Document Detail


Persistent equine arteritis virus infection in HeLa cells.
MedLine Citation:
PMID:  18579588     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The horse-adapted virulent Bucyrus (VB) strain of equine arteritis virus (EAV) established persistent infection in high-passage-number human cervix cells (HeLa-H cells; passages 170 to 221) but not in low-passage-number human cervix cells (HeLa-L cells; passages 95 to 115) or in several other cell lines that were evaluated. However, virus recovered from the 80th passage of the persistently infected HeLa-H cells (HeLa-H-EAVP80) readily established persistent infection in HeLa-L cells. Comparative sequence analysis of the entire genomes of the VB and HeLa-H-EAVP80 viruses identified 16 amino acid substitutions, including 4 in the replicase (nsp1, nsp2, nsp7, and nsp9) and 12 in the structural proteins (E, GP2, GP3, GP4, and GP5). Reverse genetic studies clearly showed that substitutions in the structural proteins but not the replicase were responsible for the establishment of persistent infection in HeLa-L cells by the HeLa-H-EAVP80 virus. It was further demonstrated that recombinant viruses with substitutions in the minor structural proteins E and GP2 or GP3 and GP4 were unable to establish persistent infection in HeLa-L cells but that recombinant viruses with combined substitutions in the E (Ser53-->Cys and Val55-->Ala), GP2 (Leu15-->Ser, Trp31-->Arg, Val87-->Leu, and Ala112-->Thr), GP3 (Ser115-->Gly and Leu135-->Pro), and GP4 (Tyr4-->His and Ile109-->Phe) proteins or with a single point mutation in the GP5 protein (Pro98-->Leu) were able to establish persistent infection in HeLa-L cells. In summary, an in vitro model of EAV persistence in cell culture was established for the first time. This system can provide a valuable model for studying virus-host cell interactions, especially virus-receptor interactions.
Authors:
Jianqiang Zhang; Peter J Timoney; N James MacLachlan; William H McCollum; Udeni B R Balasuriya
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2008-06-25
Journal Detail:
Title:  Journal of virology     Volume:  82     ISSN:  1098-5514     ISO Abbreviation:  J. Virol.     Publication Date:  2008 Sep 
Date Detail:
Created Date:  2008-08-15     Completed Date:  2008-09-29     Revised Date:  2013-06-05    
Medline Journal Info:
Nlm Unique ID:  0113724     Medline TA:  J Virol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  8456-64     Citation Subset:  IM    
Affiliation:
Department of Veterinary Science, Maxwell H Gluck Equine Research Center, University of Kentucky, Lexington, Kentucky 40546-0099, USA.
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MeSH Terms
Descriptor/Qualifier:
Animals
Antibodies, Monoclonal / metabolism
Arteritis Virus, Equine / classification,  genetics*,  growth & development,  immunology
Arterivirus Infections / veterinary*,  virology
Base Sequence
Carrier State / veterinary*,  virology
Cytogenetic Analysis / veterinary
Electroporation / veterinary
Fluorescent Antibody Technique, Indirect / veterinary
HeLa Cells
Horse Diseases / virology*
Horses
Humans
Molecular Sequence Data
Nucleic Acid Amplification Techniques / veterinary
RNA, Viral / genetics,  isolation & purification
Reverse Transcriptase Polymerase Chain Reaction / veterinary
Sequence Analysis, RNA / veterinary
Viral Nonstructural Proteins / metabolism
Chemical
Reg. No./Substance:
0/Antibodies, Monoclonal; 0/RNA, Viral; 0/Viral Nonstructural Proteins
Comments/Corrections

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