Please wait a minute...
Shopping Cart Email List Chinese
Advanced Search Fig/Tab
Frontiers of Agricultural Science and Engineering

ISSN 2095-7505

ISSN 2095-977X(Online)

CN 10-1204/S

Postal Subscription Code 80-906

Featured Articles  |  Most Downloaded  |  Most Reading
Online Submission Subscribe to Our RSS Content Feed
Front. Agr. Sci. Eng.    2015, Vol. 2 Issue (3) : 216-222    https://doi.org/10.15302/J-FASE-2015072
RESEARCH ARTICLE
Research and development of a novel subunit vaccine for the currently circulating pseudorabies virus variant in China
Yuzhou WANG1,Tongyan WANG1,He YAN1,Fanli YANG1,Linghua GUO1,Qingyuan YANG1,Xule HU1,Feifei TAN1,Yan XIAO1,Xiangdong LI1,*(),Kegong TIAN1,2,*()
1. National Research Center for Veterinary Medicine, Luoyang 471003, China
2. College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zhengzhou 450002, China
 Download: PDF(827 KB)   HTML
 Export: BibTeX | EndNote | Reference Manager | ProCite | RefWorks
Abstract

Pseudorabies (PR) is a devastating viral disease which leads to fatal encephalitis and respiratory disorders in pigs. Commercial gE-deleted live pseudorabies virus (PRV) vaccine has been widely used to control this disease in China. However, the new-emerging variants of PRV compromises the protection provided by current vaccines and lead to the outbreak of PR in vaccinated pig herds. Several killed and live vaccine candidates based on current PRV variants have been reported to be effective to control the disease. A subunit vaccine based on gB protein, one major PRV glycoprotein which elicits strong humoral and cellular immune responses, however, was never evaluated for protection against the current circulating PRV variants. In this study, full-length PRV gB protein was successfully expressed in baculovirus/insect cells in the soluble format and was tested on 3-week-old piglets as a subunit vaccine. Compared with unvaccinated pigs, the gB-vaccinated pigs developed specific antibody-mediated responses and were protected from the virulent PRV HN1201 challenge. All vaccinated pigs survived without showing any PRV-specific respiratory and neurological signs, but all unvaccinated pigs died within 7 days after HN1201 challenge. Hence, this novel gB-based vaccine could be applied as an effective subunit vaccine to control PRV variant in China.
Keywords pseudorabies virus      glycoprotein B protein      subunit vaccine     
Corresponding Author(s): Xiangdong LI,Kegong TIAN   
Just Accepted Date: 22 October 2015   Online First Date: 03 November 2015    Issue Date: 10 November 2015
 Cite this article:   
Yuzhou WANG,Tongyan WANG,He YAN, et al. Research and development of a novel subunit vaccine for the currently circulating pseudorabies virus variant in China[J]. Front. Agr. Sci. Eng. , 2015, 2(3): 216-222.
 URL:  
https://academic.hep.com.cn/fase/EN/10.15302/J-FASE-2015072
https://academic.hep.com.cn/fase/EN/Y2015/V2/I3/216
Fig.1  Expression and purification of gB protein analysis by SDS-PAGE (a) and western blot (b). (a) Lane 1, the supernatant of sf9 insect cell culture; lane 2, purified S1 protein using immunoaffinity chromatography; lane 3, protein marker; (b) lane 1, protein marker; lane 2, the recombinant protein was recognized by HIS-tag monoclonal antibody; lane 3, the recombinant protein was recognized by PRV-gB specific monoclonal antibody.
Fig.2  Body temperature (a) and clinical symptom score (b) of pigs within first 7 days after PRV HN1201 challenge. *P<0.05.
Fig.3  Body weight gain (a) and survival rate of pigs (b) after PRV HN1201 challenge. *P<0.05.
