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Characterization of monoclonal antibodies against foot-and-mouth disease virus serotype O and application in identification of antigenic variation in relation to vaccine strain selection.

Yang M, Xu W, Goolia M, Zhang Z - Virol. J. (2014)

Bottom Line: Isolates ECU/4/10 and HKN/2/11 demonstrated the highest antigenic variation compared to the others.The amino acid sequences of the outer capsid proteins for these nine isolates were analyzed and compared with the vaccine strain O1/Manisa.However, for vaccine matching VNT remains the preferred method but a combination of VNT, antigenic profiling with a panel of mAbs and genetic sequencing would probably be more ideal for full characterization of any new outbreak isolates as well as for selection of vaccine strains from FMDV antigen banks.

View Article: PubMed Central - HTML - PubMed

Affiliation: National Centre for Foreign Animal Disease, 1015 Arlington Street, Winnipeg R3E 3 M4, Manitoba, Canada. Ming.Yang@inspection.gc.ca.

ABSTRACT

Background: Foot-and-mouth disease (FMD) has severe implications for animal farming which leads to considerable financial losses because of its rapid spread, high morbidity and loss of productivity. For these reasons, the use of vaccine is often favoured to prevent and control FMD. Selection of the proper vaccine is extremely difficult because of the antigenic variation within FMDV serotypes. The aim of the current study was to produce a panel of mAbs and use it for the characterization of new isolates of FMDV serotype O.

Results: A panel of FMDV/O specific mAb was produced. The generated mAbs were then characterized using the peptide array and mAb resistant mutant selection. Seven out of the nine mAbs reacted with five known antigenic sites, thus the other two mAbs against non-neutralizing sites were identified. The mAbs were then evaluated by antigenic ELISA for the detection of forty-six FMDV serotype O isolates representing seven of ten known topotypes. Isolates ECU/4/10 and HKN/2/11 demonstrated the highest antigenic variation compared to the others. Furthermore, the panel of mAbs was used in vaccine matching by antigenic profiling ELISA with O1/Manisa as the reference strain. However, there was no correlation between vaccine matching by antigenic ELISA and the gold standard method, virus neutralisation test (VNT), for the forty-six FMDV/O isolates. Nine isolates had particularly poor correlation with the reference vaccine strain as revealed by the low r1 values in VNT. The amino acid sequences of the outer capsid proteins for these nine isolates were analyzed and compared with the vaccine strain O1/Manisa. The isolate ECU/4/10 displayed three unique amino acid substitutions around the antigenic sites 1, 3 and 4.

Conclusions: The panel of mAbs is useful to monitor the emergence of antigenically different strains and determination of relevant antigenic site differences. However, for vaccine matching VNT remains the preferred method but a combination of VNT, antigenic profiling with a panel of mAbs and genetic sequencing would probably be more ideal for full characterization of any new outbreak isolates as well as for selection of vaccine strains from FMDV antigen banks.

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Reactivity of mAbs (F12VP1O-2 and F21-64) with forty-one O/VP1 overlapping peptides in an indirect ELISA. Forty-one overlapping peptides and purified FMV/O were coated onto 96-well plate. The reactivities of the mAbs to the peptides and O1/BFS were detected with HRP anti-mouse IgG, followed by a substrate.
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Figure 1: Reactivity of mAbs (F12VP1O-2 and F21-64) with forty-one O/VP1 overlapping peptides in an indirect ELISA. Forty-one overlapping peptides and purified FMV/O were coated onto 96-well plate. The reactivities of the mAbs to the peptides and O1/BFS were detected with HRP anti-mouse IgG, followed by a substrate.

Mentions: In order to define the binding epitopes of the mAbs, the reactivity of the mAbs against recombinant VP1 and VP2 was examined using an indirect ELISA. Four mAbs reacted with the recombinant proteins (Table 1). The result showed that three mAbs (F12VP1O-2, F21-48 and F21-64) reacted with recombinant VP1 protein, while F11VP2O-2 reacted with recombinant VP2 protein (Table 1). This confirmed that the epitopes recognized by these four mAbs are linear, because the recombinant proteins were expressed in E. coli. The other mAbs failed to react with the recombinant proteins suggesting that the epitope recognized by these mAbs are conformational which is dependent on the integrity of viral particle structures. To further locate mAb binding epitopes, 41 peptides representing FMDV/O/VP1 and 42 peptides corresponding to O/VP2 were synthesized. Reactivity of the mAbs against peptides was examined using a peptide ELISA (Figure 1). The mAb F21-64 reacted with peptides 28-29 corresponding to the GH loop of VP1 at amino acids 136-151 (YSRNAVPNLRGDLQVL) which was recognized as the antigenic site1a. The mAb, F12VP1O-2 reacted with peptides 40-41 corresponding to the C-terminal residues of VP1 at amino acids 198-213 (EARHKQKIVAPVKQTL). This region was identified as the antigenic site1b, despite the fact that the mAb F12VP1O-2 did not demonstrate virus neutralization activity. The mAb F21-48 and F11VP2O-2 failed to react with any VP1/VP2 peptides, although it reacted with recombinant proteins.


