Abstract
We recently established an in vitro model of equine arteritis virus (EAV) persistence in HeLa cells. The objective of this study was to determine whether viral variants with novel neutralization phenotypes emerged during persistent EAV infection of HeLa cells, as occurs during viral persistence in carrier stallions. Viruses recovered from persistently infected HeLa cells had different neutralization phenotypes than the virus in the original inoculum, as determined by neutralization assays using EAV-specific monoclonal antibodies and polyclonal equine antisera raised against different strains of EAV. Comparative sequence analyses of the entire structural protein genes (ORFs 2a, 2b, and 3-7) of these viruses, coupled with construction of chimeric viruses utilizing an infectious cDNA clone of EAV, confirmed that the alterations in neutralization phenotype were caused by amino acid changes in the GP5 protein encoded by ORF5. Site-directed mutagenesis studies unequivocally confirmed that amino acid 98 in the GP5 protein was responsible for the altered neutralization phenotype of these viruses. Amino acid 98 in the GP5 protein, which has not previously been identified as a neutralization determinant of EAV, should be included in an expanded neutralization site D (amino acids 98-106).
| Original language | English (US) |
|---|---|
| Pages (from-to) | 150-153 |
| Number of pages | 4 |
| Journal | Virus Research |
| Volume | 138 |
| Issue number | 1-2 |
| DOIs | |
| State | Published - Dec 2008 |
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Keywords
- Equine arteritis virus
- HeLa cells
- Neutralization determinant
- Persistent infection
ASJC Scopus subject areas
- Virology
- Infectious Diseases
- Cancer Research
Cite this
Identification of an additional neutralization determinant of equine arteritis virus. / Zhang, Jianqiang; Timoney, Peter J.; Maclachlan, Nigel J; Balasuriya, Udeni B R.
In: Virus Research, Vol. 138, No. 1-2, 12.2008, p. 150-153.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Identification of an additional neutralization determinant of equine arteritis virus
AU - Zhang, Jianqiang
AU - Timoney, Peter J.
AU - Maclachlan, Nigel J
AU - Balasuriya, Udeni B R
PY - 2008/12
Y1 - 2008/12
N2 - We recently established an in vitro model of equine arteritis virus (EAV) persistence in HeLa cells. The objective of this study was to determine whether viral variants with novel neutralization phenotypes emerged during persistent EAV infection of HeLa cells, as occurs during viral persistence in carrier stallions. Viruses recovered from persistently infected HeLa cells had different neutralization phenotypes than the virus in the original inoculum, as determined by neutralization assays using EAV-specific monoclonal antibodies and polyclonal equine antisera raised against different strains of EAV. Comparative sequence analyses of the entire structural protein genes (ORFs 2a, 2b, and 3-7) of these viruses, coupled with construction of chimeric viruses utilizing an infectious cDNA clone of EAV, confirmed that the alterations in neutralization phenotype were caused by amino acid changes in the GP5 protein encoded by ORF5. Site-directed mutagenesis studies unequivocally confirmed that amino acid 98 in the GP5 protein was responsible for the altered neutralization phenotype of these viruses. Amino acid 98 in the GP5 protein, which has not previously been identified as a neutralization determinant of EAV, should be included in an expanded neutralization site D (amino acids 98-106).
AB - We recently established an in vitro model of equine arteritis virus (EAV) persistence in HeLa cells. The objective of this study was to determine whether viral variants with novel neutralization phenotypes emerged during persistent EAV infection of HeLa cells, as occurs during viral persistence in carrier stallions. Viruses recovered from persistently infected HeLa cells had different neutralization phenotypes than the virus in the original inoculum, as determined by neutralization assays using EAV-specific monoclonal antibodies and polyclonal equine antisera raised against different strains of EAV. Comparative sequence analyses of the entire structural protein genes (ORFs 2a, 2b, and 3-7) of these viruses, coupled with construction of chimeric viruses utilizing an infectious cDNA clone of EAV, confirmed that the alterations in neutralization phenotype were caused by amino acid changes in the GP5 protein encoded by ORF5. Site-directed mutagenesis studies unequivocally confirmed that amino acid 98 in the GP5 protein was responsible for the altered neutralization phenotype of these viruses. Amino acid 98 in the GP5 protein, which has not previously been identified as a neutralization determinant of EAV, should be included in an expanded neutralization site D (amino acids 98-106).
KW - Equine arteritis virus
KW - HeLa cells
KW - Neutralization determinant
KW - Persistent infection
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UR - http://www.scopus.com/inward/citedby.url?scp=55749107100&partnerID=8YFLogxK
U2 - 10.1016/j.virusres.2008.09.003
DO - 10.1016/j.virusres.2008.09.003
M3 - Article
C2 - 18851997
AN - SCOPUS:55749107100
VL - 138
SP - 150
EP - 153
JO - Virus Research
JF - Virus Research
SN - 0168-1702
IS - 1-2
ER -