Section 37
Chapter 36,718

Foot-and-mouth disease virus (FMDV) with a stable FLAG epitope in the VP1 G-H loop as a new tool for studying FMDV pathogenesis

Lawrence, P.; Pacheco, J.M.; Uddowla, S.; Hollister, J.; Kotecha, A.; Fry, E.; Rieder, E.

Virology 436(1): 150-161


ISSN/ISBN: 1096-0341
PMID: 23228860
DOI: 10.1016/j.virol.2012.11.001
Accession: 036717291

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Foot-and-mouth disease virus (FMDV) VP1 G-H loop contains the major antigenic site. By replacing the sequence upstream of the RGD motif with a FLAG epitope, a marker virus for pathogenesis studies was generated. In cell culture, the recombinant virus containing FLAG (A24-FLAG) exhibited similar plaque phenotypes and growth kinetics to parental virus. A24-FLAG was distinguished, neutralized, and immunoprecipitated by FLAG anti-sera. A24-FLAG infected cattle exhibited FMD and an antibody response similar to parental virus. FLAG epitope stability was confirmed both in vitro and in vivo. Interestingly, no anti-FLAG antibodies were detectable in cattle up to 21 days post-inoculation. A24-FLAG G-H loop modeling suggested FLAG was rendered a cryptic site, inaccessible to the host immune system. These studies demonstrate the FMDV VP1 G-H loop tolerance to substitutions without detriment to pathogenesis and antigenicity. Finally, A24-FLAG manifested virulence in cattle as parental virus, and could be distinguished and tracked by tag-specific anti-sera.

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