Skip to main content
Top
Published in: Virology Journal 1/2018

Open Access 01-12-2018 | Research

Screening and identification of B-cell epitopes within envelope protein of tembusu virus

Authors: Dongmin Zhao, Kaikai Han, Xinmei Huang, Lijiao Zhang, Huili Wang, Na Liu, Yujie Tian, Qingtao Liu, Jing Yang, Yuzhuo Liu, Yin Li

Published in: Virology Journal | Issue 1/2018

Login to get access

Abstract

Background

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.

Method

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.

Results

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.

Conclusion

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.
Literature
1.
go back to reference 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 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
2.
go back to reference Han K, Zhao D, Liu Y, et al. Quantitative proteomic analysis of duck ovarian follicles infected with duck tembusu virus by label-free LC-MS. Front Microbiol. 2016;7:463.PubMedPubMedCentral Han K, Zhao D, Liu Y, et al. Quantitative proteomic analysis of duck ovarian follicles infected with duck tembusu virus by label-free LC-MS. Front Microbiol. 2016;7:463.PubMedPubMedCentral
3.
go back to reference 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 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
4.
go back to reference 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 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
5.
go back to reference 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 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
6.
go back to reference 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 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
7.
go back to reference 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 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
8.
go back to reference 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 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
9.
go back to reference 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 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
10.
go back to reference Roehrig JT. Antigenic structure of flavivirus proteins. Adv Virus Res. 2003;59:141–75.CrossRef Roehrig JT. Antigenic structure of flavivirus proteins. Adv Virus Res. 2003;59:141–75.CrossRef
11.
go back to reference 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 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
12.
go back to reference 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 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
13.
go back to reference 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 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
14.
go back to reference 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 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
15.
go back to reference Zhang X, Jia R, Shen H, et al. Structures and functions of the envelope glycoprotein in flavivirus infections. Viruses. 2017;9(11).CrossRef Zhang X, Jia R, Shen H, et al. Structures and functions of the envelope glycoprotein in flavivirus infections. Viruses. 2017;9(11).CrossRef
16.
go back to reference 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 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
17.
go back to reference 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 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
18.
go back to reference 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 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
20.
go back to reference 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 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
21.
go back to reference 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 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
22.
go back to reference Reed LJ, Muench H. A simple method of estimating fifty percent endpoints. Am J Hyg. 1938;27:493–7. Reed LJ, Muench H. A simple method of estimating fifty percent endpoints. Am J Hyg. 1938;27:493–7.
23.
go back to reference 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 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
24.
go back to reference 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 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
25.
go back to reference Li W, Joshi MD, Singhania S, et al. Peptide vaccine: progress and challenges. Vaccines (Basel). 2014;2(3):515–36.CrossRef Li W, Joshi MD, Singhania S, et al. Peptide vaccine: progress and challenges. Vaccines (Basel). 2014;2(3):515–36.CrossRef
26.
go back to reference 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 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
27.
go back to reference 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 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
28.
go back to reference 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 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
29.
go back to reference 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 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
30.
go back to reference 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 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
32.
go back to reference 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 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
33.
go back to reference 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 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
34.
go back to reference 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 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
35.
go back to reference Mukhopadhyay S, Kuhn RJ, Rossmann MG. A structural perspective of the flavivirus life cycle. Nat Rev Microbiol. 2005;3(1):13–22.CrossRef Mukhopadhyay S, Kuhn RJ, Rossmann MG. A structural perspective of the flavivirus life cycle. Nat Rev Microbiol. 2005;3(1):13–22.CrossRef
36.
go back to reference 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 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
37.
go back to reference 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 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
38.
go back to reference 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 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
39.
go back to reference 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 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
Metadata
Title
Screening and identification of B-cell epitopes within envelope protein of tembusu virus
Authors
Dongmin Zhao
Kaikai Han
Xinmei Huang
Lijiao Zhang
Huili Wang
Na Liu
Yujie Tian
Qingtao Liu
Jing Yang
Yuzhuo Liu
Yin Li
Publication date
01-12-2018
Publisher
BioMed Central
Published in
Virology Journal / Issue 1/2018
Electronic ISSN: 1743-422X
DOI
https://doi.org/10.1186/s12985-018-1052-1

Other articles of this Issue 1/2018

Virology Journal 1/2018 Go to the issue