Fig.4  Profile of PRV gB-specific (a) and gE-specific (b) antibody responses after vaccination and challenge.
Fig.5  Histopathological results of vaccinated (a−e) and unvaccinated pigs (f−j). (a−e) Tonsil, lung, brain, cerebellum and trigeminal ganglia samples from vaccinated pigs, respectively. No obvious histopathological changes were observed; (f) tonsil. Tonsillar lymphoid tissue necrosis and formation of large necrotic foci; (g) lung. Massive vascular congestion and hemorrhage; (h) brain. Nonsuppurative meningoencephalitis; (i) cerebellum. Purkinje cell degeneration and necrosis; (j) trigeminal ganglia. Infiltration of lymphocytes around the neuron of trigeminal ganglia. Original magnification 400×.
Fig.6  Immunochemistry staining of tonsil (a, f), lung (b, g), brain (c, h), cerebellum (d, i), and trigeminal ganglia (e, j) samples of vaccinated (a−e) and unvaccinated pigs (f−j). Original magnification 400×.
GroupsTonsilLungBrainCerebellumTrigeminal ganglia
DMEM++–++-+
+++++++++++
++++++++++
–+ +–++
+++++––+
Vaccine–––––
––––+
––––+
––––+
–––––
Tab.1  Virus antigen distribution and intensity in different organs detected by immunohistochemistry staining
1 Wittmann  G, Rziha  H J. Aujeszky’s disease (pseudorabies) in pigs. Herpesvirus Diseases of Cattle, Horses, and Pigs. New York: Springer, 1989: 230–325
2 An  T Q, Peng  J M, Tian  Z J, Zhao  H Y, Li  N, Liu  Y M, Chen  J Z, Leng  C L, Sun  Y, Chang  D, Tong  G Z. Pseudorabies virus variant in Bartha-K61-vaccinated pigs, China, 2012. Emerging Infectious Diseases, 2013, 19(11): 1749–1755
https://doi.org/10.3201/eid1911.130177 pmid: 24188614
3 Luo  Y, Li  N, Cong  X, Wang  C H, Du  M, Li  L, Zhao  B, Yuan  J, Liu  D D, Li  S, Li  Y, Sun  Y, Qiu  H J. Pathogenicity and genomic characterization of a pseudorabies virus variant isolated from Bartha-K61-vaccinated swine population in China. Veterinary Microbiology, 2014, 174(1−2): 107–115
https://doi.org/10.1016/j.vetmic.2014.09.003 pmid: 25293398
4 Yu  X, Zhou  Z, Hu  D, Zhang  Q, Han  T, Li  X, Gu  X, Yuan  L, Zhang  S, Wang  B, Qu  P, Liu  J, Zhai  X, Tian  K. Pathogenic pseudorabies virus, China, 2012. Emerging Infectious Diseases, 2014, 20(1): 102–104
https://doi.org/10.3201/eid2001.130531 pmid: 24377462
5 Wu  R, Bai  C, Sun  J, Chang  S, Zhang  X. Emergence of virulent pseudorabies virus infection in northern China. Journal of Veterinary Science, 2013, 14(3): 363–365
https://doi.org/10.4142/jvs.2013.14.3.363 pmid: 23820207
6 Wang  C H, Yuan  J, Qin  H Y, Luo  Y, Cong  X, Li  Y, Chen  J, Li  S, Sun  Y, Qiu  H J. A novel gE-deleted pseudorabies virus (PRV) provides rapid and complete protection from lethal challenge with the PRV variant emerging in Bartha-K61-vaccinated swine population in China. Vaccine, 2014, 32(27): 3379–3385
https://doi.org/10.1016/j.vaccine.2014.04.035 pmid: 24793946
7 Gu  Z, Dong  J, Wang  J, Hou  C, Sun  H, Yang  W, Bai  J, Jiang  P. A novel inactivated gE/gI deleted pseudorabies virus (PRV) vaccine completely protects pigs from an emerged variant PRV challenge. Virus Research, 2015, 195: 57–63
https://doi.org/10.1016/j.virusres.2014.09.003 pmid: 25240533
8 Zhang  C, Guo  L, Jia  X, Wang  T, Wang  J, Sun  Z, Wang  L, Li  X, Tan  F, Tian  K. Construction of a triple gene-deleted Chinese Pseudorabies virus variant and its efficacy study as a vaccine candidate on suckling piglets. Vaccine, 2015, 33(21): 2432–2437
https://doi.org/10.1016/j.vaccine.2015.03.094 pmid: 25865469
9 Klupp  B G, Hengartner  C J, Mettenleiter  T C, Enquist  L W. Complete, annotated sequence of the pseudorabies virus genome. Journal of Virology, 2004, 78(1): 424–440
