Identification of a novel viral protein in infectious bursal disease virus-infected cells

Infectious bursal disease virus (IBDV), the best characterized member of the Birnaviridae family, is a highly relevant avian pathogen causing both acute and persistent infections in different avian hosts. Here, we describe the establishment of clonal, long-term, productive persistent IBDV infections in DF-1 chicken embryonic fibroblasts. Although virus yields in persistently-infected cells are exceedingly lower than those detected in acutely infected cells, the replication fitness of viruses isolated from persistently-infected cells is higher than that of the parental virus. Persistently-infected DF-1 and IBDV-cured cell lines derived from them do not respond to type I interferon (IFN). High-throughput genome sequencing revealed that this defect is due to mutations affecting the IFNα/β receptor subunit 2 (IFNAR2) gene resulting in the expression of IFNAR2 polypeptides harbouring large C-terminal deletions that abolish the signalling capacity of IFNα/β receptor complex. Ectopic expression of a recombinant chicken IFNAR2 gene efficiently rescues IFNα responsiveness. IBDV-cured cell lines derived from persistently infected cells exhibit a drastically enhanced susceptibility to establishing new persistent IBDV infections. Additionally, experiments carried out with human HeLa cells lacking the IFNAR2 gene fully recapitulate results obtained with DF-1 cells, exhibiting a highly enhanced capacity to both survive the acute IBDV infection phase and to support the establishment of persistent IBDV infections. Results presented here show that the inactivation of the JAK-STAT signalling pathway significantly reduces the apoptotic response induced by the infection, hence facilitating the establishment and maintenance of IBDV persistent infections.IMPORTANCE Members of the Birnaviridae family, including infectious bursal disease virus (IBDV), exhibit a dual behaviour, causing acute infections that are often followed by the establishment of life-long persistent asymptomatic infections. Indeed, persistently infected specimens might act as efficient virus reservoirs, hence potentially contributing to virus dissemination. Despite the key importance of this biological trait, information about mechanisms triggering IBDV persistency is negligible. Our report evidences the capacity of IBDV, a highly relevant avian pathogen, to establishing long-term, productive, persistent infections in both avian and human cell lines. Data presented here provide novel and direct evidence about the crucial role of type I IFNs on the fate of IBDV-infected cells and their contribution to controlling the establishment of IBDV persistent infections. The use of cell lines unable to respond to type I IFNs opens a promising venue to unveiling additional factors contributing to IBDV persistency.

Download Full-text

Type I Interferon acts as a major barrier to the establishment of infectious bursal disease virus (IBDV) persistent infections

10.1101/2020.10.09.333161 ◽

2020 ◽

Author(s):

Laura Broto ◽

Nicolás Romero ◽

Fernando Méndez ◽

Elisabet Diaz-Beneitez ◽

Oscar Candelas-Rivera ◽

...

Keyword(s):

