two ways: first by inserting a ubiquitin monomer between NS2 and NS3, and second by placing an internal ribosome entry site between the two proteins (a bicistronic genome). In both cases, complete processing of NS2-3 prevented infectious virion formation without affecting RNA replication. We tested the hypothesis that uncleaved NS2-3 was involved in morphogenesis by creating a bicistronic genome in which NS2-3 was restored in the second cistron. With this genome, both uncleaved NS2-3 expression and particle production returned. We then investigated the minimal regions of the polyprotein that could rescue an NS2-3 defect by developing a trans-complementation assay. We determined that the expression of NS4A in cis with NS2-3 markedly increased its activity, while p7 could be supplied in trans. Based on these data, we propose a model for NS2-3 action in virion morphogenesis. The genus Pestivirus includes the important animal pathogens Classical swine fever virus, Border disease virus, and Bovine viral diarrhea virus (BVDV). BVDV infection of cattle can result in asymptomatic infection and seroconversion or a variety of pathologies including fatal mucosal disease (reviewed in reference 29). The genome organization and translation initiation of the pestiviruses closely resemble those of hepatitis C virus. Hepatitis C virus is the sole member of the Hepacivirus genus, which, along with the flaviviruses and pestiviruses, makes up the family Flaviviridae. Because of its close relation to hepatitis C virus, BVDV is increasingly being studied as a surrogate for this important human pathogen (reviewed in reference 23).BVDV is a single-stranded RNA virus of positive polarity with a nonsegmented genome of approximately 12.3 kb. The genome is not capped, and on entry into the cell, translation is initiated at an internal ribosome entry site (IRES) formed by the highly conserved 5Ј nontranslated region (NTR). A single polyprotein is produced and is co-and posttranslationally processed by viral and cellular enzymes. With the exception of the nonvirion protein N pro , the structural proteins are found at the amino terminus of the polyprotein, while the nonstructural (NS) proteins make up the carboxy-terminal portion. The viral proteins are sequentially designated N pro -C-E rns -E1-E2-p7-NS2-NS3-NS4A-NS4B-NS5A-NS5B. N pro is an autoprotease that catalyzes the cleavage after its own carboxy terminus. C is the highly basic nucleocapsid protein and is followed by the viral envelope proteins E rns , E1, and E2. The NS proteins are thought to be components of the membrane-associated viral replicase complex. NS3 has protease, helicase, and NTPase activities and is responsible for the clea...

show abstract

Section: The Genus Pestivirus Includes the Important Animal Pathogensmentioning

confidence: 70%

Uncleaved NS2-3 Is Required for Production of Infectious Bovine Viral Diarrhea Virus

Agapov

Murray

Frolov

et al. 2004

J Virol

View full text Add to dashboard Cite

show abstract

“…Additionally, all proteins could be detected on the cell surface except for E1-MAT. This was totally unexpected for the native proteins, as in cells infected with the wild-type virus all proteins are exclusively localized intracellularly (Greiser-Wilke et al, 1991;Grummer et al, 2001). However, a similar phenomenon has been reported for HCV E1 and E2 proteins in transfected cells, as well as for proteins of plant cells, and is most likely due to saturation/leakiness of ER retention following overexpression (Bartosch et al, 2003;Crofts et al, 1999;Hsu et al, 2003).…”

Section: Discussionmentioning

confidence: 81%

“…E rns and E2 form disulphide-linked homodimers, whereas E1 is found as heterodimers in association with E2 (Rümenapf et al, 1993;Weiland et al, 1990). Neither E rns nor E2 is detectable on the cell surface of infected cells, but they are associated with intracellular membranes (Greiser-Wilke et al, 1991;Grummer et al, 2001). Recently, an endoplasmic reticulum (ER) retention signal within the membrane anchor of the E2 protein was identified (Köhl et al, 2004).…”

Section: Introductionmentioning

confidence: 99%

Formation of bovine viral diarrhea virus E1–E2 heterodimers is essential for virus entry and depends on charged residues in the transmembrane domains

