Simultaneous alteration of residues 279 and 284 of the VP2 major capsid protein of a very virulent Infectious Bursal Disease Virus (vvIBDV) strain did not lead to attenuation in chickens

Virology Journal

Helmi Mardassi²  Abdeljelil Ghram³  Samir Boubaker¹  Khaled Miled³  Selma Kassar¹  Neila Khabouchi²  Nawel Ben Abdeljelil² 

[1] Laboratoire d’Anatomie Pathologique Humaine et Expérimentale, Université de Tunis El Manar, Institut Pasteur de Tunis, 13, Place Pasteur BP 74, Tunis Belvedere, 1002, Tunis, Tunisie;LR11IPT01 Laboratory of Molecular Microbiology, Vaccinology and Biotechnology Development, Université de Tunis El Manar, Institut Pasteur de Tunis, 13, Place Pasteur BP 74, Tunis Belvedere, 1002, Tunis, Tunisie;LR11IPT03 Laboratory of Veterinary Microbiology, Institut Pasteur de Tunis, 13, Place Pasteur BP 74, Tunis Belvedere, 1002, Tunis, Tunisie

关键词: Attenuation;    Virulence;    Tropism;    Mutagenesis;    In vivo reverse genetics;    Infectious clone;    vvIBDV;   

Others  :  1131089
DOI  :  10.1186/s12985-014-0199-7

 received in 2014-06-02, accepted in 2014-11-07,  发布年份 2014

【 摘 要 】

Background

Cell culture adaptation of very virulent infectious bursal disease virus (vvIBDV) was shown to be mainly associated with the VP2 capsid protein residues 253, 279, and 284. The single mutation A284T proved critical for cell culture tropism, but did not confer efficient virus replication, which at least required one additional mutation, Q253H or D279N. While the double mutation Q253H/A284T was unambiguously shown to confer both efficient replication in cell culture and attenuation in chickens, conflicting results have been reported regarding the replication efficiency of vvIBDV mutants bearing the D279N/A284T double mutation, and no data are hitherto available on their virulence in chickens.

Findings

Here we used an in vivo reverse genetics system to assess the impact of the D279N/A284T double mutation on the replication and attenuation of a chimeric IBDV virus, whose polyprotein derived from a non-culturable vvIBDV clinical isolate. We found that the D279N/A284T double mutation did indeed confer efficient replication in chicken embryo fibroblast (CEF) cell culture, but the mutant virus remained highly pathogenic to chickens.

Conclusions

The double mutation D279N/A284T of the VP2 major capsid protein of vvIBDV is sufficient to confer cell culture tropism and replication efficiency, but does not necessarily lead to virus attenuation.