Disease Virus ◽

Cell Lines ◽

Infectious Bursal Disease Virus ◽

Infectious Bursal Disease ◽

Type I ◽

Persistently Infected ◽

Persistent Infections ◽

Avian Pathogen ◽

Bursal Disease ◽

Infected Cells

ABSTRACTInfectious bursal disease virus (IBDV), the best characterized member of the Birnaviridae family, is a highly relevant avian pathogen causing both acute and persistent infections in different avian hosts. Here, we describe the establishment of clonal, long-term, productive persistent IBDV infections in DF-1 chicken embryonic fibroblasts. Although virus yields in persistently-infected cells are exceedingly lower than those detected in acutely infected cells, the replication fitness of viruses isolated from persistently-infected cells is higher than that of the parental virus. Persistently-infected DF-1 and IBDV-cured cell lines derived from them do not respond to type I interferon (IFN). High-throughput genome sequencing revealed that this defect is due to mutations affecting the IFNα/β receptor subunit 2 (IFNAR2) gene resulting in the expression of IFNAR2 polypeptides harbouring large C-terminal deletions that abolish the signalling capacity of IFNα/β receptor complex. Ectopic expression of a recombinant chicken IFNAR2 gene efficiently rescues IFNα responsiveness. IBDV-cured cell lines derived from persistently infected cells exhibit a drastically enhanced proneness to establishing new persistent IBDV infections. Additionally, experiments carried out with human HeLa cells lacking the IFNAR2 gene fully recapitulate results obtained with DF-1 cells, exhibiting a highly enhanced capacity to both survive the acute IBDV infection phase and to support the establishment of persistent IBDV infections. Results presented here show that the inactivation of the JAK-STAT signalling pathway significantly reduces the apoptotic response induced by the infection, hence facilitating the establishment and maintenance of IBDV persistent infections.IMPORTANCEMembers of the Birnaviridae family, including infectious bursal disease virus (IBDV), exhibit a dual behaviour, causing acute infections that are often followed by the establishment of life-long persistent asymptomatic infections. Indeed, persistently infected specimens might act as efficient virus reservoirs, hence potentially contributing to virus dissemination. Despite the key importance of this biological trait, information about mechanisms triggering IBDV persistency is negligible. Our report evidences the capacity of IBDV, a highly relevant avian pathogen, to establishing long-term, productive, persistent infections in both avian and human cell lines. Data presented here provide novel and direct evidence about the crucial role of type I IFNs on the fate of IBDV-infected cells and their contribution to controlling the establishment of IBDV persistent infections. The use of cell lines unable to respond to type I IFNs opens a promising venue to unveiling additional factors contributing to IBDV persistency.

Download Full-text

Nuclear Factor NF45 Interacts with Viral Proteins of Infectious Bursal Disease Virus and Inhibits Viral Replication

Journal of Virology ◽

10.1128/jvi.02506-09 ◽

2010 ◽

Vol 84 (20) ◽

pp. 10592-10605 ◽

Cited By ~ 30

Author(s):

Ruth L. O. Stricker ◽

Sven-Erik Behrens ◽

Egbert Mundt

Keyword(s):

Viral Replication ◽

Disease Virus ◽

Infectious Bursal Disease Virus ◽

Molecular Mechanisms ◽

Viral Proteins ◽

Infectious Bursal Disease ◽

Nuclear Factor ◽

Replication Process ◽

Bursal Disease ◽

Infected Cells

ABSTRACT Two of the central issues in developing new strategies to interfere with viral infections concern the identification of cellular proteins involved in viral replication and/or antiviral measures and the dissection of the underlying molecular mechanisms. To gain initial insight into the role of host proteins in the life cycle of infectious bursal disease virus (IBDV), a double-stranded RNA virus, we examined the cellular nuclear factor 45 (NF45). NF45 was previously indicated to be involved in the replication process of other types of RNA viruses. Interestingly, by performing immunofluorescence studies, we found that in IBDV-infected cells the mainly nuclear NF45 accumulated at the sites of viral replication in the cytoplasm. NF45 was shown to specifically colocalize with the viral RNA-dependent RNA polymerase VP1, the capsid protein VP2, and the ribonucleoprotein VP3. Immunoprecipitation experiments indicated protein-protein associations between NF45 and VP1, VP2, and VP3. Expression of the individual VP3 or the combination of expression of VP1 and VP3 did not result in a cytoplasmic accumulation of NF45, which, among other data, showed that recruitment of the cellular protein in infected cells functionally correlates with the viral replication process. Since small interfering RNA(siRNA)-mediated downregulation of NF45 resulted in an approximately 5-fold increase of virus yield, our study suggests that NF45, by association with viral proteins, is part of a yet-uncharacterized cellular defense mechanism against IBDV infections.

