Tembusu virus is a newly emerging flavivirus that caused egg-drop syndrome in ducks in China. TMUV envelope protein is a major structural protein locates at the surface of tembusu virus particle. During tembusu virus infection, envelope protein plays a pivotal role in induction of neutralizing antibody. However, B cell epitopes within envelope protein have not been well studied.
A series of 13 peptides derived from E protein of tembusu virus were synthesized and screened by Dot blot with tembusu virus-positive duck serum. Potential B-cell epitopes were respectively fused with GST tag and expressed in E. coli. The immunogenicity and protective efficiency of epitopes were assessed in ducks.
Dot blot assay identified the peptides P21 (amino acids 301–329), P23 (amino acids 369–387), P27 (amino acids 464–471) and P28 (amino acids 482–496) as potential B-cell epitopes within the envelope protein of tembusu virus. Immunization of prokaryotically expressed epitopes elicited specific antibodies in ducks and the specific antibody elicited by P21, P27 and P28 could neutralized tembusu virus. In addition, protective test suggested that P21 and P27 could completely protect immunized ducks from TMUV challenge.
Four potential B cell epiotpes within tembusu virus envelope protein were identified and analyzed in vitro and in vivo. It was demonstrated that two of them (P21 and P27) could elicit neutralizing antibodies in ducks and offer complete protection against tembusu virus challenge. This findings will contribute to the development of epitope vaccine for tembusu virus prevention.
Zhao D, Huang X, Liu Y, et al. Domain I and II from newly emerging goose tembusu virus envelope protein functions as a dominant-negative inhibitor of virus infectivity. Res Vet Sci. 2015;98:121–6. CrossRef
Su J, Li S, Hu X, et al. Duck egg-drop syndrome caused by BYD virus, a new Tembusu-related flavivirus. PLoS One. 2011;6(3):e18106. CrossRef
Zhang W, Chen S, Mahalingam S, et al. An updated review of avian-origin Tembusu virus: a newly emerging avian Flavivirus. J Gen Virol. 2017;98(10):2413–20. CrossRef
Wang Y, Yuan X, Li Y, et al. Rapid detection of newly isolated tembusu-related flavivirus by reverse-transcription loop-mediated isothermal amplification assay. Virol J. 2011;8:553. CrossRef
Ramanathan B, Poh CL, Kirk K, et al. Synthetic B-cell epitopes eliciting cross-neutralizing antibodies: strategies for future dengue vaccine. PLoS One. 2016;11(5):e0155900. CrossRef
Zhao D, Huang X, Han K, et al. Protective immune response against newly emerging goose tembusu virus infection induced by immunization with a recombinant envelope protein. Lett Appl Microbiol. 2015;61(4):318–24. CrossRef
Kuhn RJ, Zhang W, Rossmann MG, et al. Structure of dengue virus: implications for flavivirus organization, maturation, and fusion. Cell. 2002;108(5):717–25. CrossRef
Chu J, Rajamanonmani R, Li J, et al. Inhibition of West Nile virus entry by using a recombinant domain III from the envelope glycoprotein. J Gen Virol. 2005;86:405–12. CrossRef
Roehrig JT. Antigenic structure of flavivirus proteins. Adv Virus Res. 2003;59:141–75. CrossRef
Füzik T, Formanová P, Růžek D, et al. Structure of tick-borne encephalitis virus and its neutralization by a monoclonal antibody. Nat Commun. 2018;9(1):436. CrossRef
Rey FA, Heinz FX, Mandl C, et al. The envelope glycoprotein from tick-borne encephalitis virus at 2 a resolution. Nature. 1995;375(6529):291–8. CrossRef
Yu K, Sheng ZZ, Huang B, et al. Structural, antigenic, and evolutionary characterizations of the envelope protein of newly emerging duck Tembusu virus. PLoS One. 2013;8(8):e71319. CrossRef
Zu X, Liu Y, Wang S, et al. Peptide inhibitor of Japanese encephalitis virus infection targeting envelope protein domain III. Antivir Res. 2014;104:7–14. CrossRef
Zhang X, Jia R, Shen H, et al. Structures and functions of the envelope glycoprotein in flavivirus infections. Viruses. 2017;9(11). CrossRef
Wu HC, Jung MY, Chiu CY, et al. Identification of a dengue virus type 2 (DEN-2) serotype-specific B-cell epitope and detection of DEN-2-immunized animal serum samples using an epitope-based peptide antigen. J Gen Virol. 2003;84(Pt 10):2771–9. CrossRef
Oliphant T, Nybakken GE, Engle M, et al. Antibody recognition and neutralization determinants on domains I and II of West Nile virus envelope protein. J Virol. 2006;80(24):12149–59. CrossRef
Crill WD, Roehrig JT. Monoclonal antibodies that bind to domain III of dengue virus E glycoprotein are the most efficient blockers of virus adsorption to vero cells. J Virol. 2001;75:7769–73. CrossRef
Li C, Bai X, Meng R, et al. Identification of a new broadly cross-reactive epitope within domain III of the duck tembusu virus E protein. Sci Rep. 2016;6:36288. https://doi.org/10.1038/srep36288%20(2016).
