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Analysis of foot-and-mouth disease virus type O1 Brugge neutralization epitopes using monoclonal antibodies
Abstract:
Monoclonal antibodies (MAbs) were elicited with inactivated, purified foot-and-mouth disease virus (FMDV) type O1 strain Brugge (140S) and with 12S protein subunits. Each MAb was tested for its capacity to bind to FMDV O1 Brugge 140S virions, 12S subunits and purified VP1 by radioimmunoassay (RIA) and to neutralize viral infectivity in mouse protection assays. Those MAbs which reacted only with 12S subunits in RIA did not neutralize infectious virus. One MAb, 12FE9.2.1, reacted with 140S, 12S and purified VP1 of FMDV O1 Brugge and neutralized infectious virus. Reactions with different biosynthetic FMDV type O1 Campos VP1 polypeptides localized the binding site of 12FE9.2.1 between amino acid residues 135 and 172. Eight MAbs reacted with both 140S virions and 12S subunits and neutralized infectious virus. Monoclonal antibodies that reacted only with 12S protein subunits or with 140S, 12S and VP1 did not compete in RIA with 140S/12S-reactive neutralizing MAbs for 12S binding sites. The ability of 140S/12S-reactive MAbs to compete for the 12S binding site defined by MAb 10GA4.2.2 was directly related to their capacity to neutralize infectious virus, suggesting that these MAbs were all affecting a single neutralization site. In mouse protection and plaque reduction neutralization assays, 10GA4.2.2 and 12FE9.2.1 were equally effective in neutralizing the homologous FMDV type O1 Brugge, and in cross-neutralization assays both exhibited high titres against four additional strains of type O1 FMDV.
Insights
Monoclonal antibodies (MAbs) targeting foot-and-mouth disease virus (FMDV) were developed. Certain MAbs neutralizing FMDV were identified, binding to specific viral components and offering protection in mouse models.
Area of Science:
- Immunology
- Virology
- Molecular Biology
Background:
- Foot-and-mouth disease virus (FMDV) poses a significant threat to livestock globally.
- Developing effective neutralizing antibodies is crucial for FMDV control strategies.
- Understanding antibody-virus interactions at the molecular level aids in vaccine and therapeutic development.
Purpose of the Study:
- To generate and characterize monoclonal antibodies (MAbs) against FMDV type O1 strain Brugge.
- To identify MAbs capable of neutralizing FMDV infectivity.
- To map the binding sites of neutralizing MAbs on FMDV structural components.
Main Methods:
- Production of MAbs using inactivated FMDV (140S) and 12S subunits.
- Radioimmunoassay (RIA) to assess MAb binding to FMDV virions, 12S subunits, and VP1.
- Mouse protection assays and plaque reduction neutralization assays to evaluate viral neutralization.
- Epitope mapping using biosynthetic VP1 polypeptides.
Main Results:
- MAbs reacting solely with 12S subunits did not neutralize infectious virus.
- One MAb, 12FE9.2.1, bound to 140S, 12S, and VP1, and neutralized FMDV.
- The binding site of 12FE9.2.1 was localized to VP1 residues 135-172.
- Eight MAbs reacted with both 140S and 12S subunits and neutralized FMDV.
- Neutralizing MAbs targeting the 12S binding site showed direct correlation with neutralization capacity, indicating a single neutralization site.
Conclusions:
- Specific MAbs targeting FMDV structural proteins, particularly VP1, can neutralize viral infectivity.
- The identified neutralization site on FMDV is critical for viral infectivity.
- These neutralizing MAbs demonstrate potential for FMDV diagnostics and therapeutics, with cross-neutralization activity against multiple type O1 strains.