Download Full-text

Interactions in vivo between the proteins of infectious bursal disease virus: capsid protein VP3 interacts with the RNA-dependent RNA polymerase, VP1

Microbiology ◽

10.1099/0022-1317-81-1-209 ◽

2000 ◽

Vol 81 (1) ◽

pp. 209-218 ◽

Cited By ~ 45

Author(s):

Mirriam G. J. Tacken ◽

Peter J. M. Rottier ◽

Arno L. J. Gielkens ◽

Ben P. H. Peeters

Keyword(s):

Disease Virus ◽

Protein Interactions ◽

Infectious Bursal Disease Virus ◽

Viral Proteins ◽

Infectious Bursal Disease ◽

Transactivation Domain ◽

Protein Protein Interactions ◽

Bursal Disease ◽

Infected Cells

Little is known about the intermolecular interactions between the viral proteins of infectious bursal disease virus (IBDV). By using the yeast two-hybrid system, which allows the detection of protein–protein interactions in vivo, all possible interactions were tested by fusing the viral proteins to the LexA DNA-binding domain and the B42 transactivation domain. A heterologous interaction between VP1 and VP3, and homologous interactions of pVP2, VP3, VP5 and possibly VP1, were found by co-expression of the fusion proteins in Saccharomyces cerevisiae. The presence of the VP1–VP3 complex in IBDV-infected cells was confirmed by co-immunoprecipitation studies. Kinetic analyses showed that the complex of VP1 and VP3 is formed in the cytoplasm and eventually is released into the cell-culture medium, indicating that VP1–VP3 complexes are present in mature virions. In IBDV-infected cells, VP1 was present in two forms of 90 and 95 kDa. Whereas VP3 initially interacted with both the 90 and 95 kDa proteins, later it interacted exclusively with the 95 kDa protein both in infected cells and in the culture supernatant. These results suggest that the VP1–VP3 complex is involved in replication and packaging of the IBDV genome.

Download Full-text

The association of ribosomal protein L18 (RPL18) with infectious bursal disease virus viral protein VP3 enhances viral replication

Virus Research ◽

10.1016/j.virusres.2017.12.009 ◽

2018 ◽

Vol 245 ◽

pp. 69-79 ◽

Cited By ~ 4

Author(s):

Bin Wang ◽

Xueyan Duan ◽

Mengjiao Fu ◽

Yanan Liu ◽

Yongqiang Wang ◽

...

Keyword(s):

Viral Replication ◽

Ribosomal Protein ◽

Disease Virus ◽

Viral Protein ◽

Infectious Bursal Disease Virus ◽

Infectious Bursal Disease ◽

Bursal Disease

Download Full-text

The VP5 protein of infectious bursal disease virus promotes virion release from infected cells and is not involved in cell death

Archives of Virology ◽

10.1007/s00705-009-0524-4 ◽

2009 ◽

Vol 154 (12) ◽

pp. 1873-1882 ◽

Cited By ~ 27

Author(s):

Yongping Wu ◽

Lianlian Hong ◽

Juxiu Ye ◽

Zhenyu Huang ◽

Jiyong Zhou

Keyword(s):

Cell Death ◽

Disease Virus ◽

Infectious Bursal Disease Virus ◽

Infectious Bursal Disease ◽

Virion Release ◽

Bursal Disease ◽

Infected Cells

Download Full-text

RNA-Dependent RNA polymerase activity of the infectious bursal disease virus viral protein 1

10.5353/th_b3040819 ◽

2004 ◽

Author(s):

Hok-tsun Ma

Keyword(s):

Rna Polymerase ◽

Disease Virus ◽

Viral Protein ◽

Infectious Bursal Disease Virus ◽

Polymerase Activity ◽

Infectious Bursal Disease ◽

Rna Dependent Rna Polymerase ◽

Bursal Disease ◽

Rna Polymerase Activity ◽

Viral Protein 1

Download Full-text

Viral Protein 1 Sequence Analysis of Three Infectious Bursal Disease Virus Strains: A Very Virulent Virus, Its Attenuated Form, and an Attenuated Vaccine

