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Generation of replication-defective virus-based vaccines that confer full protection in sheep against virulent bluetongue virus challenge.

Authors
  • Matsuo, Eiko
  • Celma, Cristina C P
  • Boyce, Mark
  • Viarouge, Cyril
  • Sailleau, Corinne
  • Dubois, Eric
  • Bréard, Emmanuel
  • Thiéry, Richard
  • Zientara, Stéphan
  • Roy, Polly
Type
Published Article
Journal
Journal of Virology
Publisher
American Society for Microbiology
Publication Date
Oct 01, 2011
Volume
85
Issue
19
Pages
10213–10221
Identifiers
DOI: 10.1128/JVI.05412-11
PMID: 21795358
Source
Medline
License
Unknown

Abstract

The reverse genetics technology for bluetongue virus (BTV) has been used in combination with complementing cell lines to recover defective BTV-1 mutants. To generate a potential disabled infectious single cycle (DISC) vaccine strain, we used a reverse genetics system to rescue defective virus strains with large deletions in an essential BTV gene that encodes the VP6 protein (segment S9) of the internal core. Four VP6-deficient BTV-1 mutants were generated by using a complementing cell line that provided the VP6 protein in trans. Characterization of the growth properties of mutant viruses showed that each mutant has the necessary characteristics for a potential vaccine strain: (i) viral protein expression in noncomplementing mammalian cells, (ii) no infectious virus generated in noncomplementing cells, and (iii) efficient replication in the complementing VP6 cell line. Further, a defective BTV-8 strain was made by reassorting the two RNA segments that encode the two outer capsid proteins (VP2 and VP5) of a highly pathogenic BTV-8 with the remaining eight RNA segments of one of the BTV-1 DISC viruses. The protective capabilities of BTV-1 and BTV-8 DISC viruses were assessed in sheep by challenge with specific virulent strains using several assay systems. The data obtained from these studies demonstrated that the DISC viruses are highly protective and could offer a promising alternative to the currently available attenuated and killed virus vaccines and are also compliant as DIVA (differentiating infected from vaccinated animals) vaccines.

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