Antigenic Analysis of Monoclonal Antibodies against Different Epitopes of σB Protein of Avian Reovirus

Abstract

BackgroundAvian reovirus (ARV) causes arthritis, tenosynovitis, runting-stunting syndrome (RSS), malabsorption syndrome (MAS) and immunosuppression in chickens. σB is one of the major structural proteins of ARV, which is able to induce group-specific antibodies against the virus.Methods and ResultsThe present study described the identification of two linear B-cell epitopes in ARV σB through expressing a set of partially overlapping and consecutive truncated peptides spanning σB screened with two monoclonal antibodies (mAbs) 1F4 and 1H3-1.The data indicated that 21KTPACW26 (epitope A) and 32WDTVTFH38 (epitope B) were minimal determinants of the linear B cell epitopes. Antibodies present in the serum of ARV-positive chickens recognized the minimal linear epitopes in Western blot analyses. By sequence alignment analysis, we determined that the epitopes A and B were not conserved among ARV, duck reovirus (DRV) and turkey reovirus (TRV) strains. Western blot assays, confirmed that epitopes A and B were ARV-specific epitopes, and they could not react with the corresponding peptides of DRV and TRV.Conclusions and SignificanceWe identified 21KTPACW26 and 32WDTVTFH38 as σB -specific epitopes recognized by mAbs 1F4 and 1H3-1, respectively. The results in this study may have potential applications in development of diagnostic techniques and epitope-based marker vaccines against ARV groups.

Development and Evaluation of a Monoclonal Antibody-Based Blocking Enzyme-Linked Immunosorbent Assay for the Detection of Antibodies against Novel Duck Reovirus in Waterfowl Species

“…The major outer capsid protein of NDRV is σB, which has antigenicity and can induce host-protective antibodies ( 15 , 16 , 30 , 31 ) and can also induce group-specific neutralizing antibody ( 16 , 30 , 32 ). It was previously reported that the most conserved amino acid in the deduced amino acid sequence of σB for avian orthoreovirus is in the N-terminal region (1 to 113 amino acids [aa]) ( 30 , 31 ). We developed a hybridoma that stably secreted MAb against NDRV and designated it 2-C10.…”

“…The VNT was performed in 96-well tissue culture microplates, as previously described, to determine the antibody specificities for diagnostic serum samples and quantitated antibody titers ( 30 ). In brief, the heat-inactivated sera were first diluted 5-fold and then serially diluted 2-fold (in duplicate) in 96-well plates with cell culture medium.…”

In vitro study on role of σB protein in avian reovirus pathogenesis

“…Most importantly s-class genome plays important role in inflammation, apoptosis and serodiagnosis. σB protein is found to be more conserved than other S-class proteins among ARV genes [ 38 ], but yet there is no report signifying the of role of σB in pathogenicity of ARV infection. This present in vitro study was carried out to examine pathogenesis related gene expression of host, in response to ARV infection and σB transfection.…”

Avian reovirus: a furious and fast evolving pathogen

“…However, the variability within genotypes is still high, and similar genotypes can also cause different pathology [27], so the molecular classification still needs to be improved. Moreover, other variable outer capsid proteins might play a role in ARV antigenicity, for example, lambda (λ)B and mu (µ)B [28,29], and should be considered Fig. 4.…”

“…However, the variability within genotypes is still high, and similar genotypes can also cause different pathology [27], so the molecular classification still needs to be improved. Moreover, other variable outer capsid proteins might play a role in ARV antigenicity, for example, lambda (λ)B and mu (µ)B [28, 29], and should be considered when typing strains. Finally, ARV is resistant to a wide range of heat, pH, and disinfectants, and virus particles can remain viable on wood shavings, feathers, shells, soil, water, feed, and other fomites for prolonged times [6], further hampering control efforts.…”