Avian Diseases ◽

10.2307/1592762 ◽

1999 ◽

Vol 43 (1) ◽

pp. 55 ◽

Cited By ~ 9

Author(s):

Hila Yehuda ◽

Jacob Pitcovski ◽

Amnon Michael ◽

Bezalel Gutter ◽

Martin Goldway

Keyword(s):

Sequence Analysis ◽

Disease Virus ◽

Viral Protein ◽

Infectious Bursal Disease Virus ◽

Infectious Bursal Disease ◽

Attenuated Vaccine ◽

Bursal Disease ◽

Virulent Virus ◽

Viral Protein 1 ◽

Virus Strains

Download Full-text

Effect of Soybean Isoflavones on Growth Performance, Immune Function, and Viral Protein 5 mRNA Expression in Broiler Chickens Challenged with Infectious Bursal Disease Virus

Animals ◽

10.3390/ani9050247 ◽

2019 ◽

Vol 9 (5) ◽

pp. 247

Author(s):

Mahmoud Mostafa Azzam ◽

Shou-qun JIANG ◽

Jia-li CHEN ◽

Xia-jing LIN ◽

Zhong-yong GOU ◽

...

Keyword(s):

Antioxidant Activity ◽

Mrna Expression ◽

Disease Virus ◽

Viral Protein ◽

Infectious Bursal Disease Virus ◽

Broiler Chickens ◽

Infected Group ◽

Infectious Bursal Disease ◽

Infected Control ◽

Bursal Disease

A total of 200 one-day-old male broilers were assigned to five groups, and each group consisted of four replicates with 10 birds per replicate. Chicks were fed the basal diet with 0 (non-infected control), 0 (infected control), 10, 20, and 40 mg/kg soybean isoflavones (SI) for 42 days. At 21 days of age, chickens were inoculated with an infectious bursal dose (causing 50% morbidity) of the infectious bursal disease virus (IBDV) BC 6/85 strain by the eye-drop and nasal route (except for the non-infected group). Average daily gain (ADG) and average daily feed intake (ADFI) decreased (p < 0.05) in broilers infected with infectious bursal disease virus (IBDV) from 22 to 42 days. However, infected broilers fed 10 and 20 mg SI/kg had the maximum (p <0.05) ADG and ADFI from 1 to 42 days. Body weight (BW) increased (p < 0.05) in infected broilers fed the 10 and 20 mg SI /kg diet. The bursa weight at 7 days post-infection (dpi) was increased (p < 0.05) by the supplemental 10 mg SI/kg diet. Infected broilers showed the highest (p < 0.05) bursa lesions, with an average score of 4.0 ± 0.0, while the severity of bursa lesions was decreased (p < 0.05) at 3 dpi and 7 dpi by the supplemental 20 mg SI/kg diet. Supplemental SI at 20 mg/kg decreased (p < 0.05) the viral protein 5 (VP5) mRNA expression at 3 dpi and 7 dpi. The level of interferon gamma (IFNγ) was elevated (p < 0.05) in the infected group at 3 dpi and 7 dpi as compared with the control group, while its level was decreased (p < 0.05) by supplemental 10 mg/kg SI at 3 dpi. The level of nuclear factor κB in the bursal tissue showed the lowest value (p < 0.05) with supplemental 10 and 20 mg SI/kg diet at 7 dpi. Supplemental 10, 20, 40 mg/kg SI improved (p < 0.05) the serum total antioxidant activity (T-AOC) in infected broilers at 3 dpi. In addition, the serum level of malondialdehyde (MDA) decreased (p < 0.05) in the group fed 20 mg/kg SI at 7 dpi. In conclusion, supplemental 10~20 mg/kg SI may have a positive effect on broiler chickens infected with IBDV, probably because SI decrease the severity of bursa lesions and viral protein 5 mRNA expression, and have strong antioxidant activity.

Download Full-text