https://doi.org/10.1128/JVI.78.1.424-440.2004 pmid: 14671123
10 Mulder  W A, Pol  J M, Gruys  E, Jacobs  L, De Jong  M C, Peeters  B P, Kimman  T G. Pseudorabies virus infections in pigs. Role of viral proteins in virulence, pathogenesis and transmission. Veterinary Research, 1997, 28(1): 1–17
pmid: 9172836
11 Han  Y W, Aleyas  A G, George  J A, Kim  S J, Kim  H K, Yoon  H A, Yoo  D J, Kang  S H, Kim  K, Eo  S K. Polarization of protective immunity induced by replication-incompetent adenovirus expressing glycoproteins of pseudorabies virus. Experimental & Molecular Medicine, 2008, 40(6): 583–595
https://doi.org/10.3858/emm.2008.40.6.583 pmid: 19116444
12 van Rooij  E M, Haagmans  B L, Glansbeek  H L, de Visser  Y E, de Bruin  M G, Boersma  W, Bianchi  A T. A DNA vaccine coding for glycoprotein B of pseudorabies virus induces cell-mediated immunity in pigs and reduces virus excretion early after infection. Veterinary Immunology and Immunopathology, 2000, 74(1−2): 121–136
https://doi.org/10.1016/S0165-2427(00)00170-7 pmid: 10760394
13 Grabowska  A K, Lipińska  A D, Rohde  J, Szewczyk  B, Bienkowska-Szewczyk  K, Rziha  H J. New baculovirus recombinants expressing Pseudorabies virus (PRV) glycoproteins protect mice against lethal challenge infection. Vaccine, 2009, 27(27): 3584–3591
https://doi.org/10.1016/j.vaccine.2009.03.067 pmid: 19464538
14 Kim  S J, Kim  H K, Han  Y W, Aleyas  A G, George  J A, Yoon  H A, Yoo  D J, Kim  K, Eo  S K. Multiple alternating immunizations with DNA vaccine and replication incompetent adenovirus expressing gB of pseudorabies virus protect animals against lethal virus challenge. Journal of Microbiology and Biotechnology, 2008, 18(7): 1326–1334
pmid: 18667863
15 Gerdts  V, Jöns  A, Mettenleiter  T C. Potency of an experimental DNA vaccine against Aujeszky’s disease in pigs. Veterinary Microbiology, 1999, 66(1): 1–13
https://doi.org/10.1016/S0378-1135(98)00300-9 pmid: 10223318
16 Miller  L C, Zanella  E L, Waters  W R, Lager  K M. Cytokine protein expression levels in tracheobronchial lymph node homogenates of pigs infected with pseudorabies virus. Clinical and Vaccine Immunology, 2010, 17(5): 728–734
https://doi.org/10.1128/CVI.00485-09 pmid: 20219878
17 Wang  T, Xiao  Y, Yang  Q, Wang  Y, Sun  Z, Zhang  C, Yan  S, Wang  J, Guo  L, Yan  H, Gao  Z, Wang  L, Li  X, Tan  F, Tian  K. Construction of a gE-deleted pseudorabies virus and its efficacy to the new-emerging variant PRV challenge in the form of killed vaccine. BioMed Research International, 2015, 2015: 684945
pmid: 26457302
18 Dory  D, Torché  A M, Béven  V, Blanchard  P, Loizel  C, Cariolet  R, Jestin  A. Effective protection of pigs against lethal Pseudorabies virus infection after a single injection of low-dose Sindbis-derived plasmids encoding PrV gB, gC and gD glycoproteins. Vaccine, 2005, 23(26): 3483–3491
https://doi.org/10.1016/j.vaccine.2004.10.050 pmid: 15837372
19 Mettenleiter  T C. Immunobiology of pseudorabies (Aujeszky’s disease). Veterinary Immunology and Immunopathology, 1996, 54(1−4): 221–229
https://doi.org/10.1016/S0165-2427(96)05695-4 pmid: 8988868
20 Mettenleiter  T C. Pseudorabies (Aujeszky’s disease) virus: state of the art. August 1993. Acta Veterinaria Hungarica, 1994, 42(2−3): 153–177
pmid: 7810409
[1] Shijun YAN,He YAN,Chaolin ZHANG,Tongyan WANG,Qingyuan YANG,Zhe SUN,Yan XIAO,Feifei TAN,Xiangdong LI,Kegong TIAN. RR1 and RR2 gene deletion affects the immunogenicity of a live attenuated pseudorabies virus vaccine candidate in natural pig host[J]. Front. Agr. Sci. Eng. , 2016, 3(1): 81-86.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed