Characterization of Glycoproteins in Equine Herpesvirus-1

In this study, we attempted to express twelve glycoproteins of equine herpesvirus-1 (EHV-1) in 293T cells and to characterize these using monoclonal antibodies (MAbs) and horse sera against EHV-1. Expression of glycoprotein B (gB), gC, gD, gG, gI and gp2 was recognized by immunoblot analysis using h...

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Published inJournal of Veterinary Medical Science Vol. 75; no. 10; pp. 1317 - 1321
Main Authors Hassan Y. A. H. MAHMOUD, ANDOH, Kiyohiko, HATTORI, Shiho, TERADA, Yutaka, NOGUCHI, Keita, SHIMODA, Hiroshi, MAEDA, Ken
Format Journal Article
LanguageEnglish
Published Japan JAPANESE SOCIETY OF VETERINARY SCIENCE 2013
Japan Science and Technology Agency
The Japanese Society of Veterinary Science
Subjects
Animals
Antibodies, Monoclonal - immunology
Cell Line
equine herpesvirus-1
Female
glycoprotein
Glycoproteins - genetics
Glycoproteins - immunology
Herpesviridae Infections - immunology
Herpesviridae Infections - veterinary
Herpesviridae Infections - virology
Herpesvirus 1, Equid - genetics
Herpesvirus 1, Equid - immunology
Horse Diseases - immunology
Horse Diseases - virology
Horses
Immunoblotting - veterinary
Mice
Mice, Inbred BALB C
monoclonal antibody
Neutralization Tests - veterinary
Plasmids - genetics
Plasmids - immunology
Virology
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Abstract In this study, we attempted to express twelve glycoproteins of equine herpesvirus-1 (EHV-1) in 293T cells and to characterize these using monoclonal antibodies (MAbs) and horse sera against EHV-1. Expression of glycoprotein B (gB), gC, gD, gG, gI and gp2 was recognized by immunoblot analysis using horse sera, but that of gE, gH, gK, gL, gM and gN was not. Four MAbs recognized gB, four recognized gC and one recognized gp2. Two MAbs against gB cross-reacted with EHV-4. Interestingly, coexpression of gE and gI and gM and gN enhanced their antigenicity. Furthermore, immunoblot analysis of gp2 showed that different molecular masses of gp2 were recognized by the MAb against gp2 and horse sera against EHV-1. In this study, it was demonstrated that at least six glycoproteins were immunogenic to horses, and coexpression of gE and gI and gM and gN was important for enhancement of antigenicity.
AbstractList In this study, we attempted to express twelve glycoproteins of equine herpesvirus-1 (EHV-1) in 293T cells and to characterize these using monoclonal antibodies (MAbs) and horse sera against EHV-1. Expression of glycoprotein B (gB), gC, gD, gG, gI and gp2 was recognized by immunoblot analysis using horse sera, but that of gE, gH, gK, gL, gM and gN was not. Four MAbs recognized gB, four recognized gC and one recognized gp2. Two MAbs against gB cross-reacted with EHV-4. Interestingly, coexpression of gE and gI and gM and gN enhanced their antigenicity. Furthermore, immunoblot analysis of gp2 showed that different molecular masses of gp2 were recognized by the MAb against gp2 and horse sera against EHV-1. In this study, it was demonstrated that at least six glycoproteins were immunogenic to horses, and coexpression of gE and gI and gM and gN was important for enhancement of antigenicity.
In this study, we attempted to express twelve glycoproteins of equine herpesvirus-1 (EHV-1) in 293T cells and to characterize these using monoclonal antibodies (MAbs) and horse sera against EHV-1. Expression of glycoprotein B (gB), gC, gD, gG, gI and gp2 was recognized by immunoblot analysis using horse sera, but that of gE, gH, gK, gL, gM and gN was not. Four MAbs recognized gB, four recognized gC and one recognized gp2. Two MAbs against gB cross-reacted with EHV-4. Interestingly, coexpression of gE and gI and gM and gN enhanced their antigenicity. Furthermore, immunoblot analysis of gp2 showed that different molecular masses of gp2 were recognized by the MAb against gp2 and horse sera against EHV-1. In this study, it was demonstrated that at least six glycoproteins were immunogenic to horses, and coexpression of gE and gI and gM and gN was important for enhancement of antigenicity.
In this study, we attempted to express twelve glycoproteins of equine herpesvirus-1 (EHV-1) in 293T cells and to characterize these using monoclonal antibodies (MAbs) and horse sera against EHV-1. Expression of glycoprotein B (gB), gC, gD, gG, gI and gp2 was recognized by immunoblot analysis using horse sera, but that of gE, gH, gK, gL, gM and gN was not. Four MAbs recognized gB, four recognized gC and one recognized gp2. Two MAbs against gB cross-reacted with EHV-4. Interestingly, coexpression of gE and gI and gM and gN enhanced their antigenicity. Furthermore, immunoblot analysis of gp2 showed that different molecular masses of gp2 were recognized by the MAb against gp2 and horse sera against EHV-1. In this study, it was demonstrated that at least six glycoproteins were immunogenic to horses, and coexpression of gE and gI and gM and gN was important for enhancement of antigenicity.In this study, we attempted to express twelve glycoproteins of equine herpesvirus-1 (EHV-1) in 293T cells and to characterize these using monoclonal antibodies (MAbs) and horse sera against EHV-1. Expression of glycoprotein B (gB), gC, gD, gG, gI and gp2 was recognized by immunoblot analysis using horse sera, but that of gE, gH, gK, gL, gM and gN was not. Four MAbs recognized gB, four recognized gC and one recognized gp2. Two MAbs against gB cross-reacted with EHV-4. Interestingly, coexpression of gE and gI and gM and gN enhanced their antigenicity. Furthermore, immunoblot analysis of gp2 showed that different molecular masses of gp2 were recognized by the MAb against gp2 and horse sera against EHV-1. In this study, it was demonstrated that at least six glycoproteins were immunogenic to horses, and coexpression of gE and gI and gM and gN was important for enhancement of antigenicity.
Author TERADA, Yutaka
NOGUCHI, Keita
MAEDA, Ken
Hassan Y. A. H. MAHMOUD
ANDOH, Kiyohiko
HATTORI, Shiho
SHIMODA, Hiroshi
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  organization: Laboratory of Veterinary Microbiology, Joint Faculty of Veterinary Medicine, Yamaguchi University, 1677–1 Yoshida, Yamaguchi 753–8515, Japan
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  fullname: HATTORI, Shiho
  organization: Laboratory of Veterinary Microbiology, Joint Faculty of Veterinary Medicine, Yamaguchi University, 1677–1 Yoshida, Yamaguchi 753–8515, Japan
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  fullname: TERADA, Yutaka
  organization: Laboratory of Veterinary Microbiology, Joint Faculty of Veterinary Medicine, Yamaguchi University, 1677–1 Yoshida, Yamaguchi 753–8515, Japan
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  fullname: NOGUCHI, Keita
  organization: Laboratory of Veterinary Microbiology, Joint Faculty of Veterinary Medicine, Yamaguchi University, 1677–1 Yoshida, Yamaguchi 753–8515, Japan
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  fullname: SHIMODA, Hiroshi
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  fullname: MAEDA, Ken
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13. Farnsworth, A., Goldsmith, K. and Johnson, D. C. 2003. Herpes simplex virus glycoproteins gD and gE/gI serve essential but redundant functions during acquisition of the virion envelope in the cytoplasm. J. Virol. 77: 8481–8494.
6. Crabb, B. S., Allen, G. P. and Studdert, M. J. 1991. Characterization of the major glycoproteins of equine herpesviruses 4 and 1 and asinine herpesvirus 3 using monoclonal antibodies. J. Gen. Virol. 72: 2075–2082.
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References_xml – reference: 7. Crabb, B. S. and Studdert, M. J. 1993. Epitopes of glycoprotein G of equine herpesviruses 4 and 1 located near the C termini elicit type-specific antibody responses in the natural host. J. Virol. 67: 6332–6338.
– reference: 9. Crabb, B. S. and Studdert, M. J. 1995. Equine herpesviruses 4 (equine rhinpneumonitis virus) and 1(equine abortion virus). Adv. Virus Res. 45: 153–190.