Ronecker

Zimmer

Herrler

et al. 2008

Journal of General Virology

View full text Add to dashboard Cite

The envelope of bovine viral diarrhea virus (BVDV) contains the glycoproteins E rns , E1 and E2. Complementation of a recombinant vesicular stomatitis virus (VSV) with BVDV glycoproteins resulted in infectious pseudotyped viruses. To elucidate the specific role of each of the single envelope glycoproteins during viral entry, pseudotypes were generated bearing the BVDV envelope proteins in different combinations. Pseudoviruses that contained E1 and E2 but not E rns were infectious, indicating that E rns is dispensable for virus entry. VSV/BVDV pseudotypes with chimeric proteins (the ectodomain of the BVDV glycoprotein and the transmembrane domain of the VSV-G protein) were not infectious. The fact that E1-E2 heterodimers were not detected if one of the proteins was chimeric indicated that the heterodimers are crucial for BVDV entry. It was shown by site-directed mutagenesis that the charged amino acids in the transmembrane domains of BVDV E1 (lysine and arginine) and the charged amino acid in the transmembrane domain of E2 (arginine) play a key role in heterodimer formation. Pseudoviruses bearing the mutation E2-R/A, where the charged amino acid was substituted by alanine, were not infectious, supporting the hypothesis that E1-E2 heterodimers are essential for BVDV entry.

show abstract

“…A total of fourteen calves were assigned to the BVDV group and were infected by intranasal inoculation with 10 ml (5 ml per nostril) containing 10 5 50% tissue culture–infective dose (TCID 50 )/ml of ncp BVDV 7443 (subtype 1a), courtesy of the Institut für Virologie TiHo (Hannover, Germany). Ncp BVDV‐1 7443 is a field strain of low virulence isolated from a persistently infected calf (McClurkin et al., ), which has been extensively used in previous in vitro and in vivo studies and that gives reproducible infection profiles (Bolin et al., ; Greiser‐Wilke et al., ; Grummer et al., , ; Pedrera et al., ). Four calves were housed separately as the control group and received intranasally an identical amount of virus‐free cell culture fluid.…”

Section: Methodsmentioning

confidence: 99%

Cell-Mediated Immune Response During Experimental Acute Infection with Bovine Viral Diarrhoea Virus: Evaluation of Blood Parameters

Molina

Risalde

Sánchez-Cordón

et al. 2012

Transbound Emerg Dis

View full text Add to dashboard Cite

Acute infections with bovine viral diarrhoea virus (BVDV), a major pathogen of cattle, are often asymptomatic or produce only mild clinical symptoms. However, they may play an important role in the bovine respiratory disease complex by exerting a marked immunosuppressive effect, as a result of the death of the immunocompetent cell populations involved in controlling innate and adaptive immune responses, together with a marked reduction of both cytokine expression and co-stimulatory molecule synthesis. Although experimental research and field studies have shown that acute BVDV infection enhances susceptibility to secondary infection, the precise mechanism involved in BVDV-induced immunosuppression remains unclear. The present study is aimed at measuring a range of blood parameters in a single group of fourteen calves infected with non-cytopathic BVDV-1. Focus has been put on those related to the cell-mediated immune response just as leucocyte populations and lymphocyte subpopulations, serum concentrations of cytokines (IL-1b, TNF-a, IFN-c, IL-12, IL-4 and IL-10) and acute phase proteins [haptoglobin, serum amyloid A (SAA), fibrinogen and albumin], as well as BVDV-specific antibodies and viremia. After non-cytopathic BVDV-1 infection, clinical signs intensity was never more than moderate coinciding with the presence of viremia and leucocyte and lymphocyte depletion. An early increase in TNF-a, IFN-c and IL-12 levels in contrast to IL-1b was observed in line with a raise in haptoglobin and SAA levels on the latest days of the study. As regards IL-4 levels, no evidence was found of any changes. However, a slight increase in IL-10 was observed, matching up the TNF-a decline during the acute phase response. These findings would help to increase our knowledge of the immune mechanisms involved in acute infection with non-cytopathic BVDV-1 strains, suggesting the existence of a clear tendency towards a type 1 immune response, thereby enhancing resistance against viral infections.

show abstract

Immunofluorescence Studies of Biotype-Specific Expression of Bovine Viral Diarrhoea Virus Epitopes in Infected Cells

Cited by 19 publications

References 18 publications

Uncleaved NS2-3 Is Required for Production of Infectious Bovine Viral Diarrhea Virus

Uncleaved NS2-3 Is Required for Production of Infectious Bovine Viral Diarrhea Virus

Formation of bovine viral diarrhea virus E1–E2 heterodimers is essential for virus entry and depends on charged residues in the transmembrane domains

Cell-Mediated Immune Response During Experimental Acute Infection with Bovine Viral Diarrhoea Virus: Evaluation of Blood Parameters

Contact Info

Product

Resources

About