J Vet Sci.  2010 Sep;11(3):205-211. 10.4142/jvs.2010.11.3.205.

Antigenic and immunogenic investigation of the virulence motif of the Newcastle disease virus fusion protein

• Choi KS
• Lee EK
• Jeon WJ
• Kwon JH

• KMID: 1093494
• DOI: http://doi.org/10.4142/jvs.2010.11.3.205

Abstract

Newcastle disease (ND) caused by virulent Newcastle disease virus (NDV) is a highly contagious viral disease of poultry. Virulent NDVs characteristically have a multibasic amino acid sequence (virulence motif) such as (112)RRQKRF(117) at the cleavage site of the precusor fusion (F0) protein. The antigenic and immunogenic characteristics of the virulence motif (112)RRQKRF(117) in the F0 protein of virulent NDVs were investigated. Epitope mapping analysis revealed that a RRQKRF-specific monoclonal antibody 4G2 recognized the KRF section of the motif. A synthetic peptide bearing the RRQKRF motif reacted strongly with sera from virulent NDV (with RRQKRF motif)-infected chickens. These sera also showed reactivity to peptides bearing other virulence motifs ((112)KRQKRF(117), (112)RRQRRF(117) and (112)RRRKRF(117)) but not an avirulence motif ((112)GRQGRL(117)) by ELISA. The synthetic bearing RRQKRF motif reacted with 60% to 91% of sera taken from surviving chickens on ND outbreak farms but not with sera from vaccinated birds, even though most of the sera had antibody to NDV due to vaccination. This indicates that the virulence motif has the potential to differentiate virulent NDV infected birds from vaccinated birds.

Keyword

B-cell epitope; fusion protein; Newcastle disease virus; virulence motif

MeSH Terms

Amino Acid Motifs/*immunology
Amino Acid Sequence
Animals
Chickens
Enzyme-Linked Immunosorbent Assay/veterinary
Epitope Mapping/veterinary
Newcastle Disease/*immunology
Newcastle disease virus/*genetics/pathogenicity
Poultry Diseases/*immunology/*virology
Serologic Tests/veterinary
Viral Fusion Proteins/*genetics/immunology
Virulence/genetics

Figure

• 

  Fig. 1 Fine mapping of the epitope within the virulence motif 112RRQKRF117 which is recognized by the anti-RRQKRF monoclonal antibody 4G2, using indirect ELISA. The BSA-conjugated peptides are designated in terms of the alanine substitution that was made, e.g. peptide R112A bears an arginine (R) to alanine (A) substitution at residue 112 of the fusion protein. BSA: bovine serum albumin.

• 

  Fig. 2 Reactivity of synthetic peptide RRQKRF to sera obtained from four chickens 0 and 7 days post-infection (dpi) with a virulent Newcastle disease virus (NDV) strain. Four (5-1, 5-2, 5-5 and 5-12) of 12 infected birds with virulent NDV survived 7 dpi. Surviving birds were tested 0 and 7 dpi. ELISA results of each serum tested was compared to HI titer (log2). Serum having PP value ≥ 20% in the pELISA was considered positive.

• 

  Fig. 3 Reactivity of anti-NDV chicken sera to synthetic peptides mimicking four virulence motifs (KRQKRF, RRQRRF and RRRKRF) and one avirulence motif (GRQGRL), as determined by indirect ELISA. These sera were obtained from four chickens 0 and 7 dpi after they were challenged with a virulent NDV strain.

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