Characterization of monoclonal antibodies against foot-and-mouth disease virus serotype O and application in identification of antigenic variation in relation to vaccine strain selection.

Yang M, Xu W, Goolia M, Zhang Z - Virol. J. (2014)

Reactivity of mAbs (F12VP1O-2 and F21-64) with forty-one O/VP1 overlapping peptides in an indirect ELISA. Forty-one overlapping peptides and purified FMV/O were coated onto 96-well plate. The reactivities of the mAbs to the peptides and O1/BFS were detected with HRP anti-mouse IgG, followed by a substrate.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4125342&req=5

Figure 1: Reactivity of mAbs (F12VP1O-2 and F21-64) with forty-one O/VP1 overlapping peptides in an indirect ELISA. Forty-one overlapping peptides and purified FMV/O were coated onto 96-well plate. The reactivities of the mAbs to the peptides and O1/BFS were detected with HRP anti-mouse IgG, followed by a substrate.
Mentions: In order to define the binding epitopes of the mAbs, the reactivity of the mAbs against recombinant VP1 and VP2 was examined using an indirect ELISA. Four mAbs reacted with the recombinant proteins (Table 1). The result showed that three mAbs (F12VP1O-2, F21-48 and F21-64) reacted with recombinant VP1 protein, while F11VP2O-2 reacted with recombinant VP2 protein (Table 1). This confirmed that the epitopes recognized by these four mAbs are linear, because the recombinant proteins were expressed in E. coli. The other mAbs failed to react with the recombinant proteins suggesting that the epitope recognized by these mAbs are conformational which is dependent on the integrity of viral particle structures. To further locate mAb binding epitopes, 41 peptides representing FMDV/O/VP1 and 42 peptides corresponding to O/VP2 were synthesized. Reactivity of the mAbs against peptides was examined using a peptide ELISA (Figure 1). The mAb F21-64 reacted with peptides 28-29 corresponding to the GH loop of VP1 at amino acids 136-151 (YSRNAVPNLRGDLQVL) which was recognized as the antigenic site1a. The mAb, F12VP1O-2 reacted with peptides 40-41 corresponding to the C-terminal residues of VP1 at amino acids 198-213 (EARHKQKIVAPVKQTL). This region was identified as the antigenic site1b, despite the fact that the mAb F12VP1O-2 did not demonstrate virus neutralization activity. The mAb F21-48 and F11VP2O-2 failed to react with any VP1/VP2 peptides, although it reacted with recombinant proteins.

Bottom Line: Isolates ECU/4/10 and HKN/2/11 demonstrated the highest antigenic variation compared to the others.The amino acid sequences of the outer capsid proteins for these nine isolates were analyzed and compared with the vaccine strain O1/Manisa.However, for vaccine matching VNT remains the preferred method but a combination of VNT, antigenic profiling with a panel of mAbs and genetic sequencing would probably be more ideal for full characterization of any new outbreak isolates as well as for selection of vaccine strains from FMDV antigen banks.

View Article: PubMed Central - HTML - PubMed

Affiliation: National Centre for Foreign Animal Disease, 1015 Arlington Street, Winnipeg R3E 3 M4, Manitoba, Canada. Ming.Yang@inspection.gc.ca.

ABSTRACT

Background: Foot-and-mouth disease (FMD) has severe implications for animal farming which leads to considerable financial losses because of its rapid spread, high morbidity and loss of productivity. For these reasons, the use of vaccine is often favoured to prevent and control FMD. Selection of the proper vaccine is extremely difficult because of the antigenic variation within FMDV serotypes. The aim of the current study was to produce a panel of mAbs and use it for the characterization of new isolates of FMDV serotype O.

Results: A panel of FMDV/O specific mAb was produced. The generated mAbs were then characterized using the peptide array and mAb resistant mutant selection. Seven out of the nine mAbs reacted with five known antigenic sites, thus the other two mAbs against non-neutralizing sites were identified. The mAbs were then evaluated by antigenic ELISA for the detection of forty-six FMDV serotype O isolates representing seven of ten known topotypes. Isolates ECU/4/10 and HKN/2/11 demonstrated the highest antigenic variation compared to the others. Furthermore, the panel of mAbs was used in vaccine matching by antigenic profiling ELISA with O1/Manisa as the reference strain. However, there was no correlation between vaccine matching by antigenic ELISA and the gold standard method, virus neutralisation test (VNT), for the forty-six FMDV/O isolates. Nine isolates had particularly poor correlation with the reference vaccine strain as revealed by the low r1 values in VNT. The amino acid sequences of the outer capsid proteins for these nine isolates were analyzed and compared with the vaccine strain O1/Manisa. The isolate ECU/4/10 displayed three unique amino acid substitutions around the antigenic sites 1, 3 and 4.

Conclusions: The panel of mAbs is useful to monitor the emergence of antigenically different strains and determination of relevant antigenic site differences. However, for vaccine matching VNT remains the preferred method but a combination of VNT, antigenic profiling with a panel of mAbs and genetic sequencing would probably be more ideal for full characterization of any new outbreak isolates as well as for selection of vaccine strains from FMDV antigen banks.

Show MeSH
Related in: MedlinePlus