Li C, Liu J, Shaozhou W, et al. Epitope identification and application for diagnosis of duck tembusu virus infections in ducks. Viruses. 2016;8(11). CrossRef
Huang X, Han K, Zhao D, et al. Identification and molecular characterization of a novel flavivirus isolated from geese in China. Res Vet Sci. 2013;94:774–80. CrossRef
Reed LJ, Muench H. A simple method of estimating fifty percent endpoints. Am J Hyg. 1938;27:493–7.
Zlatkovic J, Stiasny K, Heinz FX. Immunodominance and functional activities of antibody responses to inactivated West Nile virus and recombinant subunit vaccines in mice. J Virol. 2011;85(5):1994–2003. CrossRef
Zhang L, Li Z, Zhang Q, et al. Efficacy assessment of an inactivated Tembusu virus vaccine candidate in ducks. Res Vet Sci. 2017;110:72–8. CrossRef
Li W, Joshi MD, Singhania S, et al. Peptide vaccine: progress and challenges. Vaccines (Basel). 2014;2(3):515–36. CrossRef
Wei JC, Huang YZ, Zhong DK, et al. Design and evaluation of a multi-epitope peptide against Japanese encephalitis virus infection in BALB/c mice. Biochem Biophys Res Commun. 2010;396(4):787–92. CrossRef
Greenbaum JA, Andersen PH, Blythe M, et al. Towards a consensus on datasets and evaluation metrics for developing B-cell epitope prediction tools. J Mol Recognit. 2007;20(2):75–82. CrossRef
Van Regenmortel MH. Immunoinformatics may lead to a reappraisal of the nature of B cell epitopes and of the feasibility of synthetic peptide vaccines. J Mol Recognit. 2006;19(3):183–7. CrossRef
Song KY, Zhao H, Li SH, et al. Identification and characterization of a linearized B-cell epitope on the pr protein of dengue virus. J Gen Virol. 2013;94(Pt7):1510–6. CrossRef
Ti J, Li Z, Li X, et al. Identification of one B-cell epitope from NS1 protein of duck Tembusu virus with monoclonal antibodies. PLoS One. 2017;12(7):e0181177. CrossRef
Chen CW, Chang CY. Peptide scanning-assisted identification of a monoclonal antibody-recognized linear B-cell epitope. J Vis Exp. 2017;(121). https://doi.org/10.3791/55417.
Hughes HR, Crill WD, Davis BS, et al. A West Nile virus CD4 T cell epitope improves the immunogenicity of dengue virus serotype 2 vaccines. Virology. 2012;424(2):129–37. CrossRef
Chang GJ, Hunt AR, Holmes DA, et al. Enhancing biosynthesis and secretion of premembrane and envelope proteins by the chimeric plasmid of dengue virus type 2 and Japanese encephalitis virus. Virology. 2003;306(1):170–80. CrossRef
Yasri S, Wiwanitkit V. Finding of B cell epitope within polyprotein of Japanese encephalitis virus: a clue for development of new Japanese encephalitis vaccine. Arq Neuropsiquiatr. 2014;72(6):478. CrossRef
Mukhopadhyay S, Kuhn RJ, Rossmann MG. A structural perspective of the flavivirus life cycle. Nat Rev Microbiol. 2005;3(1):13–22. CrossRef
Puttikhunt C, Ong-Ajchaowlerd P, Prommool T, et al. Production and characterization of anti-dengue capsid antibodies suggesting the N terminus region covering the first 20 amino acids of dengue virus capsid protein is predominantly immunogenic in mice. Arch Virol. 2009;154(8):1211–121. CrossRef
Anandarao R, Swaminathan S, Khanna N. The identification of immunodominant linear epitopes of dengue type 2 virus capsid and NS4a proteins using pin-bound peptides. Virus Res. 2005;112(1–2):60–8. CrossRef
Lewis JK, Bothner B, Smith TJ, et al. Antiviral agent blocks breathing of the common cold virus. Proc Natl Acad Sci U S A. 1998;95(12):6774–8. CrossRef
Nadugala MN, Jeewandara C, Malavige GN, Premaratne PH, Goonasekara CL. Natural antibody responses to the capsid protein in sera of dengue infected patients from Sri Lanka. PLoS One. 2017;12(6):e0178009. CrossRef
- Screening and identification of B-cell epitopes within envelope protein of tembusu virus
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