Capsid proteins from field strains of foot-and-mouth disease virus confer a pathogenic phenotype in cattle on an attenuated, cell-culture-adapted virus

Bøtner, Anette; Kakker, N. K.; Barbezange, Cyril; Berryman, Stephen; Jackson, Terry; Belsham, Graham J.

doi:10.1099/vir.0.029710-0

Cited by 34 publications

(43 citation statements)

References 34 publications

(52 reference statements)

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“…Previously, we described a recombinant virus (O1K-A/BTY1) recovered from an infectious copy plasmid (pO1K-A) by transfection of in vitro-transcribed vRNA into BHK cells and one passage through pBTY cells (37). As expected for a virus with a capsid derived from a field isolate, O1K-A/BTY1 was infectious for pBTY and BHK cells but was not infectious for CHO cells (Table 2), as they lack all of the known integrin receptors of FMDV.…”

Section: Resultsmentioning

confidence: 99%

“…Construction of infectious copy plasmids O1K-A and O1K-OUK has been described in detail previously (37). These plasmids are based on the infectious copy plasmid pT7S3, which carries a cDNA copy of the full-length viral RNA for FMDV O1K/B64.…”

Section: Methodsmentioning

confidence: 99%

“…Virus (O1KA-A/BTY1) recovered from this plasmid caused normal signs of FMD in cattle and pigs (37,38). Here, we studied cell culture adaption of O1K-A/BTY1 and identified a single amino acid substitution (glutamine [Q] to lysine [K]) at VP1-110 that allows for integrin-and HS-independent infection of CHO cells.…”

mentioning

confidence: 99%

“…We previously described an FMDV infectious copy plasmid (pO1K-A) that carries genes for capsid proteins VP1, VP2, and VP3 of the A/Turkey/2/2006 field strain combined with VP4 and the nonstructural proteins of a cell culture-adapted type O virus, O 1 Kaufbeuren B64 (O1KB64) (37). Virus (O1KA-A/BTY1) recovered from this plasmid caused normal signs of FMD in cattle and pigs (37,38).…”

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confidence: 99%

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Positively Charged Residues at the Five-Fold Symmetry Axis of Cell Culture-Adapted Foot-and-Mouth Disease Virus Permit Novel Receptor Interactions

Berryman

Clark

Kakker

et al. 2013

J Virol

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Field isolates of foot-and-mouth disease virus (FMDVF oot-and-mouth disease (FMD) is endemic in many regions of the world and is one of the most widespread, epizootic transboundary animal diseases, affecting many species of wildlife and livestock, such as cattle, sheep, goats, and pigs. The significant economic losses that result from FMD are due to the high morbidity of infected animals and stringent trade restrictions imposed on affected countries (1). Vaccination plays a major role in controlling FMD, either to lessen the effects of an outbreak in FMDfree countries or for control and eradication in regions where it is endemic. The etiological agent of FMD, foot-and-mouth disease virus (FMDV), exists as seven distinct serotypes (O, A, C, Asia-1, and the Southern African Territories [SAT] serotypes SAT-1, SAT-2, and SAT-3). Within each serotype, a large number of antigenic variants exist (2). Intraserotype diversity is driven by a high mutation rate during replication that is caused by an error-prone viral RNA-dependent RNA polymerase (3) and thus complicates efforts to control disease by vaccination due to incomplete protection between some antigenic variants (4). Hence, the most effective vaccines closely match the outbreak virus, which can necessitate the development of new vaccine strains. The current vaccines are inactivated virus preparations grown in large-scale cell culture. Therefore, the production of a new vaccine is critically dependent upon adaptation of viruses from the field for growth in cell culture, which can prove problematical for some viruses.Foot-and-mouth disease virus is the type species of the Aphthovirus genus of the Picornaviridae, a family of nonenveloped, single-stranded positive-sense RNA viruses. The viral capsid is formed by 60 copies each of four structural proteins (VP1 to VP4) arranged in icosahedral symmetry. The outer capsid surfaces are formed by VP1, which surrounds the five-fold symmetry axis, and VP2 and VP3, which alternate around the three-fold axis (5). VP4 is myristoylated and located inside the capsid and is thought to play an essential role in the final stage of assembly and in endosomal membrane penetration by the viral RNA (6, 7). In vivo, FMDV has a strong tropism for epithelial cells, which is in part due to the epithelial cell-restricted expression of integrin ␣v␤6, which is the principal receptor used by field viruses to initiate infection (8-12). Integrin binding is mediated by a highly conserved arginine-glycine-aspartic acid (RGD) motif located at the apex of a structurally disordered loop (the GH loop of VP1). The integrin specificity of FMDV has been the subject of several studies, and three other RGD-dependent integrins (␣v␤1, ␣v␤3, and ␣v␤8) have also been reported to be receptors for field strains of the virus (13-15); however, the role of these integrins in pathogenesis is unclear, and we have found that ␣v␤3 is a poor receptor for FMDV in vitro (16). Furthermore, despite recognizing their ligands via the RGD motif, two other RGD-dependent integrins ...

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Positively Charged Residues at the Five-Fold Symmetry Axis of Cell Culture-Adapted Foot-and-Mouth Disease Virus Permit Novel Receptor Interactions

Berryman

Clark

Kakker

et al. 2013

J Virol

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“…Plasmids containing the FMDV O1Kaufbeuren (O1K) cDNA have been described previously (39,40). The cDNA corresponding to the O1 Manisa VP2-2A coding region was amplified from pGEM-3Z-O-P1-2A (31) and pGEM-3Z-O-P1-2A-mIRES-3C VP1K210E (described above) using primers FMDVO_NheIVP4VP2_Fw and FMDVO_ApaI2A2B_Re ( Table 1).…”

Section: Methodsmentioning

confidence: 99%

Processing of the VP1/2A Junction Is Not Necessary for Production of Foot-and-Mouth Disease Virus Empty Capsids and Infectious Viruses: Characterization of “Self-Tagged” Particles

Gullberg

Polacek

Bøtner

et al. 2013

J Virol

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The foot-and-mouth disease virus (FMDV) capsid protein precursor, P1-2A, is cleaved by 3C pro to generate VP0, VP3, VP1, and the peptide 2A. The capsid proteins self-assemble into empty capsid particles or viruses which do not contain 2A. In a cell culture-adapted strain of FMDV (O1 Manisa [Lindholm]), three different amino acid substitutions (E83K, S134C, and K210E) were identified within the VP1 region of the P1-2A precursor compared to the field strain (wild type [wt]). Expression of the O1 Manisa P1-2A (wt or with the S134C substitution in VP1) plus 3C pro , using a transient expression system, resulted in efficient capsid protein production and self-assembly of empty capsid particles. Removal of the 2A peptide from the capsid protein precursor had no effect on capsid protein processing or particle assembly. However, modification of E83K alone abrogated particle assembly with no apparent effect on protein processing. Interestingly, the K210E substitution, close to the VP1/2A junction, completely blocked processing by 3C pro at this cleavage site, but efficient assembly of "self-tagged" empty capsid particles, containing the uncleaved VP1-2A, was observed. These self-tagged particles behaved like the unmodified empty capsids in antigen enzyme-linked immunosorbent assays and integrin receptor binding assays. Furthermore, mutant viruses with uncleaved VP1-2A could be rescued in cells from full-length FMDV RNA transcripts encoding the K210E substitution in VP1. Thus, cleavage of the VP1/2A junction is not essential for virus viability. The production of such engineered self-tagged empty capsid particles may facilitate their purification for use as diagnostic reagents and vaccines.

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Megaprimer-mediated capsid swapping for the construction of custom-engineered chimeric foot-and-mouth disease virus

et al. 2015

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Foot-and-mouth disease (FMD) is a highly contagious, economically important disease of transboundary importance. Regular vaccination with chemically inactivated FMD vaccine is the major means of controlling the disease in endemic countries like India. However, the selection of appropriate candidate vaccine strain and its adaptation in cell culture to yield high titer of virus is a cumbersome process. An attractive approach to circumvent this tedious process is to replace the capsid coding sequence of an infectious full-genome length cDNA clone of a good vaccine strain with those of appropriate field strain, to produce custom-made chimeric FMD virus (FMDV). Nevertheless, the construction of chimeric virus can be difficult if the necessary endonuclease restriction sites are unavailable or unsuitable for swapping of the capsid sequence. Here we described an efficient method based on megaprimer-mediated capsid swapping for the construction of chimeric FMDV cDNA clones. Using FMDV vaccine strain A IND 40/2000 infectious clone (pA(40/2000)) as a donor plasmid, we exchanged the capsid sequence of pA(40/2000) with that of the viruses belonging to serotypes O (n = 5), A (n = 2), and Asia 1 (n = 2), and subsequently generated infectious FMDV from their respective chimeric cDNA clones. The chimeric viruses exhibited comparable infection kinetics, plaque phenotypes, antigenic profiles, and virion stability to the parental viruses. The results from this study suggest that megaprimer-based reverse genetics technology is useful for engineering chimeric vaccine strains for use in the control and prevention of FMD in endemic countries.

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Capsid proteins from field strains of foot-and-mouth disease virus confer a pathogenic phenotype in cattle on an attenuated, cell-culture-adapted virus

Cited by 34 publications

References 34 publications

Positively Charged Residues at the Five-Fold Symmetry Axis of Cell Culture-Adapted Foot-and-Mouth Disease Virus Permit Novel Receptor Interactions

Positively Charged Residues at the Five-Fold Symmetry Axis of Cell Culture-Adapted Foot-and-Mouth Disease Virus Permit Novel Receptor Interactions

Processing of the VP1/2A Junction Is Not Necessary for Production of Foot-and-Mouth Disease Virus Empty Capsids and Infectious Viruses: Characterization of “Self-Tagged” Particles

Megaprimer-mediated capsid swapping for the construction of custom-engineered chimeric foot-and-mouth disease virus

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