Matrix (M) protein of Newcastle disease virus (NDV) is an abundant protein that can induce a robust humoral immune response. However, its antigenic epitopes remain unknown. In this study, we used a pepscan approach to map linear B cell immunodominant epitopes (IDEs) of M protein with NDV-specific chicken antisera. The six epitopes with the highest reactivity by peptide scanning were obtained as IDE candidates. Among them, aa71–85 and aa349–363 were identified by immunological assays with NDV-specific or IDE-specific antisera. The minimal antigenic epitopes of the two IDEs were further characterized as 77MIDDKP82 and 354HTLAKYNPFK363. Moreover, an amino acid sequence alignment and immunoblot analysis revealed the conservation of the two IDEs in the M protein of strains of different genotypes. These two IDEs of M protein could be genetically eliminated as negative markers in recombinant NDV for serologically differential diagnosis in the development of marker vaccines.
Get full access to this article
View all access options for this article.
References
1.
AldousEW, and AlexanderDJ. Detection and differentiation of Newcastle disease virus (avian paramyxovirus type 1). Avian Pathol, 2001; 30:117–128.
2.
BattistiAJ, MengG, WinklerDC, et al.Structure and assembly of a paramyxovirus matrix protein. Proc Natl Acad Sci U S A, 2012; 109:13996–14000.
3.
ChambersP, and SamsonAC. A new structural protein for Newcastle disease virus. J Gen Virol, 1980; 50:155–166.
4.
ChenY, ZhangJ, QiaoC, et al.Identification of a linear epitope on the haemagglutinin protein of pandemic a/h1n1 2009 influenza virus using monoclonal antibodies. Arch Virol, 2014; 159:1413–1419.
5.
ChoSH, KwonHJ, KimTE, et al.Characterization of a recombinant Newcastle disease virus vaccine strain. Clin Vaccine Immunol, 2008; 15:1572–1579.
6.
ColemanNA, and PeeplesME. The matrix protein of Newcastle disease virus localizes to the nucleus via a bipartite nuclear localization signal. Virology, 1993; 195:596–607.
7.
DortmansJC, RottierPJ, KochG, et al.The viral replication complex is associated with the virulence of Newcastle disease virus. J Virol, 2010; 84:10113–10120.
8.
DuanZ, HuZ, ZhuJ, et al.Mutations in the FPIV motif of Newcastle disease virus matrix protein attenuate virus replication and reduce virus budding. Arch Virol, 2014; 159:1813–1819.
9.
DuanZ, LiJ, ZhuJ, et al.A single amino acid mutation, R42A, in the Newcastle disease virus matrix protein abrogates its nuclear localization and attenuates viral replication and pathogenicity. J Gen Virol, 2014; 95:1067–1073.
10.
DuanZ, SongQ, WangY, et al.Characterization of signal sequences determining the nuclear export of Newcastle disease virus matrix protein. Arch Virol, 2013; 158:2589–2595.
11.
DuanZ, XuH, JiX, et al.Importin α5 negatively regulates importin β1-mediated nuclear import of Newcastle disease virus matrix protein and viral replication and pathogenicity in chicken fibroblasts. Virulence, 2018; 9:783–803.
12.
GeysenHM, MeloenRH, and BartelingSJ. Use of peptide synthesis to probe viral antigens for epitopes to a resolution of a single amino acid. Proc. Natl Acad Sci U S A, 1984; 81:3996–4000.
13.
GhildyalR, HoA, WagstaffKM, et al.Nuclear import of the respiratory syncytial virus matrix protein is mediated by importin beta1 independent of importin alpha. Biochemistry, 2005; 44:12887–12895.
14.
GlodowskiDR, PetersenJM, and DahlbergJE. Complex nuclear localization signals in the matrix protein of vesicular stomatitis virus. J Biol Chem, 2002; 277:46864–46870.
15.
GommePT, StantonPG, and HearnMT. Evaluation of a pepscan approach to identify epitopes recognised by anti-htsh monoclonal antibodies. J Biochem Biophys Methods, 1999; 38:53–70.
16.
HarrisonMS, SakaguchiT, and SchmittAP. Paramyxovirus assembly and budding: building particles that transmit infections. Int J Biochem Cell Biol, 2010; 42:1416–1429.
17.
IramN, ShahMS, and IsmatF. Heterologous expression, characterization and evaluation of the matrix protein from Newcastle disease virus as a target for antiviral therapies. Appl Microbiol Biotechnol, 2014; 984:1691–1701.
18.
JespersenMC, PetersB, NielsenM, et al.Bepipred-2.0: improving sequence-based B-cell epitope prediction using conformational epitopes. Nucleic Acids Res, 2017; 45:24–29.
19.
LangedijkJP, MeloenRH, and van OirschotJT. Identification of a conserved neutralization site in the first heptad repeat of the fusion protein of respiratory syncytial virus. Arch Virol, 1998; 143:313–320.
20.
LanzavecchiaA, FrühwirthA, PerezL, et al.Antibody-guided vaccine design: identification of protective epitopes. Curr Opin Immunol, 2016; 41:62–67.
21.
LinM, McraeH, DanHH, et al.High-resolution epitope mapping for monoclonal antibodies to the structural protein erns of classical swine fever virus using peptide array and random peptide phage display approaches. J Gen Virol, 2010; 91:2928–2940.
22.
Loomis-PriceLD. Linear epitope mapping by the pepscan method. Methods Mol Med, 1999; 17:293–307.
23.
MebatsionT, KoolenMJ, de VaanLT, et al.Newcastle disease virus (NDV) marker vaccine: an immunodominant epitope on the nucleoprotein gene of NDV can be deleted or replaced by a foreign epitope. J Virol, 2002; 76:10138–10146.
24.
MillerPJ, DecaniniEL, and AfonsoCL. Newcastle disease: evolution of genotypes and the related diagnostic challenges. Infect Genet Evol, 2010; 10:26–35.
25.
PanshinA, ShihmanterE, WeismanY, et al.Antigenic epitope characterization of matrix protein of Newcastle disease virus using monoclonal antibody approach: contrasting variability amongst NDV strains. Comp Immunol Microbiol Infect Dis, 1997; 20:177–189.
26.
PantuaHD, McginnesLW, PeeplesME, et al.Requirements for the assembly and release of Newcastle disease virus-like particles. J Virol, 2006; 80:11062–11073.
27.
PeeplesME, WangC, GuptaKC, et al.Nuclear entry and nucleolar localization of the Newcastle disease virus (NDV) matrix protein occur early in infection and do not require other NDV proteins. J Virol, 1992; 66:3263–3269.
28.
QiG, LinJ, CaoR, et al.Asparaginase display of polypeptides in the periplasm of escherichia coli: potential rapid pepscan technique for antigen epitope mapping. J Immunol Methods, 2005; 299:9–19.
ReynoldsDL, and MaraqaAD. Protective immunity against Newcastle disease: the role of antibodies specific to Newcastle disease virus polypeptides. Avian Dis, 2000; 44:138–144.
31.
RoggenEL. B-cell epitope engineering: a matter of recognizing protein features and motives. Drug Discov Today Technol, 2008; 5:e49–e55.
32.
SealBS, KingDJ, and MeinersmannRJ. Molecular evolution of the Newcastle disease virus matrix protein gene and phylogenetic relationships among the paramyxoviridae. Virus Res, 2000; 66:1–11.
33.
SetteA, and FikesJ. Epitope-based vaccines: an update on epitope identification, vaccine design and delivery. Curr Opin Immunol, 2003; 15:461–470.
34.
SuckauD, KöhlJ, KarwathG, et al.Molecular epitope identification by limited proteolysis of an immobilized antigen-antibody complex and mass spectrometric peptide mapping. Proc Natl Acad Sci U S A, 1990; 87:9848–9852.
35.
WangX, QuiL, DangY, et al.Linear epitope recognition antibodies strongly respond to the C-terminal domain of HP-PRRSV GP5. Vet Microbiol, 2014; 174:565–569.
36.
WestoverKM, and HughesAL. Molecular evolution of viral fusion and matrix protein genes and phylogenetic relationships among the paramyxoviridae. Mol Phylogenet Evol, 2001; 21:128–134.
37.
YusoffK, and TanWS. Newcastle disease virus: macromolecules and opportunities. Avian Pathol, 2001; 30:439–455.
38.
ZanettiF, RodriguezM, KingDJ, et al.Matrix protein gene sequence analysis of avian paramyxovirus 1 isolates obtained from pigeons. Virus Genes, 2003; 26:199–206.
39.
ZhaoS, QiT, GuoW, et al.Identification of a conserved B-cell epitope in the equine arteritis virus (EAV) N protein using the pepscan technique. Virus Genes, 2013; 47:292–297.
Supplementary Material
Please find the following supplemental material available below.
For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.
For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.