– reference: 10. Crump, C. M., Bruun, B., Bell, S., Pomeranz, L. E., Minson, T. and Browne, H. M. 2004. Alphaherpesvirus glycoprotein M causes the relocalization of plasma membrane proteins. J. Gen. Virol. 85: 3517–3527.
– reference: 24. Studdert, M. J., Hartley, C. A., Dynon, K., Sandy, J. R., Slocombe, R. F., Charles, J. A., Milne, M. E., Clarke, A. F. and El-Hage, C. 2003. Outbreak of equine herpesvirus type 1 myeloencephalitis: new insights from virus identification by PCR and the application of an EHV-1-specific antibody detection ELISA. Vet. Rec. 153: 417–423.
– reference: 20. Paillot, R., Case, R., Ross, J., Newton, R. and Nugent, J. 2008. Equine herpes virus-1: virus, immunity and vaccines. Open Vet. Sci. J. 2: 68–91.
– reference: 22. Rudolph, J., Seyboldt, C., Granzow, H. and Osterrieder, N. 2002. The gene 10 (UL49.5) product of equine herpesvirus 1 is necessary and sufficient for functional processing of glycoprotein M. J. Virol. 76: 2952–2963.
– reference: 25. Sun, Y., MacLean, A. R., Dargan, D. and Brown, S. M. 1994. Identification and characterization of the protein product of gene 71 in equine herpesvirus1. J. Gen. Virol. 75: 3117–3126.
– reference: 16. Learmonth, G. S., Love, D. N., Wellington, J. E., Gilkerson, J. R. and Whalley, J. M. 2002. The C-terminal regions of the envelope glycoprotein gp2 of equine herpesviruses 1 and 4 are antigenically distinct. Arch. Virol. 147: 607–615.
– reference: 1. Allen, G. P. and Bryans, J. T. 1986. Molecular epizootiology, pathogenesis, and prophylaxis of equine herpesvirus-1 infections. Prog. Vet. Microbiol. Immunol. 2: 78–144.
– reference: 26. Telford, E. A., Watson, M. S., McBride, K. and Davison, A. J. 1992. The DNA sequence of equine herpesvirus-1. Virology 189: 304–316.
– reference: 17. Maeda, K., Yasumoto, S., Tsuruda, A., Andoh, K., Kai, K., Otoi, T. and Matsumura, T. 2007. Establishment of a novel equine cell line for isolation and propagation of equine herpesviruses. J. Vet. Med. Sci. 69: 989–991.
– reference: 28. Turtinen, L. W. and Allen, G. P. 1982. Identification of the envelope surface glycoproteins of equine herpesvirus type 1. J. Gen. Virol. 63: 481–485.
– reference: 19. Norrild, B. 1985. Humoral responses to herpes simplex virus infections. pp. 69–86. In: The Herpesviruses 4 (Roizman, B. and Lopez, C. eds.), Plenum Press, New York.
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Snippet In this study, we attempted to express twelve glycoproteins of equine herpesvirus-1 (EHV-1) in 293T cells and to characterize these using monoclonal antibodies...
In this study, we attempted to express twelve glycoproteins of equine herpesvirus-1 (EHV-1) in 293T cells and to characterize these using monoclonal antibodies...
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SubjectTerms Animals
Antibodies, Monoclonal - immunology
Cell Line
equine herpesvirus-1
Female
glycoprotein
Glycoproteins - genetics
Glycoproteins - immunology
Herpesviridae Infections - immunology
Herpesviridae Infections - veterinary
Herpesviridae Infections - virology
Herpesvirus 1, Equid - genetics
Herpesvirus 1, Equid - immunology
Horse Diseases - immunology
Horse Diseases - virology
Horses
Immunoblotting - veterinary
Mice
Mice, Inbred BALB C
monoclonal antibody
Neutralization Tests - veterinary
Plasmids - genetics
Plasmids - immunology
Virology
Title Characterization of Glycoproteins in Equine Herpesvirus-1
URI https://www.jstage.jst.go.jp/article/jvms/75/10/75_13-0168/_article/-char/en
https://www.ncbi.nlm.nih.gov/pubmed/23748975
https://www.proquest.com/docview/1478009249
https://www.proquest.com/docview/1448215973
https://pubmed.ncbi.nlm.nih.gov/PMC3942931
Volume 75
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