【 授权许可】

   
2014 Ben Abdeljelil et al.; licensee BioMed Central Ltd.

【 图 表 】

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【 参考文献 】

• [1]Becht H: Infectious bursal disease virus. Curr Top Microbiol Immunol 1980, 90:107-121.
• [2]Müller H, Islam MR, Raue R: Research on infectious bursal disease–the past, the present and the future. Vet Microbiol 2003, 97:153-165.
• [3]Hirai K, Shimakura S, Kawamoto E, Taguchi F, Kim ST, Chang CN, Iritani Y: The immunodepressive effect of infectious bursal disease virus in chickens. Avian Dis 1974, 18:50-57.
• [4]Meulemans G, Halen P, Vindevogel H, Gouffaux M: The immunodepressive effect of infectious bursal agent on vaccination against Newcastle disease. Res Vet Sci 1977, 22:222-224.
• [5]Vasconcelos AC, Lam KM: Apoptosis in chicken embryos induced by the infectious bursal disease virus. J Comp Pathol 1995, 112:327-338.
• [6]Muller H, Mundt E, Eterradossi N, Islam MR: Current status of vaccines against infectious bursal disease. Avian Pathol 2012, 41:133-139.
• [7]Dobos P, Hill BJ, Hallett R, Kells DT, Becht H, Teninges D: Biophysical and biochemical characterization of five animal viruses with bisegmented double-stranded RNA genomes. J Virol 1979, 32:593-605.
• [8]Muller H, Scholtissek C, Becht H: The genome of infectious bursal disease virus consists of two segments of double-stranded RNA. J Virol 1979, 31:584-589.
• [9]Spies U, Muller H, Becht H: Properties of RNA polymerase activity associated with infectious bursal disease virus and characterization of its reaction products. Virus Res 1987, 8:127-140.
• [10]von Einem UI, Gorbalenya AE, Schirrmeier H, Behrens SE, Letzel T, Mundt E: VP1 of infectious bursal disease virus is an RNA-dependent RNA polymerase. J Gen Virol 2004, 85:2221-2229.
• [11]Garriga D, Navarro A, Querol-Audi J, Abaitua F, Rodriguez JF, Verdaguer N: Activation mechanism of a noncanonical RNA-dependent RNA polymerase. Proc Natl Acad Sci U S A 2007, 104:20540-20545.
• [12]Hudson PJ, McKern NM, Power BE, Azad AA: Genomic structure of the large RNA segment of infectious bursal disease virus. Nucleic Acids Res 1986, 14:5001-5012.
• [13]Mundt E, Beyer J, Muller H: Identification of a novel viral protein in infectious bursal disease virus-infected cells. J Gen Virol 1995, 76:437-443.
• [14]Muller H, Becht H: Biosynthesis of virus-specific proteins in cells infected with infectious bursal disease virus and their significance as structural elements for infectious virus and incomplete particles. J Virol 1982, 44:384-392.
• [15]Birghan C, Mundt E, Gorbalenya AE: A non-canonical lon proteinase lacking the ATPase domain employs the ser-Lys catalytic dyad to exercise broad control over the life cycle of a double-stranded RNA virus. EMBO J 2000, 19:114-123.
• [16]Casañas A, Navarro A, Ferrer-Orta C, González D, Rodríguez JF, Verdaguer N: Structural insights into the multifunctional protein VP3 of birnaviruses. Structure 2008, 16:29-37.
• [17]Jackwood DJ, Saif YM, Hughes JH: Characteristics and serologic studies of two serotypes of infectious bursal disease virus in turkeys. Avian Dis 1982, 26:871-882.
• [18]McNulty MS, Saif YM: Antigenic relationship of non-serotype 1 turkey infectious bursal disease viruses from the United States and United Kingdom. Avian Dis 1988, 32:374-375.
• [19]Ismail NM, Saif YM, Moorhead PD: Lack of pathogenicity of five serotype 2 infectious bursal disease viruses in chickens. Avian Dis 1988, 32:757-759.
• [20]Berg TP, Meulemans G: Acute infectious bursal disease in poultry: protection afforded by maternally derived antibodies and interference with live vaccination. Avian Pathol 1991, 20:409-421.
• [21]Chettle NJ, Wyeth PJ: Failure of maternally derived infectious bursal disease antibodies to serotypes 1 and 2 to protect against heterologous virus. Br Vet J 1989, 145:165-169.
• [22]Di Fabio J, Rossini LI, Eterradossi N, Toquin MD, Gardin Y: European-like pathogenic infectious bursal disease viruses in Brazil. Vet Rec 1999, 145:203-204.
• [23]Eterradossi N, Picault JP, Drouin P, Guittet M, L’Hospitalier R, Bennejean G: Pathogenicity and preliminary antigenic characterization of six infectious bursal disease virus strains isolated in France from acute outbreaks. Zentralbl Veterinarmed B 1992, 39:683-691.
