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Analysis of murine B-cell epitopes on Eastern equine encephalitis virus glycoprotein E2

Applied Microbiology and Biotechnology volume 97, pages 6359–6372 (2013)Cite this article

Abstract

The Eastern equine encephalitis virus (EEEV) E2 protein is one of the main targets of the protective immune response against EEEV. Although some efforts have done to elaborate the structure and immune molecular basis of Alphaviruses E2 protein, the published data of EEEV E2 are limited. Preparation of EEEV E2 protein-specific antibodies and define MAbs-binding epitopes on E2 protein will be conductive to the antibody-based prophylactic and therapeutic and to the study on structure and function of EEEV E2 protein. In this study, 51 EEEV E2 protein-reactive monoclonal antibodies (MAbs) and antisera (polyclonal antibodies, PAbs) were prepared and characterized. By pepscan with MAbs and PAbs using enzyme-linked immunosorbent assay, we defined 18 murine linear B-cell epitopes. Seven peptide epitopes were recognized by both MAbs and PAbs, nine epitopes were only recognized by PAbs, and two epitopes were only recognized by MAbs. Among the epitopes recognized by MAbs, seven epitopes were found only in EEEV and two epitopes were found both in EEEV and Venezuelan equine encephalitis virus (VEEV). Four of the EEEV antigenic complex-specific epitopes were commonly held by EEEV subtypes I/II/III/IV (1-16aa, 248-259aa, 271-286aa, 321-336aa probably located in E2 domain A, domain B, domain C, domain C, respectively). The remaining three epitopes were EEEV type-specific epitopes: a subtype I-specific epitope at amino acids 108–119 (domain A), a subtype I/IV-specific epitope at amino acids 211–226 (domain B) and a subtype I/II/III-specific epitope at amino acids 231–246 (domain B). The two common epitopes of EEEV and VEEV were located at amino acids 131–146 and 241–256 (domain B). The generation of EEEV E2-specific MAbs with defined specificities and binding epitopes will inform the development of differential diagnostic approaches and structure study for EEEV and associated alphaviruses.

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Acknowledgments

The authors thank Dr. Peter Wilker for editing the manuscript. This study was supported by National High-Tech Research and Development Program of China (no. 2011AA10A212) and grants from State Key Laboratory of Veterinary Biotechnology (SKLVBP201018 and SKLVBP201210).

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  1. The Key Laboratory of Veterinary Public Health, Ministry of Agriculture, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, 427 Maduan Street, Nangang District, Harbin, 150001, People’s Republic of China

    Sun EnCheng, Zhao Jing, Yang Tao, Xu QingYuan, Qin YongLi, Wang WenShi, Wei Peng, Sun Liang, Sun Jing & Wu DongLai

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  1. Sun EnCheng
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  2. Zhao Jing
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  3. Yang Tao
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Correspondence to Wu DongLai.

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EnCheng, S., Jing, Z., Tao, Y. et al. Analysis of murine B-cell epitopes on Eastern equine encephalitis virus glycoprotein E2. Appl Microbiol Biotechnol 97, 6359–6372 (2013). https://doi.org/10.1007/s00253-013-4819-8

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  • DOI: https://doi.org/10.1007/s00253-013-4819-8

Keywords

  • Eastern equine encephalitis virus
  • E2 protein
  • B-cell epitope mapping