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Antigenic analysis of monoclonal antibodies against different epitopes of σB protein of avian reovirus.

Yin CH, Qin LT, Sun MY, Gao YL, Qi XL, Gao HL, Wang YQ, Wang XM - PLoS ONE (2013)

Bottom Line: 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.We identified (21)KTPACW(26) and (32)WDTVTFH(38) 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.

View Article: PubMed Central - PubMed

Affiliation: Division of Avian Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, P. R. China.

ABSTRACT

Background: Avian 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 results: The 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 (21)KTPACW(26) (epitope A) and (32)WDTVTFH(38) (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 significance: We identified (21)KTPACW(26) and (32)WDTVTFH(38) 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.

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Identification of cross-reactivity of mAb 1F4 or 1H3-1 to the associated coding motif of heterologous ARV, DRV and TRV strains.Marker: PageRulerTM Prestained Protein Ladder; GST-AΔNAQT containing the epitope A: KTPACW; M1-A: RSPACW; M2-A: RAPACW; M3-A: RTPACW; GST-BΔVPDVIRV containing the epitope B: WDTVTFH; M4--B: WDIEEFH; M5-B: WDSDIFH; M6-B: WNIETFH; M7-B: WDVETFH.
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pone-0081533-g006: Identification of cross-reactivity of mAb 1F4 or 1H3-1 to the associated coding motif of heterologous ARV, DRV and TRV strains.Marker: PageRulerTM Prestained Protein Ladder; GST-AΔNAQT containing the epitope A: KTPACW; M1-A: RSPACW; M2-A: RAPACW; M3-A: RTPACW; GST-BΔVPDVIRV containing the epitope B: WDTVTFH; M4--B: WDIEEFH; M5-B: WDSDIFH; M6-B: WNIETFH; M7-B: WDVETFH.

Mentions: To test the reactivity of mAbs 4F1 and 1H3 with corresponding motifs of DRV and TRV σB protein, two classes of peptides (M1-A, M2-A, M3-A; M4-B, M5-B, M6-B, M7-B) were expressed and then screened by Western blot. The results indicated that the mAb 1F4 could not react with M1-A, M2-A and M3-A, and the other mAb 1H3-1 could not recognize the recombinant fusion proteins of M4-B, M5-B, M6-B and M7-B, which suggested that epitopes A and B are not conserved in ARVs, DRVs and TRVs (Figure 6).


Antigenic analysis of monoclonal antibodies against different epitopes of σB protein of avian reovirus.

Yin CH, Qin LT, Sun MY, Gao YL, Qi XL, Gao HL, Wang YQ, Wang XM - PLoS ONE (2013)

Identification of cross-reactivity of mAb 1F4 or 1H3-1 to the associated coding motif of heterologous ARV, DRV and TRV strains.Marker: PageRulerTM Prestained Protein Ladder; GST-AΔNAQT containing the epitope A: KTPACW; M1-A: RSPACW; M2-A: RAPACW; M3-A: RTPACW; GST-BΔVPDVIRV containing the epitope B: WDTVTFH; M4--B: WDIEEFH; M5-B: WDSDIFH; M6-B: WNIETFH; M7-B: WDVETFH.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3842295&req=5

pone-0081533-g006: Identification of cross-reactivity of mAb 1F4 or 1H3-1 to the associated coding motif of heterologous ARV, DRV and TRV strains.Marker: PageRulerTM Prestained Protein Ladder; GST-AΔNAQT containing the epitope A: KTPACW; M1-A: RSPACW; M2-A: RAPACW; M3-A: RTPACW; GST-BΔVPDVIRV containing the epitope B: WDTVTFH; M4--B: WDIEEFH; M5-B: WDSDIFH; M6-B: WNIETFH; M7-B: WDVETFH.
Mentions: To test the reactivity of mAbs 4F1 and 1H3 with corresponding motifs of DRV and TRV σB protein, two classes of peptides (M1-A, M2-A, M3-A; M4-B, M5-B, M6-B, M7-B) were expressed and then screened by Western blot. The results indicated that the mAb 1F4 could not react with M1-A, M2-A and M3-A, and the other mAb 1H3-1 could not recognize the recombinant fusion proteins of M4-B, M5-B, M6-B and M7-B, which suggested that epitopes A and B are not conserved in ARVs, DRVs and TRVs (Figure 6).

Bottom Line: 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.We identified (21)KTPACW(26) and (32)WDTVTFH(38) 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.

View Article: PubMed Central - PubMed

Affiliation: Division of Avian Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, P. R. China.

ABSTRACT

Background: Avian 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 results: The 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 (21)KTPACW(26) (epitope A) and (32)WDTVTFH(38) (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 significance: We identified (21)KTPACW(26) and (32)WDTVTFH(38) 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.

Show MeSH
Related in: MedlinePlus