• [24]Eterradossi N, Arnauld C, Tekaia F, Toquin D, Le Coq H, Rivallan G, Guitet M, Domenech J, Van den Berg TP, Skinner MA: Antigenic and genetic relationships between European very virulent infectious bursal disease viruses and an early West African isolate. Avian Pathol 1999, 28:36-46.
• [25]Nakamura T, Otaki Y, Nunoya T: Immunosuppressive effect of a highly virulent infectious bursal disease virus isolated in Japan. Avian Dis 1992, 36:891-896.
• [26]Brandt M, Yao K, Liu M, Heckert RA, Vakharia VN: Molecular determinants of virulence, cell tropism, and pathogenic phenotype of infectious bursal disease virus. J Virol 2001, 75:11974-11982.
• [27]Mundt E: Tissue culture infectivity of different strains of infectious bursal disease virus is determined by distinct amino acids in VP2. J Gen Virol 1999, 80:2067-2076.
• [28]van Loon AA, de Haas N, Zeyda I, Mundt E: Alteration of amino acids in VP2 of very virulent infectious bursal disease virus results in tissue culture adaptation and attenuation in chickens. J Gen Virol 2002, 83:121-129.
• [29]Lim BL, Cao Y, Yu T, Mo CW: Adaptation of very virulent infectious bursal disease virus to chicken embryonic fibroblasts by site-directed mutagenesis of residues 279 and 284 of viral coat protein VP2. J Virol 1999, 73:2854-2862.
• [30]Qi X, Gao H, Gao Y, Qin L, Wang Y, Gao L, Wang X: Naturally occurring mutations at residues 253 and 284 in VP2 contribute to the cell tropism and virulence of very virulent infectious bursal disease virus. Antiviral Res 2009, 84:225-233.
• [31]Qi X, Zhang L, Chen Y, Gao L, Wu G, Qin L, Wang Y, Ren X, Gao Y, Gao H, Wang X: Mutations of residues 249 and 256 in VP2 are involved in the replication and virulence of infectious Bursal disease virus. PLoS One 2013, 8:e70982.
• [32]Ben Abdeljelil N, Khabouchi N, Mardassi H: Efficient rescue of infectious bursal disease virus using a simplified RNA polymerase II-based reverse genetics strategy. Arch Virol 2008, 153:1131-1137.
• [33]Ben Abdeljelil N, Delmas B, Mardassi H: Replication and packaging of an infectious bursal disease virus segment A-derived minigenome. Virus Res 2008, 136:146-151.
• [34]Mundt E, Vakharia VN: Synthetic transcripts of double-stranded Birnavirus genome are infectious. Proc Natl Acad Sci U S A 1996, 93:11131-11136.
• [35]Mardassi H, Khabouchi N, Ghram A, Namouchi A, Karboul A: A very virulent genotype of infectious bursal disease virus predominantly associated with recurrent infectious bursal disease outbreaks in Tunisian vaccinated flocks. Avian Dis 2004, 48:829-840.
• [36]Ho SN, Hunt HD, Horton RM, Pullen JK, Pease LR: Site-directed mutagenesis by overlap extension using the polymerase chain reaction. Gene 1989, 77:51-59.
• [37]Nouen CL, Toquin D, Muller H, Raue R, Kean KM, Langlois P, Cherbonnel M, Eterradossi N: Different domains of the RNA polymerase of infectious bursal disease virus contribute to virulence. PLoS One 2012, 7:e28064.
• [38]Ismail NM, Saif YM: Immunogenicity of infectious bursal disease viruses in chickens. Avian Dis 1991, 35:460-469.
• [39]Raue R, Islam MR, Islam MN, Islam KM, Badhy SC, Das PM, Muller H: Reversion of molecularly engineered, partially attenuated, very virulent infectious bursal disease virus during infection of commercial chickens. Avian Pathol 2004, 33:181-189.
• [40]Yamaguchi T, Ogawa M, Inoshima Y, Miyoshi M, Fukushi H, Hirai K: Identification of sequence changes responsible for the attenuation of highly virulent infectious bursal disease virus. Virology 1996, 223:219-223.
• [41]Escaffre O, Le Nouen C, Amelot M, Ambroggio X, Ogden KM, Guionie O, Toquin D, Muller H, Islam MR, Eterradossi N: Both genome segments contribute to the pathogenicity of very virulent infectious bursal disease virus. J Virol 2013, 87:2767-2780.
• [42]Yu F, Ren X, Wang Y, Qi X, Song J, Gao Y, Qin L, Gao H, Wang X: A single amino acid V4I substitution in VP1 attenuates virulence of very virulent infectious bursal disease virus (vvIBDV) in SPF chickens and increases replication in CEF cells. Virology 2013, 440:204-209.
• [43]Gao L, Li K, Qi X, Gao H, Gao Y, Qin L, Wang Y, Shen N, Kong X, Wang X: Triplet amino acids located at positions 145/146/147 of the RNA polymerase of very virulent infectious bursal disease virus contribute to viral virulence. J Gen Virol 2014, 95(Pt 4):888-897.